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1. Amlamax in the Management of Dyslipidemia in Humans. Hypercholesterolemia is the major cause of cardiovascular diseases leading to myocardial infractions leading to considerable morbidity and mortality. During the past decade a group of molecules referred to as statins such as simvastatin, atrovastatin have been tried with great success in reducing total cholesterol. These molecules act by inhibiting the HMG CoA reductase enzyme thereby interfering with the synthesis of cholesterol. But statins reduce all the cholesterol including HDL cholesterol. Long term drug vigilance activity has revealed serious side effects of tendinopathy and related musculoskeletal disorders in some of the subjects. In an effort to manage hypercholesterolemia without serious side effects in a natural way we had tried the use of Amlamax a reconstituted, purified, standardized dried extract of amla (Emblica officinalis) containing 30% ellagitannins significant elevation of HDL cholesterol by the administration of Amlamax [Antony B, Merina B, Sheeba, V. Amlamax in the Management of Dyslipidemia in Humans. Indian J Pharm Sci. 2008 Jul-Aug; 70(4): 504-507]
2. A Pilot Clinical Study to Evaluate the Effect of Emblica Officinalis Extract (Amlamax) on Markers of Systemic Inflammation and Dyslipidemia. Emblica officinalis Gaertn, commonly known as the Indian gooseberry of “Amla”, has been used as health food for centuries in India and other Asian countries. The biological effects of amla have been attributed to the antioxidant properties of the low-molecular weight hydrolysable tannins present in the fruit. Amlamax is a purified, standardized, dried extract of amla containing about 35% galloellagi tannins along with other hydrolysable tannins. Our earlier studies on rabbits showed significant reduction in total cholesterol and triglycerides as well as increase in HDL. The present study extends these results to human volunteers. Two doses of the extract were evaluated – 500mg and 1000mg per day for 6 months. Blood samples were collected at the 3rd and 6th months showed reduction in total and LDL cholesterols and enhancement of beneficial HDL cholesterol. In addition, blood CRP levels, a marker for inflammation, were also significantly reduced. Since dyslipidemia and inflammation and the two major components of cardiovascular diseases, the present results must be considered encouraging and indicate the potential of Amlamax in the management of heart disease. [Antony B, Benny M, KaimalTNB. A Pilot Clinical Study to Evaluate the Effect of Emblica Officinalis Extract (Amlamax) on Markers of Systemic Inflammation and Dyslipidemia. Indian Journal of Clinical Biochemistry. 2008; 23(4): 378-381.]
3. Effect of Standardized Amla Extract on Atherosclerosis and Dyslipidemia. Emblica officinalis, commonly known as Indian gooseberry (Amla), is found to be effective for the reversal of dyslipidemia and intima-media thickening and plaque formation in the aorta in hypercholesterolaemic rabbits. In this study, cholesterol powder (100 mg/kg body weight) was administered orally to healthy NZ white rabbits for 4 mo to induce hypercholesterolaemia; and thereafter, amla extract was given in two doses (10 mg and 20 mg/kg/ d orally) for 4 mo. Fasting lipid profile was done monthly and also at the end of treatment. After sacrificing the animals, tissue cholesterol (liver, heart and kidney) and 3-hydroxy-3-methylglutaryl-Coenzyme A reductase activity of liver were estimated and part of aorta and myocardium were processed for histological studies. Feeding of amla extract (10 mg and 20 mg/kg) for 4 mo reversed these changes and the lumen of the aorta became normal as in the normal control group. Reversal of dyslipidemia and atheromatous plaques achieved by amla extract seems to be brought about by a number of factors, such as its ability to prevent low-density lipoprotein oxidation, its antioxidant action, besides decreasing synthesis of cholesterol by inhibiting 3-hydroxy-3-methylglutaryl-Coenzyme A reductase activity and elevating high-density lipoprotein level to enhance reverse cholesterol transport. [Antony B, Merina B, Sheeba V, Mukkadan J. Effect of Standardized Amla Extract on Atherosclerosis and Dyslipidemia. Indian J Pharm Sci. July-August 2006; 68(4): 437-441.]
4. Toxicity Studies of Amlamax – Purified Standardized Extract of Emblica Officinalis. Amlamax is the purified, standardized extract of ‘Amla’ fruits. Acute and subacute toxicity studies were carried out by oral administration of Amlamax at different dosages in mice and rats. Doses up to 2g/kg orally for three months did not produce any haematological, biochemical and histological changes in vital organs.[Antony B, Merina B, Sheeba V. Toxicity Studies of Amlamax – Purified Standardized Extract of Emblica Officinalis. Indian J. Nat. Prod. 2007;23(2):14-17.]
5. Hypolipidemic Activity of Phyllanthus Emblica Linn (Amla) & Trigonella Foenum Graecum (Fenugreenk) Combination In Hypercholesterolemic Subjects – A Prospective, Randomised, Parallel, Open-Label, Positive Controlled Study. The present study is aimed at to evaluate the hypolipidemic and antioxidant activity of combination of fenugreek and amla in hyperlipidemic subjects.Twenty eight subjects were selected from outpatient department of medicine with the inclusion criteria of total cholesterol > 220mg/dl and or serum triglyceride >140mg/dl. They were enrolled for a period of 12 weeks after obtaining written informed consent. Subjects were randomly divided into 2 groups. Group I [test drug combination -1000mg Amla + 500mg Fenugreek/day] and Group II [standard drug Atorvastatin 10 mg daily]. Subjects were evaluated every 4 weeks for 12 weeks. During these visits biochemical investigations of lipid profile and antioxidant activity were done. At the end of 12 weeks, there were significant decrease in serum total cholesterol (20-26%), LDL (25-34%), triglyceride (15-30%) and VLDL (15-30%) levels & an increase in the HDL (0-5%) in the test drug group. This combination has shown similar response on lipid profile as statins with lesser adverse effects. Antioxidant activity of the combination is also demonstrated in the trial. [Joseph S, Santhost D, Udupa AL, Gupta S, Ojeh N, Rathnakar UP, et al. Hypolipidemic Activity of Phyllanthus Emblica Linn (Amla) & Trigonella Foenum Graecum (Fenugreek) Combination In Hypercholesterolemic Subjects – A Prospective, Randomised, Parallel, Open-Label, Positive Controlled Study. Asian Journal of Biochemical and Pharmaceutical Research. 2012;1(2):225-230.]

2/27/2024

Published Studies on Curcumin with Turmeric Essential Oil 

Please note: These studies refer to the blend of curcumin and turmeric essential oil.

1. Curcumin as adjuvant therapy to improve remission in myeloma patients: a pilot randomized trial.
   Background: The treatment for ineligible transplant multiple myeloma is melphalan prednisone. Curcumin has an anti-inflammatory and antiangiogenesis in cancer- directed to nuclear factor-kappa B (NF-kB) pathway. Interleukin 6 (IL-6), vascular endothelial growth factor (VEGF), tumor necrosis factor-alpha (TNF-α), C-reactive protein (CRP), and lactate dehydrogenase (LDH) were also involved in the pathogenesis of myeloma. No clinical study has evaluated the efficacy of curcumin in myeloma patients. To evaluate the efficacy of curcumin as adjuvant into melphalan prednisone in myeloma patients
   Methods: 33 myeloma patients at Dr. Kariadi General Hospital, Semarang, Indonesia during 2016-2017 were randomly assigned single-blindedly into MPC (n=17) and control group (n=16). The MPC group was treated with melphalan 4 mg/m2, prednisone 40 mg/m2 for 7 days, and curcumin 8 gram daily for 28 days. The MP control group was treated with melphalan, prednisone, and placebo. The primary endpoint was the overall remission. Pre and post-treatment was examined for NF-κB, VEGF, TNF-α, IL-6, LDH, and CRP levels All data analyses were per protocol.
   Results: There was a significant difference in overall remission between the MPC and MP control groups [75%vs 33.3%, x2=6.89, P=0.009]. A significant decrease of NF-κB, VEGF, TNF-α levels were shown in the MPC group compared with the MP control group. There was a significant decrease in IL-6 levels in a subgroup analysis of the MPC group. TNF-α levels had a significant correlation with remission [OR=1.35; (95%CI=1.03-1.76); P=0.03].
   Conclusion: Curcumin has an efficacy in improving overall remission and decreasing NF-κB, VEGF, TNF-α, and IL-6 levels in myeloma patients. [Santosa D, Suharti C, Riwant I, et al. Curcumin as adjuvant therapy to improve remission in myeloma patients: a pilot randomized clinical trial. Casp J Int Med. 2022;13: 375-384.]
2. Prospective study to evaluate efficacy, safety and tolerability of dietary supplement of curcumin (BCM95) in subjects with Active relapsing MultipleSclerosis treated with subcutaneous Interferon beta 1a 44 mcg TIW (CONTAIN).
   Background: multiple sclerosis (MS) is a complex disease sustained by several pathogenic mechanisms. As such, combination therapy strategies, targeting a range of disease mechanisms, might represent the ideal therapeutic approach. Here we investigated the efficacy of curcumin, a naturally occurring poly-phenolic phytochemical with potent anti-inflammatory and antioxidant properties, in subjects under treatment with IFN β-1a, to test the effects of this combination therapy on clinical and MRI parameters of inflammation and neurodegeneration in relapsing MS (RMS).
   Methods: eighty active RMS were prospectively enrolled, randomized (1:1) to either the IFN-curcumin or the IFN-placebo group and followed up longitudinally with clinical and MRI assessments for 24 months. Primary endpoint was the efficacy of curcumin versus placebo as add-on therapy on new/enlarging T2 lesions in RMS subjects under treatment with subcutaneous IFN β-1a 44 mcg TIW. Efficacy on clinical parameters (relapses and disability progression), other MRI parameters of inflammation (T1 Gd-enhancing lesions, combined unique active-CUA lesions) and neurodegeneration (T1-hypointense lesions, grey matter loss and white matter microstructural damage) as well as safety and tolerability of curcumin were explored as secondary endpoints.
   Results: ten subjects dropped out from the study by month 12 (6 in the IFN-curcumin group and 4 in the IFN-placebo group), and 27 by month 24 (11 in the IFN-curcumin group and 16 in the IFN-placebo group). Although no between-group difference was present in terms of proportion of subjects free from new/enlarging T2 lesions, a lower proportion of patients with CUA lesions was noted at month 12 in the IFN-curcumin group in comparison with the IFN-placebo group (7.5% vs 17.5%, χ² test p= 0.0167). This result was not confirmed at month 24. The statistical analysis failed to reveal any difference between the two treatment groups—IFN-curcumin and IFN-placebo—in terms of relapses, disability progression, other MRI metrics of inflammation and MRI changes suggestive of ongoing neurodegeneration. No difference in the rate and nature of adverse events was observed between the two treatment groups.
   Conclusion: Although the study drop-out rate was too high to allow definite conclusions, our findings suggest that curcumin might add to IFN β-1a efficacy on radiological signs of inflammation in MS, while it did not seem to exert any neuroprotective effect as assessed by clinical and MRI parameters. [Petracca M, Quarantelli M, Moccia am, et al. ProspeCtive study to evaluate efficacy, safety and tOlerability of dietary supplemeNT of curcumin (BCM95) in subjects with Active relapsing MultIpleSclerosis treated with subcutaNeous Interferon beta 1a 44 mcg TIW (CONTAIN). Mult Scler Relat Disord. 2021 Sep 21;56:103274.
3. A randomized, placebo-controlled study to evaluate the effect of bio-enhanced turmeric formulation on radiation-induced oral mucositis.
   Introduction: Oral mucositis is the most common toxicity of chemoradiotherapy treatment of head and neck cancers. The present study was performed to evaluate the effect of a researched turmeric formulation on oral mucositis in patients receiving chemoradiotherapy for oral cancer.
   Methods: This randomized double-blinded placebo-controlled trial included 60 patients with oral cancer who had undergone radical surgery. Patients were equally randomized into 3 arms. Bio- enhanced turmeric formulation (BTF) capsules (low dose [1 g/day] or high dose [1.5 g/day]) or placebo was administered daily for 6 weeks with concurrent chemoradiotherapy. Study endpoints included the impact of the treatment on chemoradiotherapy-induced oral mucositis along with dysphagia, oral pain, dermatitis, and weight loss.
   Results: The incidence of grade 3 toxicity of oral mucositis, oral pain, dysphagia, and dermatitis was significantly lower in patients who received BTF than placebo. Twenty-five and 20% patients in BTF 1 g/day (p = 0.011) and 1.5 g/day (p = 0.004) arms, respectively, developed grade 3 oral mucositis compared to 65% patients in the placebo arm. Thirty-five and 30% patients in BTF 1 g/day (p = 0.027) and 1.5 g/day (p = 0.011) arms, respectively, developed grade 3 oral pain compared to 70% patients in the placebo arm. Twenty-five and 20% patients in BTF 1 g/day (p = 0.025) and 1.5 g/day (p = 0.010) arms, respectively, developed grade 3 dysphagia compared to 60% patients in the placebo arm. Ten and 5% patients in BTF 1 g/day (p = 0.114) and 1.5 g/day (p = 0.037) arms. respectively, developed grade 3 dermatitis compared to 30% patients in the placebo arm. Patients under BTF supplementation experienced significantly less weight loss and greater compliance with treatment than placebo.
   Conclusion: BTF (BCM-95®) can significantly reduce chemoradiotherapy-induced severe oral mucositis, dysphagia, oral pain, and dermatitis in oral cancer patients. [Soni TP, et al. A randomized, placebo-controlled study to evaluate the effect of bio-enhanced turmeric formulation on radiation-induced oral mucositis. ORL J Otorhinolaryngol Relat Spec. 2021 Jun 23;1-11.]
4. Bioavailable turmeric extract for knee osteoarthritis: a randomized, non-inferiority trial versus paracetamol.
   Background: To compare the efficacy and safety of bioavailable turmeric extract versus paracetamol in patients with knee osteoarthritis (OA).
   Methods: In this randomized, non-inferiority, controlled clinical study, patients of knee OA were randomized to receive bioavailable turmeric extract (BCM-95®) 500 mg capsule two times daily or paracetamol 650 mg tablet three times daily for 6 weeks. The primary outcome measure was Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) pain subscale. The secondary outcome measures were WOMAC total, WOMAC stiffness, and WOMAC physical function scores. Responder analysis of individual patients at different levels (≥ 20%, ≥ 50%, and ≥ 70%) for WOMAC score was calculated. TNF alpha and CRP levels were evaluated and adverse events (AE) were also recorded.
   Results: Seventy-one and seventy-three knee OA patients, respectively in bioavailable turmeric extract and paracetamol groups, completed the study. Non-inferiority (equivalence) test showed that WOMAC scores were equivalent in both the groups (p value < 0.05) in all the domains within the equivalence limit defined by effect size (Cohen’s d) of 0.5 whereas CRP and TNF- α were better reduced with turmeric extract than paracetamol. After 6 weeks of treatment, WOMAC total score, pain, stiffness, and function scores got a significant improvement of 23.59, 32.09, 28.5, and 20.25% respectively with turmeric extract. In the turmeric extract group, 18% of patients got more than 50% improvement and 3% of patients got more than 70% improvement in WOMAC pain and function/stiffness score and none of the patients in the paracetamol group met the criteria. CRP and TNF-α got significantly reduced (37.21 and 74.81% respectively) in the turmeric extract group. Adverse events reported were mild and comparatively less in the turmeric extract group (5.48%) than in the paracetamol group (12.68%).
   Conclusion: The results of the study suggest that bioavailable turmeric extract is as effective as paracetamol in reducing pain and other symptoms of knee osteoarthritis and found to be safe and more effective in reducing CRP and TNF-α. [Singhal S, Hasan N, Nirmal K, et al. Bioavailable turmeric extract for knee osteoarthritis: a randomized, non- inferiority trial versus paracetamol. Trials. 2021;22:105.]
5. Biocurcumin as Radiosensitiser for Cervical Cancer Study (BRACES): A Double-Blind Randomised Placebo-Controlled Trial. Cervical cancer is a leading cause of death among women worldwide, particularly in Indonesia. The main treatment of advanced- stage cervical cancer is radiation; however, the outcomes do not meet the required expectations. [1,7-Bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5dione] has been reported in several studies for its potency in cancer therapy. This study aims to investigate the clinical and molecular [(malondialdehyde (MDA) and NF-κB levels] effects, apoptotic index, and safety of Biocurcumin (BCM-95) as a radiosensitiser in cervical cancer. In this double-blind placebo randomised-controlled trial, we randomised 121 patients into 2 groups (BCM-95 or placebo). MDA and their NF-κB levels and apoptotic index were measured before and after administering 24 Gy of radiation. MDA was identified using Wills’ method, whereas NF-κB was identified via ELISA. The apoptotic index was identified using TUNEL and DAPI staining. The clinical response was classified based on the RECIST. MDA levels before radiation were similar between both groups in per protocol and intention-to-treat (ITT) analyses (p = 0.53 and p = 0.16, respectively). After radiation, MDA levels increased in both groups with no significant differences in per protocol and ITT analyses (p = 0.52 and p = 0.18, respectively). NF-κB levels before radiation were similar between the two groups in per protocol and ITT analyses (p = 0.92 and p = 0.98, respectively). After radiation, the BCM-95 group showed an increase in the NF-κB levels compared with the placebo group in per protocol analysis but not in ITT analysis (p = 0.018 and p = 0.42, respectively). The BCM-95 group had a higher apoptotic index before radiation in per protocol analysis but not in ITT analysis (p = 0.01 and p = 0.61, respectively). After radiation, the apoptotic index remained higher in the BCM-95 group in per protocol analysis but not in ITT analysis (p = 0.04 and p = 0.91, respectively). There was no significant difference in complete response between the groups (per protocol, p = 0.61; ITT analysis, p = 0.90). Although BCM-95 can regulate ROS, NF-κB, and apoptosis in human cervical cancer, it is not significant [for radiosensitizer]. Therefore, BCM-95 does not improve clinical response to radiation treatment. [Purbadi S, Rustamadji P, Prijanti AR, et al. Biocurcumin as radiosensitiser for cervical cancer study (BRACES): a double-blind randomised placebo-controlled trial. Evid Based Complement Alternat Med. 2020;2020:1986793.]
6. Role of turmeric extract in minimising mucositis in patients receiving radiotherapy for head and neck squamous cell cancer: a randomised, placebo-controlled trial.
   Objective: To determine the role of turmeric extract in reducing mucositis in patients undergoing radiotherapy for head and neck cancer.
   Methods: Sixty-one patients who underwent radiotherapy were included in the study and randomised into groups A and B. Patients in group A received 500 mg of turmeric extract (BCM-95) thrice daily, while patients in group B received placebo until radiotherapy completion. All patients were assessed for oral mucositis on a weekly basis during treatment and two months post- treatment using the National Cancer Institute Common Terminology Criteria for Adverse Events and World Health Organization criteria.
   Results: Both groups had a similar grade of mucositis in first two weeks of treatment. The severity of mucositis was progressive in the control group, with four patients developing grade 3 mucositis by week four. In group A, however, the majority of patients (73.3 per cent) had grade 1 mucositis after four weeks of treatment. The difference was statistically significant from the third week onwards (p < 0.001).
   Conclusion: Turmeric extract reduces the incidence and severity of radiation-induced mucositis, which can benefit patients undergoing radiation for head and neck cancer. [Arun P, Sagayaraj A, Azeem Mohiyuddin SM, Santosh D. Role of turmeric extract in minimising mucositis in patients receiving radiotherapy for head and neck squamous cell cancer: a randomised, placebo-controlled trial. J Laryngol Otol. 2020:1-6.]
7. Curcumin and inflammation in non-alcoholic fatty liver disease: a randomized, placebo controlled clinical trial.
   Background: The aim of the present study was to evaluate the effects of curcumin supplementation on inflammatory indices, and hepatic features in patients with non-alcoholic fatty liver disease (NAFLD).
   Methods: Fifty patients with NAFLD were randomized to receive lifestyle modification advice plus either 1500 mg curcumin or the same amount of placebo for 12 weeks.
   Results: Curcumin supplementation was associated with significant decrease in hepatic fibrosis (p < 0.001), and nuclear factor-kappa B activity (p < 0.05) as compared with the baseline. Hepatic steatosis and serum level of liver enzymes, and tumor necrosis-α (TNF-α) significantly reduced in both groups (p < 0.05). None of the changes were significantly different between two groups.
   Conclusion: Our results indicated that curcumin supplementation plus lifestyle modification is not superior to lifestyle modification alone in amelioration of inflammation. [Saadati S, et al. Curcumin and inflammation in non-alcoholic fatty liver disease: a randomized, placebo controlled clinical trial. BMC Gastroenterology. 2019 Jul 25;19(1):133.]
8. The effect of curcumin supplementation anthropometric indices, insulin resistance and oxidative stress in patients with type 2 diabetes: a randomized, double-blind clinical trial.
   Background: Diabetes mellitus is a common metabolic disorders in human and affect a lot of people around the world. Curcumin is a component of turmeric and in many studies therapeutic effects such as anti-hypertensive, anti-hyperlipidemia, anti-hyperglycemia for this substance are shown. Aim: The aim of this study was to investigate the effect of curcumin supplementation on anthropometric indices glycemic control and oxidative stress in overweight patients with type 2 diabetes.
   Materials and methods: In this randomized, double- blind, placebo-controlled trial, 53 participants with type 2 diabetes were divided randomly into the experimental and control groups to receive either 1500 mg curcumin or placebo capsule three times in a day for 10 weeks.
   Results: Supplementation with curcumin in type 2 diabetes compare to placebo causes a significant changes in mean weight (− 0.64 ± 0.22 vs. 0.19 ± 0.37 p < 0.05), body mass index (BMI) (0.3 ± 0.03 vs. 0.1 ± 0 p < 0.05), waist circumference (WC) (− 1.2 ± 0.4 vs. − 0.43 ± 0.11 p < 0.05) and fasting blood sugar (FBS)
(− 7 ± 2 vs. 3 ± 0.2 p < 0.05) but did not show any difference for hemoglobin A1c (HbA1c), insulin, malondialdehyde (MDA), total antioxidant capacity (TAC), Homeostatic Model Assessment for Insulin Resistance (HOMA-IR) and pancreatic B cell function (HOMA-B) at end of study.
   Conclusion: This study indicated that daily administration of 1500 mg curcumin has positive effects in reducing fasting blood glucose and weight in patients with type 2 diabetes. [Hodaei H, Adibian M, Nikpayam O, Hedayati M, Sohrab G. The effect of curcumin supplementation anthropometric indices, insulin resistance and oxidative stress in patients with type 2 diabetes: a randomized, double-blind clinical trial. Diabetol Metab Syndr. 2019 May 27;11:41.]
9. Potential therapeutic effects of Curcuma longa extract in patients with osteoarthritis: a randomized, controlled trial.
   Background: Osteoarthritis (OA) is the most common degenerative joint disorder in the elderly and a major public health problem in worldwide. Non-steroidal anti-inflammatory drug (NSAID) is a common medication given in OA patients, but its use was limited due to many side effects. Previous studies showed that Curcuma Longa extracts may exhibit benefic effects in the treatment of OA. To determine the effective and safety of Curcuma Longa extracts for reducting pain in patients with osteoarthritis.
   Methods: A randomized controlled trial (RCT) in OA patients. Subjects were randomized to 3 different group. Group I: CB extract (350 mg of Curcuma longa and 150 mg Boswellia serrata) and NSAID (400 mg ibuprofen or 50 mg diclofenac sodium), group II: CB extract, group III: NSAID. Each subject would be followed up 3 times: baseline, second weeks, fourth weeks after baseline. The pain severity was measured using visual analogue scale (VAS). The analysis is intention to treat based.
   Results: There were 105 subjects enrolled at the study. Subjects were dominated by female (80%) with mean aged 63 years. Ninety-five subjects (group I: 36; group II: 29, group III: 30) remained for complete analysis. Group I showed the greatest reduction of VAS score after the second and fourth weeks of treatment (more than 50%). Group III has the least VAS score reduction after fourth weeks from baseline. There was statistically different of VAS score reduction between groups (P<0.001). The most frequent AE were reported from subjects in group III.
   Conclusion: CB extract is effective and safe for reducing pain in OA patients. [Pinzon RT, Buwana F. J Pharm Sci Res. 2019;11:3628-3623.]
10. The effect of curcumin on high-sensitivity C-reactive protein, serum adiponectin, and lipid profiles in type 2 diabetes. Diabetes mellitus is one of the most common and important metabolic diseases in human. Curcumin, which is a natural polyphenol found in turmeric, can be used in treatment of diabetes complications for its antidiabetic, anti-inflammatory, and antioxidant properties. In this double-blind randomized clinical trial, 44 patients with Type 2 diabetes randomly assigned to curcumin or placebo group. Patients consumed either 1,500- mg curcumin or placebo daily for 10 weeks. Anthropometric measurements were measured at baseline and at the end of the study. Serum concentrations of triglyceride (TG), total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, high- sensitivity C-reactive protein, and adiponectin were determined after 12-hr fasting at the beginning and end of study. The mean serum level of TG decreased in curcumin group compared with baseline (109 ± 36 vs. 124 ± 36; p < 0.05). At the end of study, the mean concentration of high-sensitivity C-reactive protein decreased in the curcumin group compared to the control (2.9 ± 2.9 vs. 3.4 ± 4.2; p < 0.05). The mean serum concentration of adiponectin increased (64 ± 3 vs. 63 ± 4; p < 0.05) in the treatment group compared with the placebo at the end of the study. The results of the current study indicate that curcumin consumption may reduce diabetes complications through decreasing TG level as well as indicators of inflammation. [Adibian M, Hodaei H, Nikpayam O, Sohrab G, Hekmatdoost A, Hedayati M. The effects of curcumin supplementation on high-sensitivity C-reactive protein, serum adiponectin, and lipid profile in patients with type 2 diabetes: A randomized, double- blind, placebo-controlled trial. Phytother Res. 2019 May;33(5):1374-1383.]
11. Safety and efficacy of curcumin versus diclofenac in knee osteoarthritis: a randomized open-label parallel-arm study.
    Background: The purpose of this study was to compare the efficacy and safety of curcumin with those of diclofenac in the treatment of knee osteoarthritis (OA).
    Methods: In this randomized, open-label, parallel, active controlled clinical study, 139 patients with knee OA were randomly assigned to receive either a curcumin 500-mg (BCM-95®) capsule three times daily or a diclofenac 50-mg tablet two times daily for 28 days. Patients underwent assessment at baseline and days 7, 14, and 28. The main outcome measure was severity of pain using visual analogue scale score at days 14 and 28. Knee Injury and Osteoarthritis Outcome Score (KOOS) (at days 14 and 28), anti- flatulent effect (at day 7), anti-ulcer effect, weight-lowering effect, and patient’s and physician’s global assessment of therapy at day 28 were included as secondary outcome measures. Safety after treatment was evaluated by recording adverse events and laboratory investigation. Results: At days 14 and 28, patients receiving curcumin showed similar improvement in severity of pain and KOOS scale when compared with diclofenac, and the difference was not statistically significant. At day 7, the patients who received curcumin experienced a significantly greater reduction in the number of episodes of flatulence compared with diclofenac (P <0.01). At day 28, a weight-lowering effect (P <0.01) and anti- ulcer effect (P <0.01) of curcumin were observed. None of the patients required H2 blockers in the curcumin group, and 19 patients required H2 blockers in the diclofenac group (0% versus 28%, respectively; P <0.01). Adverse effects were significantly less in the curcumin group (13% versus 38% in the diclofenac group; P <0.01). Patient’s and physician’s global assessment of therapy was similar in the two treatment groups.
    Conclusion: Curcumin has similar efficacy to diclofenac but demonstrated better tolerance among patients with knee OA. Curcumin can be an alternative treatment option in the patients with knee OA who are intolerant to the side effects of non-steroidal anti-inflammatory drugs. [Shep D, Khanwelkar C, Gade P, Karad S. Safety and efficacy of curcumin versus diclofenac in knee osteoarthritis: a randomized open-label parallel-arm study. Trials. 2019 Apr 11;20(1):214.]
12. Role of curcumin as an adjuvant in treatment of advanced head and neck squamous cell carcinoma.
    Background: Chemoradiation forms the major line of treatment in advanced head and neck squamous cell carcinoma, but the benefit of chemotherapeutic agents is at the expense of various toxicities. Curcumin has demonstrated promising results in in-vivo and in-vitro studies as a radiosensitiser. The objective of the study was to determine the role of curcumin as an adjuvant in patients undergoing chemo radiation for advanced head and neck cancers.
    Methods: Study involved 21 patients who underwent chemo radiotherapy for advanced head and neck cancers. They were randomized into two groups. Group A received 500 mg of curcumin while, Group B received placebo along with chemoradiation. The response was assessed using RECIST criteria at three months post treatment using contrast enhanced computerized tomography scan. Results: Overall 58.3% patients had partial response and 41.7% patients had stable disease in group A. In group B, 33.3% patients had a partial response and 66.6% patient had a stable disease. Conclusions: Patients receiving curcumin along with chemoradiation had a marginal decrease in tumour volume and 58.3% patients had partial response and 41.7% had stable disease. A statistical significance could not be achieved due to lack of stage-match controls. Further studies are required to validate the role of curcumin as an adjuvant in the treatment of head and neck squamous cell carcinomas. [Arun P, Sagayaraj A, Azeem Mohiyuddin SM, Santhosh D. Role of curcumin as an adjuvant in treatment of advanced head and neck squamous cell carcinoma. Int J Otorhinolaryngol Head Neck Surg. 2018 Nov;4(6):1388-1393.]
13. Effect of Biocurcumax™ Curcumin (BCM-95) On Treatment of Moderate Chronic Periodontitis.
    Background and purpose: Controlling inflammation is a major approach in periodontal treatments, but scaling and root planing are not always effective enough. Curcumin is anti-inflammatory and can adjust inflammatory reactions and its efficacy and immunity is proven. The present research aimed at evaluating the potential of Curcumin BCM-95 in treatment of patients with chronic periodontitis.
    Materials and methods: In a double blind clinical trial, the clinical parameters including, Gingival Sulcus Bleeding Index (GSBI), Loe and Silness Gingival Index (GI), Probing Pocket Depth (PPD), and Clinical Attachment Level (CAL) were recorded at the beginning of the study, at week 6, and month 4. In case group, patients with moderate chronic periodontitis who had no systemic disease with at least one periodontal pocket with 4-6mm depth in each quadrant and bleeding on probing were chosen. After scaling and root planing, the patients took 2 Curcumin oral capsules per day for 4 weeks. The patients in the control group were given placebos.
    Results: The effect of time was found to be significant in PPD, GI, CAL, and GSBI. Moreover, significant differences were seen between PPD average measurements before medication, at first follow up, and second follow-up (P<0.05). But, in GI, GSBI, and CAL the group effect was not significant. In other words, the reduction was seen in these parameters in both groups but they were not significant.
    Conclusion: The effect of Biocurcumax TM Curcumin (BCM-95) was significant in treatment of moderate chronic periodontitis in PPD between the two groups which reduced this parameter. [Amoian B, Ehsani H, Moghadamnia A, Satari FD, Ehsani H. Effect of Biocurcumax™ Curcumin (BCM-95) On Treatment of Moderate Chronic Periodontitis. J Mazandaran Univ Med Sci. 2018;27(158):45-55.]
14. A Randomized Double-Blind Placebo-Controlled Phase IIB Trial of Curcumin in Oral Leukoplakia. Oral leukoplakia is a potentially malignant lesion of the oral cavity, for which no effective treatment is available. We investigated the effectiveness of curcumin, a potent inhibitor of NF-kB/COX-2, molecules perturbed in oral carcinogenesis, to treat leukoplakia. Subjects with oral leukoplakia (n = 223) were randomized (1:1 ratio) to receive orally, either 3.6 g/day of curcumin (n = 111) or placebo (n = 112), for 6 months. The primary endpoint was clinical response obtained by bi-dimensional measurement of leukoplakia size at recruitment and 6 months. Histologic response, combined clinical and histologic response, durability and effect of long-term therapy for an additional six months in partial responders, safety and compliance were the secondary endpoints. Clinical response was observed in 75 (67.5%) subjects [95% confidence interval (CI), 58.4–75.6] in the curcumin and 62 (55.3%; 95% CI, 46.1–64.2) in placebo arm (P = 0.03). This response was durable, with 16 of the 18 (88.9%; 95% CI, 67.2–96.9) subjects with complete response in curcumin and 7 of 8 subjects (87.5%) in placebo arm, demonstrating no relapse after 6 months followup. Difference in histologic response between curcumin and placebo was not significant (HR, 0.88, 95% CI, 0.45–1.71; P = 0.71). Combined clinical and histologic response assessment indicated a significantly better response with curcumin (HR, 0.50; 95% CI, 0.27–0.92; P = 0.02). Continued therapy, in subjects with partial response at 6 months, did not yield additional benefit. The treatment did not raise any safety concerns. Treatment of oral leukoplakia with curcumin (3.6 g for six months), thus was well tolerated and demonstrated significant and durable clinical response for 6 months. [Kuriakose MA, et al. A Randomized Double-Blind Placebo-Controlled Phase IIB Trial of Curcumin in Oral Leukoplakia. Cancer Prevention Research. 2016;9:693-691.]
15. Curcumin and cognition: a randomised, placebo-controlled, double-blind study of community-dwelling older adults. Curcumin therapy in animals has produced positive cognitive and behavioural outcomes; results of human trials, however, have been inconsistent. In this study, we report the results of a 12-month, randomised, placebo- controlled, double-blind study that investigated the ability of a curcumin formulation to prevent cognitive decline in a population of community-dwelling older adults. Individuals (n 96) ingested either placebo or 1500 mg/d BiocurcumaxTM for 12 months. A battery of clinical and cognitive measures was administered at baseline and at the 6-month and 12- month follow-up assessments. A significant time×treatment group interaction was observed for the Montreal Cognitive Assessment (repeated-measures analysis; time×treatment; F=3·85, P<0·05). Subsequent analysis revealed that this association was driven by a decline in function of the placebo group at 6 months that was not observed in the curcumin treatment group. No differences were observed between the groups for all other clinical and cognitive measures. Our findings suggest that further longitudinal assessment is required to investigate changes in cognitive outcome measures, ideally in conjunction with biological markers of neurodegeneration. [Rainey-Smith SR, Brown BM, Sohrabi HR, Shah T, Goozee KG, Gupta VB, Martins RN. Curcumin and cognition: a randomised, placebo-controlled, double-blind study of community-dwelling older adults. Br J Nutr. 2016 Apr 22:1-8 (Epub ahead of print.)]
16. Effect of Curcumin Supplementation During Radiotherapy on Oxidative Status of Patients with Prostate Cancer: A Double Blinded, Randomized, Placebo-Controlled Study. Curcumin is an antioxidant agent with both radiosensitizing and radioprotective properties. The aim of the present study was to evaluate the effect of curcumin supplementation on oxidative status of patients with prostate cancer who undergo radiotherapy. Forty patients treated with radiotherapy for prostate cancer were randomized to the curcumin (CG, n = 20) or placebo group (PG, n = 20). They received curcumin (total 3g/day) or placebo during external-beam radiation therapy of up to 74 Gy. Plasma total antioxidant capacity (TAC) and activity of superoxide dismutase (SOD), catalase, and glutathione peroxidase (GPx) were measured at baseline and 3 mo after radiotherapy completion. Analysis of covariance was used to compare the variables between groups following the intervention. Serum PSA levels and MRI/MRS images were investigated. In CG, TAC significantly increased (P < 0.001) and the activity of SOD decreased (P = 0.018) after radiotherapy compared with those at baseline. In CG, however, the activity of SOD had a significant reduction (P = 0.026) and TAC had a significant increase (P = 0.014) compared with those in PG. PSA levels were reduced to below 0.2 ng/ml in both groups, 3 mo after treatment, however, no significant differences were observed between the 2 groups regarding treatment outcomes. [Hejazi J, Rastmanesh R, Taleban FA, Molana SH, Hejazi E, Ehtejab G and Hara N. Effect of Curcumin Supplementation During Radiotherapy on Oxidative Status of Patients with Prostate Cancer: A Double Blinded, Randomized, Placebo-Controlled Study. Nutrition and Cancer. 2016;0(0):1-9.]
17. Impact of a 3-weeks randomized double-blind cross-over study curcuminoid supplementation on endotoxemia, inflammatory markers, and lipid profiles in healthy overweight and obese adults. Postprandial endotoxaemia (increased bacterial lipopolysaccharide [LPS] level in the circulation) is associated to the increase of pro- inflammatory markers after intake of high-fat high-calorie meals. Endotoxaemia is a potential driver for chronic low-grade inflammation, linked to non-communicable chronic disease. Curcumin (turmeric) has potential anti-inflammatory and hypolipidemic properties; and was shown to attenuate the effect of LPS-induced endotoxaemia in rats. The aim of this study was to investigate whether curcuminoid supplementation affected postprandial endotoxaemia, inflammatory markers, and lipid profiles in humans. Healthy volunteers (n = 16, 50% men and 50% women, aged 19-43 y, BMI 25-44 kg/m², fat mass 19-53%) participated in a double-blinded, placebo-controlled, cross-over study (4 weeks wash-out period). Participants were randomized to 1 capsule per day, curcuminoids (380 mg) or placebo, for three weeks. Postprandial endotoxaemia was induced by single high-fat high-calorie meal intake (929 kcal, 65 g fat, 63 %E). Blood samples were collected before and after each leg of the study. Endotoxaemia markers (sCD14 & LBP) and inflammatory markers (CRP, TNF-α, IL-6, IL- 1β, and IL-10) were measured with immunoassays; lipids were measure colorimetrically. Two participants dropped-out. There was no change in LBP for either trial leg. Subgroup analysis however indicated a 22% decrease in LBP in volunteers with very high fat mass (n = 5) after leg A (p = 0.02). No differences were seen in CRP level after either leg, with large inter-individual variability (36.4-1028.3 ng/mL). sCD14 decreased after both legs (leg A, p = 0.015; leg B, p = 0.019). There were no effects on TNF-α, IL-6, IL-1β. HDL level (beforehigh-fat meal) was significantly higher after leg A (p = 0.01) with no different after the meal. No other effects were seen on total cholesterol, LDL, and triglycerides. HDL is the main lipoprotein removing LPS from the circulation, transporting LPS to the hepatocytes for clearance. Thus increased HDL level could be of benefit to reduce LPS and inhibit subsequent inflammatory responses. Assessing LPS level in plasma will give a better understanding of this effect. [Nuraiza M, Edwards CA, Combet E. Impact of a 3-weeks randomized double-blind cross-over study curcuminoid supplementation on endotoxemia, inflammatory markers, and lipid profiles in healthy overweight and obese adults. Proceedings of the Nutrition Society. 2016 July;75(OCE3):E160.]
18. Curcumin and Major Depression: A Randomised, Double-blind, Placebo Controlled Trial Investigating the Potential of Peripheral Biomarkers to Predict Treatment Response and Antidepressant Mechanisms of Change. This trial provided partial support for the efficacy of supplementation with a patented curcumin extract (500 mg, twice daily BCM-95 Curcumin) for 8 weeks in reducing depressive symptoms in people with major depressive disorder. In the present paper, a secondary, exploratory analysis of salivary, urinary and blood biomarkers collected during this study was conducted to identify potential antidepressant mechanisms of action of curcumin. Pre and post-intervention samples were provided by 50 participants diagnosed with major depressive disorder, and the Inventory of Depressive Symptomatology self-rated version (IDS-SR30) was used as the primary depression outcome measure. Compared to placebo, 8 weeks of curcumin supplementation was associated with elevations in urinary thromboxane B2 (p <.05), and substance P (p
<001); while placebo supplementation was associated with reductions in aldosterone (p <05) and cortisol (p <05). Higher baseline plasma endothelin-1 (rs=−.587; p <01) and leptin (rs=−.470; p <05) in curcumin-treated individuals was associated with greater reductions in IDS-SR30 score after 8 weeks of treatment. Our findings demonstrate that curcumin supplementation influences several biomarkers that may be associated with its antidepressant mechanisms of action. Plasma concentrations of leptin and endothelin-1 seem to have particular relevance to treatment outcome. Further investigations using larger samples sizes are required to elucidate these findings, as the multiple statistical comparisons completed in this study increased the risk of type I errors. [Lopresti AL, Maes M, Maker GL, Hood S, Drummond PD. Curcumin and major depression: A randomised, double-blind, placebo- controlled trial investigating the potential of peripheral biomarkers to predict treatment response and antidepressant mechanisms of change. Eur Neuropsychopharmacol. 2015;25(1):38-50.]
19. Curcumin for the Treatment of Major Depression: A Randomised, Double-blind, Placebo Controlled Study. In this study, 56 individuals with major depressive disorder were treated with BCM-95 curcumin (500 mg twice daily) or placebo for 8 weeks. The primary measure was the Inventory of Depressive Symptomatology self-rated version (IDS-SR30). Secondary outcomes included IDS-SR30 factor scores and Spielberger State-Trait Anxiety Inventory (STAI). From baseline to week 4, both BCM-95 curcumin and placebo were
associated with improvements in IDS-SR30 total score and most secondary outcome measures. From weeks 4 to 8, BCM-95 curcumin was significantly more effective than placebo in improving several mood-related symptoms, demonstrated by a significant group x time interaction for IDS-SR30 total score and IDS-SR30 mood score, and a non-significant trend for STAI trait score. BCM-95 curcumin was shown to have antidepressant effects in people with major depressive disorder, as evidenced by benefits occurring 4 to 8 weeks after treatment. Greater efficacy from curcumin treatment was identified in a subgroup of individuals with atypical depression. [Lopresti AL, Maes M, Maker GL, Hood S, Drummond PD. Curcumin for the treatment of major depression: A randomised, double-blind, placebo controlled study. J Affect Disord. 2014;167:368-375.]
20. A Pilot Clinical Trial of Radioprotective Effects of Curcumin Supplementation in Patients with Prostate Cancer. Patients with prostate cancer receiving radiation therapy usually experience several side effects and these toxicities are sometimes dose limiting. The purpose of this investigation was to assess the radioprotective effects of BCM-95 Curcumin supplementation in patients with prostate cancer. Forty prostate cancer patients undergoing external beam radiotherapy (EBRT) were randomly assigned to curcumin group, taking 3 g/d curcumin (6 × 500 mg capsules of BCM95 n=20), or placebo group (n=20). Analysis of covariance was used to compare radiotherapy related symptoms between groups following the intervention, adjusted for baseline symptoms. The change in urinary symptoms across the 20-week period differed significantly between groups (p=0.011) and patients in the BCM-95 Curcumin group experienced much milder urinary symptoms compared with the placebo group. BCM-95 Curcumin can confer radioprotective effect in patients with prostate cancer who undergo radiation therapy through reducing the severity of radiotherapy related urinary symptoms. [Hejazi J, Rastmanesh R, Taleban F, Molana S, Ehtejab G. A pilot clinical trial of radioprotective effects of curcumin supplementation in patients with prostate cancer. J Cancer Sci Ther. 2013;5:320-324.]
21. Efficacy and Safety of Curcumin in Major Depressive Disorder: A Randomized Controlled Trial. Curcumin, an active ingredient of Curcuma longa Linn (Zingiberaceae), has shown potential antidepressant-like activity in animal studies. The objectives of this trial were to compare the efficacy and safety of curcumin with fluoxetine in patients with major depressive disorder (MDD). Herein, 60 patients diagnosed with MDD were randomized in a 1:1:1 ratio for six weeks observer-masked treatment with fluoxetine (20 mg) and curcumin (1000 mg) individually or their combination. The primary efficacy variable was response rates according to Hamilton Depression Rating Scale, 17-item version (HAM-D17). The secondary efficacy variable was the mean change in HAM-D17 score after six weeks. We observed that curcumin was well tolerated by all the patients. The proportion of responders as measured by the HAM-D17 scale was higher in the combination group (77.8%) than in the fluoxetine (64.7%) and the curcumin (62.5%) groups; however, these data were not statistically significant (P= 0.58). Interestingly, the mean change in HAM-D17 score at the end of six weeks was comparable in all three groups (P= 0.77). This study provides first
clinical evidence that curcumin may be used as an effective and safe modality for treatment in patients with MDD without concurrent suicidal ideation or other psychotic disorders. [Sanmukhani J, Satodia V, Trivedi J, Patel T, Tiwari D, Panchal B, Goel A, Tripathi CB. Efficacy and safety of curcumin in major depressive disorder: a randomized controlled trial. Phytother Res. 2013;28(4):579-85.]
22. A Randomized, Pilot Study to Assess the Efficacy and Safety of Curcumin in Patients with Active Rheumatoid Arthritis. In this study, 45 patients with rheumatoid arthritis were randomized into 3 groups, with patients receiving either BCM-95 curcumin 500 mg twice daily, the prescription drug diclofenac sodium (one brand name is Voltaren®) 50 mg twice daily, or a combination of the two. The results were judged using the clinically validated Disease Activity Score (DAS) 28 and also with the American College of Rheumatology (ACR) criteria and scores for pain and swelling in joints. Patients in all 3 groups improved. The curcumin group showed the greatest improvement, and the endpoint scores were significantly better than the patients in the drug group. Using both interventions concurrently did not show any additional benefit with regards to disease scores. Curcumin was found to be safe with no adverse effects in this study. In the drug group. 14% of the patients withdrew because of adverse effects. [Chandran B, Goel A. A randomized, pilot study to assess the efficacy and safety of curcumin in patients with active rheumatoid arthritis. Phytother Res. 2012 Nov;26(11):1719-25.]
23. Comparative Study of the Efficacy of Curcumin and Turmeric as Chemopreventative Agents in Oral Submucous Fibrosis: A Clinical and Histopathological Evaluation. Oral Submucous Fibrosis (OSMF) is a chronic disease of the oral mucosa. Premalignant lesions form, with a high progression rate to oral cancer. The goal of this study was to determine if BCM-95 curcumin and turmeric essential oil could improve health of the tissue and help prevent conversion to oral cancer. Participants were randomized to 3 groups of 16 people each: Group one received 1 capsule of BCM-95 curcumin, 500 mg curcuminioids, twice daily; group 2 received 12 drops of turmeric essential oil, held in the mouth twice daily then swallowed, for an approximate dosage of 600 mg, and the last group was placebo twice daily. Both BCM-95 curcumin and turmeric essential oil reduced oral discomfort/mouth burning significantly. The study lasted 6 months, and there were significant reductions in disease scores for both group 1 and 2 at each measurement. The authors reported “remarkable improvements after only the first 15 days of use.” After 6 months of use, 7 of the 16 participants in the placebo group were in the advanced disease stage (meaning closer to malignancy) compared to only 1 person in the BCM-95 curcumin group. No serious adverse effects were noted, and the authors called for more and larger trials, as this holds good promise for treatment of OSMF in the future.” [Deepa Das A, Balan A, Sreelatha KT. Comparative study of the efficacy of curcumin and turmeric as chemopreventative agents in oral submucous fibrosis: a clinical and histopathological evaluation. JIAOMR; April-June 2010;22(2):88-92.]
24. Human Clinical Study to Evaluate the Bioavailability of BCM-95®. 15 healthy men and women ages 24-45; 8 assigned to plain curcumin and 7 assigned to BCM-95 curcumin. Results: overall, 7-fold increase over course of 12 hours. BCM-95 peak at 1600 ng/g; plain curcumin peak at ~230 ng/g. BCM-95 curcumin remained above 200 ng/g for 12 hours. Plain curcumin remained above 200 ng/g for less than 2 hours. Two hours after ingestion, BCM-95 levels are 10-fold over plain curcumin. [Benny M, Antony B. Bioavailability of BioCurcumax™ (BCM-095™). Spice India. September, 2006:11-15.]
25. A Pilot Cross-Over Study to Evaluate Human Oral Bioavailability of BCM-95®, A Novel Bioenhanced Preparation of Curcumin. This study compared BCM-95 curcumin’s absorption in human subjects to plain curcumin and also to curcumin enhanced with piperine (black pepper extract) and lecithin. The results showed that BCM-95 curcumin was absorbed 7 times (or 700%) better than plain curcumin, and at one time measure point, showed a blood level 10 times that of plain curcumin. BCM-95 was absorbed 6.3 (or 630%) better than curcumin with piperine and lecithin. [Antony B, et al. A pilot cross-over study to evaluate human oral bioavailability of BCM-95CG (Biocurcumax), a novel bioenhanced preparation of curcumin. Ind J Pharm Sci. 2008 Jul-Aug;70(4):445-9.]
26. Six-Month Randomized Placebo-Controlled, Double-Blind, Pilot Clinical Trial of Curcumin in Patients with Alzheimer’s Disease. 34 participants were randomized to either 1 gram BCM-95® curcumin, 4 grams BCM-95 curcumin, or placebo. All participants were over age 50, and had a diagnosis of probable or possible Alzheimer’s disease based on the National Institute of Neurological and Communicative Disorders and Stroke–Alzheimer Disease and Related Disorders Association diagnostic criteria. Some measures were serum markers of amyloid beta, plasma isoprostanes (a measure of oxidative stress) and antioxidant status. Both 1 gram and 4 grams reduced oxidative stress and improved antioxidant status. There were more adverse effects in the placebo group than in either 1 g or 4 g BCM-95 group. There was a noted increase in serum amyloid beta in both 1 g and 4 g groups, but not placebo. The authors noted this “possibly reflected an ability of curcumin to disaggregate amyloid beta deposits in the brain, releasing the amyloid beta for circulation and disposal.” [Baum L, Lam CW, Cheung SK, et al. Six-month randomized placebo-controlled, double- blind, pilot clinical trial of curcumin in patients with Alzheimer’s Disease. J Clin Psychopharmacol. 2008 Feb;28(1):110-3.]
27. Curcumin Effects on Blood Lipid profile in a 6-month Human Study. No significant cholesterol lowering effects found, though authors speculate curcumin has other cardioprotective physiological effects. [Baum L, Cheung SK, Mok VC, et al. Curcumin effects on blood lipid profile in a 6-month human study. Pharmacol Res. 2007 Dec;56(6):509-14.]
    

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    Clinical Studies Using Curcumin Combinations

28. Curcumin and zinc co-supplementation along with a loss-weight diet can improve lipid profiles in subjects with prediabetes: a multi-arm, parallel-group, randomized, double-blind placebo-controlled phase 2 clinical trial.
    Background: Diabetes is one of the major public health concerns. Prediabetes can increase the risk of developing some non- communicable diseases such as type 2 diabetes. Given the increasing trend of prediabetes, it is critical to control it and prevent its complications. Curcumin is a major bioactive component of turmeric. Zinc is an antioxidant nutrient. The present trial aimed to evaluate the effect of curcumin and zinc co-supplementation along with a loss weight diet on serum lipid profiles in overweight or obese patients with prediabetes.
    Methods: Eighty-four participants were randomized to four groups (curcumin (500 mg/day), zinc (30 mg/day), “curcumin and zinc”, and placebo) for 90 days. Serum total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein (HDL), triglycerides (TG), non-HDL, HDL/LDL ratio, weight, BMI, waist circumstance (WC), hip circumstance (HC), physical activity (PA) and dietary intake were determined pre and post-intervention. This study will be conducted at Yazd Diabetes Research Clinic, Shahid Sadoughi University of Medical Sciences.
    Results: Totally, 82 participants were included in the final analysis. After the adjusted PA effect, changes in serum TG (adjusted p = 0.001), LDL (adjusted p = 0.035), non-HDL (adjusted p = 0.003), HDL/LDL ratio (adjusted p = 0.002), and HDL (adjusted p < 0.0001) revealed a significant difference between the groups. However, the changes in weight (adjusted p = 0.004) and BMI (adjusted p = 0.006) were significant but the changes in dietary intake, PA, WC, and HC were non-significant (adjusted p ≥ 0.05). Despite that there was a significant difference for post-intervention HDL levels (adjusted p = 0.016), other lipid profiles showed no significant difference (adjusted p ≥ 0.05).
    Conclusion: The beneficial effects of “curcumin and zinc” co-supplementation was reported for the changes of some lipid profiles (TG, LDL, HDL, non-HDL, and HDL to LDL ratio), BMI, and weight with no positive effects on TC, dietary intake, PA, WC, and HC. Therefore, it may play a potential role in the prevention of macro and microvascular complications. [Karandish M, Mozaffari-khosravi H, Mohammadi SM, Cheraghian B, Azhdari M. Curcumin and zinc co-supplementation along with a loss- weight diet can improve lipid profiles in subjects with prediabetes: a multi-arm, parallel- group, randomized, double-blind placebo-controlled phase 2 clinical trial. Diabetol Metab Syndr. 2022;14:22.]
29. The effect of curcumin and zinc co-supplementation on glycemic parameters in overweight or obese prediabetic subjects: a phase 2 randomized, placebo-controlled trial with a multi-arm, parallel-group design. Management of prediabetes is a critical step to prevent type-2 diabetes. Curcumin and zinc have been studied as an antioxidant, antiinflammatory, and antidiabetic agents. In this clinical trial, 84 subjects were randomized into curcumin (500 mg), zinc (30 mg), zinc and curcumin, and placebo groups for 90 days. At the baseline and the end of the study, the outcomes (fasting plasma glucose (FPG), 2-hour postprandial glucose (2hpp), HbA1C, insulin, insulin sensitivity (IS), insulin resistance (IR), β-cell function (BCF), weight, body mass index (BMI), dietary intake, and physical activity (PA)) were measured. A hypocaloric diet and PA were recommended for all subjects. In total, 82 subjects completed the study. After the intervention, dietary intake, PA, weight, and BCF% did not show a significant difference among the groups. However, subjects taking only zinc and zinc and curcumin groups experienced decreased BMI compared to the placebo (p = .01 and .007, respectively). The three treated groups had improved FPG (p = .01), 2hpp (p = .003), HbA1C (p = .004), insulin (p = .001), IS% (p = .001), and IR (p < .001) compared to the placebo. Based on these results, zinc and curcumin supplementation exerted a beneficial effect on several key glycemic parameters. [Karandish M, Mozaffari-khosravi H, Mohammadi SM, et al. The effect of curcumin and zinc co-supplementation on glycemic parameters in overweight or obese prediabetic subjects: a phase 2 randomized, placebo- controlled trial with a multi-arm, parallel-group design. Phytother Res. 2021;35(8):4377- 4387.]
30. Efficacy and safety of combination of curcuminoid complex and diclofenac versus diclofenac in knee osteoarthritis.
    Background: To compare the efficacy and safety of combination of curcuminoid complex and diclofenac vs diclofenac alone in the treatment of knee osteoarthritis (OA).
    Methods: In this randomized trial, 140 patients of knee OA received either curcuminoid complex 500mg (BCM-95) with diclofenac 50mg 2 times daily or diclofenac 50 mg alone 2 times daily for 28 days. Patients were assessed at baseline, day 14 and day 28. Primary efficacy measures were Knee injury and OA outcome score (KOOS) subscale at day 14 and day 28. Anti-ulcer effect and patient-physician’s global assessment of therapy at day 28 were included as secondary endpoints. Safety after treatment was evaluated by recording adverse events and laboratory investigations.
    Results: Both treatment groups showed improvement in primary endpoints at each evaluation visit. Patients receiving curcuminoid complex plus diclofenac showed significantly superior improvement in KOOS subscales, viz.pain and quality of life at each study visit (P>.001) when compared to diclofenac. Less number of patients required rescue analgesics in curcuminoid complex plus diclofenac group (3%) compared to diclofenac group (17%). The number of patients who required histamine 2 (H2) blockers was significantly less in curcuminoid complex plus diclofenac group compared to diclofenac group (6% vs 28%, respectively; P<.001). Adverse effects were significantly less in curcumoid complex plus diclofenac group (13% vs 38% in diclofenac group; P<.001). Patient’s and physician’s global assessment of therapy favored curcuminoid complex plus diclofenac.
    Conclusion: Combination of curcuminoid complex and diclofenac showed a greater improvement in pain and functional capacity with better tolerability and could be a better alternative treatment option in symptomatic management of knee OA. [Shep D, Khanwelkar C, Gade P, Karad S. Efficacy and safety of combinations of curcuminoid complex and diclofenac versus diclofenac in knee osteoarthritis:a randomized trial. Medicine 2020 99:16]
31. The benefit of Curcuma longa and Boswellia serrata to improve quality of life in osteoarthritis patients: a randomized controlled trial.
    Background: Quality of life (QoL) can be affected by chronic pain in osteoarthritis (OA). Previous studies showed that combination of Curcuma longa (CL) and Boswellia serrata (BS) extract (CB extract) are promising for alternative treatment for pain in OA. This study aimed to measure the benefit of CB extract to improve QoL in patients with OA.
    Methods: This was a randomized controlled trial (RCT) in OA patients. Subjects were randomized to 3 different group (Group 1: CB extract (350 mg of CL and 150 mg BS) and NSAID (400 mg ibuprofen or 50 mg diclofenac sodium); group 2: CB extract; group 3: NSAID). Each medication was taken two times per day for 4 weeks. QoL measured using 5Q-5D-5L that include five dimensiones in 5Q-5D: mobility, self-care, usual activities, pain/discomfort, anxiety/depression, with 5 levels of severity.
    Results: There were 105 subjects at baseline. After 4 weeks of study, remained 95 subjects. The most common problem in group 1, 2, and 3 was pain (n=23, n=8, n=15 respectively). The improvement of level of severity in group 1 after 4 weeks of treatment was significant in mobility problems (p=0.002), pain/discomfort (p<0.001), and anxiety/depression (p=0.008). A significant improvement was only seen in usual activity problems (p=0.013) in group 2. The decrease of level of severity in usual activity problems (p=0.034), pain/discomfort (p=0.005), and anxiety/depression (p=0.018) in group 3 were also statistically significant.
    Conclusions: CB extract was beneficial to improve QoL in OA patients with a less side effect compared to NSAID. [Pinzon RT, Sanyasi RLR, Pramudita EA, Periska SD. The benefit of Curcuma longa and Boswellia serrata to improve quality of life in osteoarthritis patients: a randomized controlled trial. Int J Res Orthop. 2019;5(6):1005- 1014.]
32. Potential therapeutic effects of Curcuma longa extract in patients with osteoarthritis: a randomized, controlled trial.
    Background: Osteoarthritis (OA) is the most common degenerative joint disorder in the elderly and a major public health problem in worldwide. Non-steroidal anti-inflammatory drug (NSAID) is a common medication given in OA patients, but its use was limited due to many side effects. Previous studies showed that Curcuma Longa extracts may exhibit benefic effects in the treatment of OA. To determine the effective and safety of Curcuma Longa extracts for reducing pain in patients with osteoarthritis.
    Methods: A randomized controlled trial (RCT) in OA patients. Subjects were randomized to 3 different group. Group I: CB extract (350 mg of Curcuma longa and 150 mg Boswellia serrata) and NSAID (400 mg ibuprofen or 50 mg diclofenac sodium), group II: CB extract, group III: NSAID. Each subject would be followed up 3 times: baseline, second weeks, fourth weeks after baseline. The pain severity was measured using visual analogue scale (VAS). The analysis is intention to treat based.
    Results: There were 105 subjects enrolled at the study. Subjects were dominated by female (80%) with mean aged 63 years. Ninety-five subjects (group I: 36; group II: 29, group III: 30) remained for complete analysis. Group I showed the greatest reduction of VAS score after the second and fourth weeks of treatment (more than 50%). Group III has the least VAS score reduction after fourth weeks from baseline. There was statistically different of VAS score reduction between groups (P<0.001). The most frequent AE were reported from subjects in group III.
    Conclusion: CB extract is effective and safe for reducting pain in OA patients. [Pinzon RT, Buwana F. Potential therapeutic effects of Curcuma longa extract in patients with osteoarthritis: a randomized, controlled trial. J Pharm Sci Res. 2019;11:3628-3633.]
33. A randomized, controlled trial of Curcuma longa and Boswellia serrata extract in osteoarthritis.
    Background: Chronic pain is the major complaint in subjects with osteoarthritis (OA). Nonsteroid anti-inflammatory drug (NSAID) is still the drug of choice in Indonesia to treat OA patients. The prolonged consumption of NSAID may lead to many adverse events (AE). Some previous studies showed the extract of Curcuma longa and Boswellia serrata is a promising potential as therapeutic interventions against OA.
    Objective: This study aimed to evaluate the effectiveness and safety of CB extract to relieve symptoms in patients with OA.
    Study Design: This was a randomized controlled trial (RCT) in OA patients. The treatment used in this trial were CB extract (350 mg of Curcuma longa and 150 mg Boswellia serrata) and NSAID (400 mg ibuprofen or 50 mg diclofenac sodium). Subjects were randomized to 3 different groups (Group 1: CB extract and NSAID; group 2: CB extract; group 3: NSAID). Each medication was taken two times per day for four weeks. Paracetamol tablet 500 mg gave to each subject as a rescue medication. [Pinzon RT, Sanyasi RDLR, Pramudita EA, Periska S. A randomized, controlled trial of Curcuma longa and Boswellia serrata extract in osteoarthritis. Global J Med Res (H). 2019;19:51-56.]
34. Efficacy and safety of curcumin and its combination with boswellic acid in osteoarthritis: a comparative, randomized, double-blind, placebo-controlled study.
    Background: The aim of this clinical trial was to assess the efficacy and safety of curcuminoid complex extract from turmeric rhizome with turmeric volatile oil (CuraMed®) and its combination with boswellic acid extract from Indian frankincense root (Curamin® [Swedish formula]) vs placebo for the treatment of 40- to 70-year-old patients with osteoarthritis (OA).
    Methods: The effects of CuraMed® 500-mg capsules (333 mg curcuminoids) and Curamin® 500-mg capsules (350 mg curcuminoids and 150 mg boswellic acid) taken orally three times a day for 12 weeks in 201 patients was investigated in a three- arm, parallel-group, randomized, double-blinded, placebo-controlled trial. Primary outcome efficacy measures included OA physical function performance-based tests, the WOMAC recommended index of joint pain, morning stiffness, limitations of physical function, and the patients’ global assessment of disease severity.
    Results: Favorable effects of both preparations compared to placebo were observed after only 3 months of continuous treatment. A significant effect of Curamin® compared to placebo was observed both in physical performance tests and the WOMAC joint pain index, while superior efficacy of CuraMed vs placebo was observed only in physical performance tests. The effect size compared to placebo was comparable for both treatment groups but was superior in the Curamin® group. The treatments were well tolerated.
    Conclusions: Twelve-week use of curcumin complex or its combination with boswellic acid reduces pain-related symptoms in patients with OA. Curcumin in combination with boswellic acid is more effective. Combining Curcuma longa and Boswellia serrata extracts in Curamin® increases the efficacy of OA treatment presumably due to synergistic effects of curcumin and boswellic acid. [Haroyan A, et al. Efficacy and safety of curcumin and its combination with boswellic acid in osteoarthritis: a comparative, randomized, double-blind, placebo-controlled study. BMC Complement Altern Med. 2018 Jan 9;18(1):7.]
35. Efficacy of curcumin, and a saffron/curcumin combination for the treatment of major depression: A randomised, double-blind, placebo-controlled study.
    Background: Several studies have supported the antidepressant effects of curcumin (from the spice turmeric) and saffron for people with major depressive disorder. However, these studies have been hampered by poor designs, small sample sizes, short treatment duration, and similar intervention dosages. Furthermore, the antidepressant effects of combined curcumin and saffron administration are unknown.
    Methods: In a randomised, double-blind, placebo- controlled study, 123 individuals with major depressive disorder were allocated to one of four treatment conditions, comprising placebo, low-dose curcumin extract (250 mg b.i.d.), high-dose curcumin extract (500 mg b.i.d.), or combined low-dose curcumin extract plus saffron (15 mg b.i.d.) for 12 weeks. The outcome measures were the Inventory of Depressive Symptomatology self-rated version (IDS-SR30) and Spielberger State-Trait Anxiety Inventory (STAI).
    Results: The active drug treatments (combined) were associated with significantly greater improvements in depressive symptoms compared to placebo (p=.031), and superior improvements in STAI-state (p < .001) and STAI-trait scores (p=.001). Active drug treatments also had greater efficacy in people with atypical depression compared to the remainder of patients (response rates of 65% versus 35% respectively, p=.012). No differences were found between the differing doses of curcumin or the curcumin/saffron combination.
    Limitations: Investigations with larger sample sizes are required to examine the efficacy of differing doses of curcumin and saffron/curcumin combination. Its effects in people with atypical depression also require examination in larger scale studies. Conclusions: Active drug treatments comprising differing doses of curcumin and combined curcumin/saffron were effective in reducing depressive and anxiolytic symptoms in people with major depressive disorder. [Lopresti AL, Drummond PD. Efficacy of curcumin, and a saffron/curcumin combination for the treatment of major depression: A randomised, double- blind, placebo-controlled study. Journal of Affective Disorders. 2017;207:188-196.]
36. Effect of Infla-Kine supplementation on gene expression of inflammatory markers in peripheral mononuclear cells and on C-reaction protein in blood.
    Background: Chronic inflammation is a predisposing factor to numerous degenerative diseases including cancer, heart failure and Alzheimer’s disease. Infla-Kine is a natural supplement comprised of a proprietary blend of Lactobacillus fermentum extract, burdock seed (arctigenin), zinc, alpha lipoic acid, papaya enzyme and an enhanced absorption bio-curcumin complex (BCM-95®).
    Methods: Infla-Kine was administered twice daily to 24 health volunteers for 4 weeks. Quantitative RT-PCR was used to assess mRNA transcripts of IL-1b, IL8, IL-6, NF-κB, and TNF-α from peripheral blood mononuclear cells (PBMC). C-reactive protein (CRP) was measured from serum. Additionally, quality of life questionnaires were employed to assess general feeling of well-being. Assessments were made before treatment and at conclusion of treatment (4 weeks).
    Results: As compared to pre-treatment, after 4 weeks, a statistically significant reduction of IL8, IL-6, NF-κB, and TNF-α transcripts was observed in PBMC. Furthermore, reduction of IL-1b transcript and serum CRP was observed but did not reach statistical significance. Quality of life improvements were most prevalent in muscle and joint pains.
    Conclusions: Overall, our data demonstrate that twice daily administration of Infla- Kine for 4 weeks reduces inflammatory markers and quality of life in healthy volunteers. [Mikirova NA, Kesari S, Ichim TE, Riordan NH. Effect of Infla-Kine supplementation on the gene expression of inflammatory markers in peripheral mononuclear cells and on C-reactive protein in blood. J Transl Med. 2017;15(1):213.]
37. The efficacy and safety of a combination of glucosamine hydrochloride, chondroitin sulfate and bio-curcumin with exercise in the treatment of knee osteoarthritis: a randomized, double-blind, placebo-controlled study.
    Background: Knee osteoarthritis (OA) conservative treatment aims to delay cartilage degeneration; chondroprotective agents are a valid approach in this sense. A commercially available dietary supplement, CartiJoint Forte, containing glucosamine hydrochloride (GH), chondroitin sulfate (CS) and Bio-Curcumin BCM-95®, was used in this trial. AIM: The aim of this study was to assess the efficacy and safety of CartiJoint Forte combined with physical therapy in treating subjects with knee OA.
    Design: A multicenter, prospective, randomized, double blind, placebo-controlled clinical trial.
    Setting: Outpatients referred to the Rehabilitation Departments of two University Hospitals. POPULATION: Firty-three patients were randomly assigned to an experimental group (N=26) or a control group (N=27). Experimental subjects received two tablets of CartiJoint Forte each day for 8 weeks, while those in the control group were provided with a placebo. Three subjects dropped out during the course of the study.
    Methods: The two groups both received 20 sessions of physical therapy during the course of the trial. Primary outcome was pain intensity, measured both at motion and at rest, using the Visual Analogue Scale (VAS). A secondary outcome was an assessment of knee function by Western Ontario and McMaster Universities Arthritis Index and Lequesne Index, knee ROM, and two inflammation markers (C-reactive protein and erythrocyte sedimentation rate). Each assessment was carried out at baseline (T0) at 8 weeks (T1) and at 12 weeks (T2).
    Results: VAS at rest was found to be reduced between T0 and T1, as well as between T0 and T2 (F=13.712; P=0.0001), with no differences between groups (F=1.724; P=0.191). VAS at motion revealed a significant “group x time-check” interaction (F=2.491; P=0.032), with increasing effect of time on VAS reduction (F=17.748; P=0.0001). This was most pronounced in the experimental gropu at 8 weeks (F=3.437; P=0.045). The Lequesne Index showed reductions at T1 and T2 compared to T0 (F=9.535; P=0.0001), along with group effect, since the experimental group presented a lower score at T2 (F=7.091; P=0.009). No significant changes were found in the knee ROM and inflammation markers.
    Conclusion: CartiJoint Forte, added to physical therapy, may ameliorate pain and help to improve algofunctional score in knee OA patients. CLINICAL REHABILITATION IMPACT: Treatment of knee OA with curcuminoids plus glycosaminoglycans, added to physical therapy, improves VAS at motion and Lequesne Index scores. [Sterzi S, et al. The efficacy and safety of a combination of glucosamine hydrochloride, chondroitin sulfate and bio-curcumin with exercise in the treatment of knee osteoarthritis: a randomized, double-blind, placebo-controlled study. Eur J of Physical and Rehab Med. June 2016;52(3):321-330.]
38. Short Report of a Preliminary Open Study of Synofit-Containing Bio-Curcumin, Greenlipped Mussel and Blackcurrant Leaf Extract in Arthritis. To evaluate the potential benefit of Synofit—an association of Curcumin, Perna canaliculus green-mussels and blackcurrant leaf extracts, a real life open study was performed among 86 adult out patients suffering from Fibromyalgia (n = 22), low back pain (n = 33) or knee osteoarthritis (n =31) who accepted to take 3 tablets a day during 1 week then 2 capsules of Synofit during 2 months in addition to their conventional therapy (mainly analgesics and anti-inflammatory) and then to report their evaluation of this complementary treatment. Statistical analysis included paired t test and when possible Wilcoxon signed rank test. Accordingly, the intermediate analysis showed that already within 4 weeks of treatment, an improvement quoted as “light” was statistically reported in patients with low back pain and knee osteoarthritis but not among those with fibromyalgia on pain, physical condition, global assessment of a benefit, quality of life but not on joint stiffness (although joint stiffness considered for the whole group was statistically improved). The limited number of patients and time duration of the study and the absence of double blind controlled study do not allow concluding on the efficacy but these preliminary analyses obtained from an intermediate analysis are encouraging for further studies. [Qu J, Melot C, Appelboom T. Short Report of a Preliminary Open Study of Synofit-Containing Bio-Curcumin, Greenlipped Mussel and Blackcurrant Leaf Extract in Arthritis. Open Journal of Rheumatology and Autoimmune Diseases. 2015;5:113-117.]
39. The use of an anti-inflammatory supplement in patients with chronic kidney disease. Chronic kidney disease (CKD) is characterized by a continuous reduction in kidney function, increased inflammation, and reduced antioxidant capacity. The objective of this study was to assess the effects of a herbal supplement on systemic inflammation and antioxidant status in non-dialysis CKD patients. Sixteen patients with CKD (56.0±16.0 yrs, 171.4±11.9 cm, 99.3±20.2 kg) were randomly chosen to receive a herbal supplement composed of Curcuma longa and Boswellia serrata, or placebo. Plasma levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), glutathione peroxidase (GPx), and serum C-reactive protein (CRP) were measured at baseline and 8 weeks. Baseline data demonstrated elevated inflammation and low antioxidant levels. A significant time effect (p=0.03) and time x compliance interaction effect (p=0.04) were observed for IL-6. No significant differences were observed for any other variables. This study demonstrates that mild and moderate CKD is associated with chronic inflammation and low antioxidant activity. Systemic inflammation and impaired antioxidant status may be greater in CKD populations with multiple comorbidities. Curcumin and Boswellia serrata are safe and tolerable and helped to improve the levels of an inflammatory cytokine. [Moreillon JJ, et al. The use of an anti-inflammatory supplement in patients with chronic kidney disease. J Complement Integr Med. 2013 Jul 1;10.]
40. Randomized, Controlled Human Clinical Study to Assess the Efficacy and Safety of BCM-95® & Bospure® Compared to Celecoxib in the Management of Knee Osteoarthritis. Originally presented at the Osteoarthritis Research Symposium Internationale (OARSI) Annual World Congress on Osteoarthritis, September 15-18, 2011. San Diego, CA. 28 subjects with diagnosed osteoarthritis of the knee were randomized to a 500 mg blend BCM-95 curcumin and Bospure® Boswellia twice a day or to the prescription drug celecoxib (one brand name is Celebrex®) 100 mg twice a day. Symptom scoring and clinical evaluation yielded superior results on pain relief and distance walked for the BCM- 95 and Bospure blend compared to celecoxib. BCM-95 and Bospure equaled celecoxib on joint flexibility. No serious adverse effects noted. [Kizhakedath R, Antony B, Benny M, Kuruvilla BT. Clinical evaluation of a herbal product (Rhulief™) in the management of knee osteoarthritis. Abstract 316. Osteoarthritis Cartilage. 2011;19(S1):S145-S146.]
    

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    Animal Studies [17]

41. Antidiabetic effect of bio-enhanced preparation of turmeric in streptozotocin- nicotinamide induced type 2 Wistar rats.
    Background: Poor oral bioavailability of curcumin, the active ingredient in turmeric, has limited its therapeutic use in various diseases including diabetes mellitus (DM). Objectives: The present study was aimed at evaluating and comparing the antidiabetic activity as well as pharmacokinetic profile of two turmeric extracts. Materials and methods: Rats were divided into seven groups (n = 6) including Normal control (NC), Diabetic control (DC), two standard control groups- Glibenclamide (GLIB) 5 mg/kg and Metformin (MET) 500 mg/kg, two bio- enhanced turmeric extract (BTE) treated groups (BTE-30 (30 mg/kg), BTE-60 (60 mg/kg)) and one regular turmeric extract treated (RTE) group RTE-30 (30 mg/kg). Treatment was given orally for 30 days. Streptozotocin (60 mg/kg) and Nicotinamide (110 mg/kg) were administered intraperitoneally to induce diabetes. Fasting blood glucose (FBG), oral glucose tolerance test at 60 min and 120 min (OG1 and OG2) were analysed at baseline and at the end of study on Day 29. FBG, fasting serum insulin, and concentration of curcumin and its derivatives present in pancreas were analysed at the end of study on Day 30. Results: Turmeric extract treated groups showed significant
(p < 0.05) blood glucose lowering effect, when compared with DC group. FBG, OG1 and OG2 readings were found significantly (p < 0.05) higher in RTE-30 treated group when compared with BTE-30 treated groups. Turmeric extracts showed improved beta-cell function, insulin sensitivity and decreased insulin resistance. BTE-30 had more pancreatic bioavailability of curcumin than RTE-30. Conclusion: Turmeric extracts demonstrated an antidiabetic effect in streptozotocin-nicotinamide induced type 2 diabetic Wistar rats. BTE extract was found to be an effective agent as compared to RTE in controlling hyperglycemia. [Sayeli VK, Shenoy AK. Antidiabetic effect of bio- enhanced preparation of turmeric in streptozotocin-nicotinamide induced type 2 Wistar rate. J Ayurveda Integr Med. 2021;12(3):474-479.]
42. Curcugreen [BCM-95] treatment prevented splenomegaly and other peripheral organ abnormalities in 3xTg and 5xFAD mouse models of Alzheimer’s disease. Metabolic dysfunction and immune disorders are common in Alzheimer’s disease (AD). The mechanistic details of these epiphenomena in AD are unclear. Here, we have investigated whether a highly bioavailable curcuminoid formulation, curcugreen (CGR), can prevent abnormalities in peripheral organs of two mouse models of AD. Eighteen- and 24-month-old male and female 3xTg and 5xFAD mice were treated with CGR (100 mg/kg) for 2 months, orally. Cytoarchitectural changes of spleen, liver, kidney and lungs were studied by H&E stain. Apoptotic death was confirmed by TUNEL staining.
Amyloid deposition, pTau levels, proinflammatory, anti-inflammatory and cell death/survival markers were studied by Western blots. Curcugreen reduced the observed splenomegaly (3xTg) and degeneration of spleen, granulomatous inflammation in the kidney, hepatic sinusoidal disorganization, hepatocellular hypertrophy, inflammation of the central hepatic vein, infiltration and swelling of lung tissues, and apoptotic death in all these areas in both 3xTg and 5xFAD mice. Similarly, CGR decreased amyloid deposition, pTau, proinflammatory markers, cell loss and decrements in anti- inflammatory markers in both 3xTg and 5xFAD mice. Peripheral organ abnormalities and inflammatory responses in AD were ameliorated by curcuminoid treatment. [Manna J, Dunbar GL, Maiti P. Curcugreen treatment prevented splenomegaly and other peripheral organ abnormalities in 3xTg and 5xFAD mouse models of Alzheimer’s disease. Antioxidants (Basel). 2021;10(6):899.]
43. Neuroprotective effect of turmeric extract in combination with its essential oil and enhanced brain bioavailability in an animal model. Purpose. The study evaluated the neuroprotective effect and pharmacokinetic profile of turmeric extract and their metabolites in the blood and brain in an aluminum-induced neurotoxic animal model. Methods. Swiss albino mice received turmeric extract (TE), TEessential oil combination (TE+EO) at doses of 25 and 50 mg/kg/day orally, vehicle (control), and a positive control group. Neurotoxicity was induced by injecting aluminum chloride (40 mg/kg/day, i.p.), and the effect of the intervention was studied for 45 days. The pharmacokinetic and behavioral biochemical markers of brain function and brain histopathological changes were evaluated. Results. The AUC 0-t showed a 30.1 and 54.2 times higher free curcumin concentration in plasma with 25 mg/kg and 50 mg/kg of TE+EO vs. TE, respectively. The concentration of free curcumin in the brain was 11.01 and 13.71- fold higher for 25 mg/kg and 50 mg/kg of TE+EO vs. TE, respectively. Aluminum impairs spatial learning and memory, which was significantly reversed with TE+EO by 28.6% (25 mg/kg) and 39.4% (50 mg/kg). In the elevated plus maze test, 44.8% (25 mg/kg) and 67.1% (50 mg/kg) improvements were observed. A significant reduction in aluminum-induced lipid peroxidation was observed. Also, the levels of glutathione, acetylcholinesterase, and catalase were improved with TE+EO. Damage to the hippocampal pyramidal cells was averted with TE+EO. Conclusion. The neuroprotective and antioxidant response confirms the benefits of TE+EO against aluminum-induced neurotoxicity. The presence of free curcumin and its metabolites in the brain and plasma establishes its improved bioavailability and tissue distribution. Therefore, the benefits of TE +EO could be harnessed in neurodegenerative diseases. [Banji D, Banji OJ, Srinivas K. Neuroprotective effect of turmeric extract in combination with its essential oil and enhanced brain bioavailability in an animal model. BioMed Res Int. 2021;(January):1-12.]
44. Natural agents inhibit colon cancer cell proliferation and alter microbial diversity in mice. The current study was undertaken to investigate the effect of differentially formulated polyphenolic compound Essential Turmeric Oil-Curcumin (ETO-Cur), and Tocotrienol-rich fraction (TRF) of vitamin E isomers on colorectal cancer (CRC) cells that produce aggressive tumors. Combinations of ETO-Cur and TRF were used to determine the combinatorial effects of ETO-Cur and TRF-mediated inhibition of growth of CRC cells in vitro and HCT- 116 cells xenograft in SCID mice. 16S rRNA gene sequence profiling was performed to determine the outcome of gut microbial communities in mice feces between control and ETO-Cur-TRF groups. Bacterial identifications were validated by performing SYBR-based Real Time (RT) PCR. For metagenomics analysis to characterize the microbial communities, multiple software/tools were used, including Quantitative Insights into Microbial Ecology (QIIME) processing tool. We found ETO-Cur and TRF to synergize and that the combination of ETO-Cur-TRF significantly inhibited growth of HCT-116 xenografts in SCID mice. This was associated with a marked alteration in microbial communities and increased microbial OTU (operation taxonomic unit) number. The relative abundance of taxa was increased and the level of microbial diversity after 34 days of combinatorial treatment was found to be 44% higher over the control. Shifting of microbial family composition was observed in ETO-Cur-TRF treated mice as evidenced by marked reductions in Bacteroidaceae, Ruminococcaceae, Clostridiales, Firmicutes and Parabacteroids families, compared to controls. Interestingly, during the inhibition of tumor growth in ETO-Cur treated mice, probiotic Lactobacillaceae and Bifidobacteriaceae were increased by 20-fold and 6-fold, respectively. The relative abundance of anti-inflammatory Clostridium XIVa was also increased in ETO-Cur-TRF treated mice when compared with the control. Our data suggest that ETO-Cur-TRF show synergistic effects in inhibiting colorectal cancer cell proliferation in vitro and in mouse xenografts in vivo, and might induce changes in microbial diversity in mice. [Farhana L, Sarkar S, Nangia-Makker P, Yu Y, Khosla P, Levi E, et al. (2020) Natural agents inhibit colon cancer cell proliferation and alter microbial diversity in mice. PLoS ONE 15(3): e0229823.]
45. Effect of curcumin supplementation on serum expression of select cytokines and chemokines in a female rat model of nonalcoholic steatohepatitis.
    OBJECTIVE: We recently reported that curcumin supplementation in a metabolically (i.e., Western diet [WD]) and chemically (i.e., CCl4) induced female rat model of non-alcoholic steatohepatitis (NASH) was associated with lower liver pathology scores and molecular markers of inflammation. This occurred when curcumin was given during induction of disease (preventative arm; 8-week WD with or without curcumin [8WD + C vs. 8WD]) as well as when given after disease development (treatment arm; 12-week WD with or without curcumin during weeks 9-12 [12WD + C vs. 12WD]). Herein, we sought to extend our findings from that study by determining the effects of curcumin supplementation on cytokine/chemokine expression in serum collected from these same rats. RESULTS: 24 cytokines/chemokines were assayed. IL-2 (+ 80%) and IL-13 (+ 83%) were greater with curcumin supplementation in the prevention arm. IL-2 (+ 192%), IL-13 (+ 87%), IL-17A (+ 81%) and fractalkine (+ 121%) were higher while RANTES was lower (- 22%) with curcumin supplementation in the treatment arm (p < 0.05 for all). RANTES concentrations also correlated significantly with hepatic pathology scores of inflammation (r = 0.417, p = 0.008). Select serum cytokines/chemokines were affected with curcumin supplementation in this female rat model of NASH. Moreover, curcumin’s effect(s) on RANTES and its association with liver disease pathogenesis and progression may warrant further investigation. [Pickich MB, et al. Effect of curcumin supplementation on serum expression of select cytokines and chemokines in a female rat model of nonalcoholic steatohepatitis BMC Res Notes. 2019 Aug 9;12(1):496.]
46. Curcumin supplementation mitigates NASH development and progression in female Wistar rats. Curcumin, a naturally occurring plant polyphenolic compound, may have beneficial effects in nonalcoholic steatohepatitis (NASH) development. We examined whether curcumin supplementation could be used in both prevention and treatment of NASH with fibrosis. Female Wistar rats were provided ad libitum access to a “western diet” (WD) high in fat (43% kcal), sucrose (29% kcal), and cholesterol (2% w/v), as well as 15% fructose drinking water. Intraperitoneal CC14 injections (0.5 mL/kg) were also administered at weeks 1, 2, 4, and 6 to accelerate development of a NASH with fibrosis phenotype. Rats were randomized to four groups (n = 9-12/group) and fed ad libitum: (1) WD for 8-weeks (8WD), (2) WD enriched with curcumin for 8-weeks (8WD+C; 0.2% curcumin, BCM-95, DolCas Biotech) to assess prevention, (3) WD for 12-weeks (12WD), (4) WD for 8-weeks followed by 4-weeks WD+C (12WD+C) to assess treatment. Curcumin prevention (8WD vs. 8WD+C) attenuated (P < 0.05) histological liver inflammation, molecular markers of fibrosis (Col1a1 mRNA) and a serum marker of liver injury (AST). Curcumin treatment (12WD vs. 12WD+C) reduced (P < 0.05) hepatocellular inflammation, steatosis, NAFLD Activity Scores, and serum markers of liver injury (AST, ALP). Moreover, curcumin treatment also increased hepatic pACC/ACC, ApoB100, and SOD1 protein, and decreased hepatic FGF-21 levels; whereas, curcumin prevention increased hepatic glutathione levels. Both curcumin prevention and treatment reduced molecular markers of hepatic fibrosis (Col1a1 mRNA) and inflammation (TNF-α, SPP1 mRNA). Curcumin supplementation beneficially altered the NASH phenotype in female Wistar rats, particularly the reversal of hepatocellular inflammation. [Cunningham RP, et al. Curcumin supplementation mitigates NASH development and progression in female Wistar rats. Physiol Rep. 2018 Jul;6(14):e13789]
47. Risperidone-induced metabolic dysfunction is attenuated by Curcuma longa extract administration in mice. Antipsychotics, such as risperidone, increase food intake and induce alteration in glucose and lipid metabolism concomitantly with overweight and body fat increase, these biological abnormalities belong to the metabolic syndrome definition (high visceral adiposity, hypertriglyceridemia, hyperglycemia, low HDL- cholesterol and high blood pressure). Curcumin is a major component of traditional turmeric (Curcuma longa) which has been reported to improve lipid and glucose metabolism and to decrease weight in obese mice. We questioned the potential capacity of curcumin, contained in Curcuma longa extract (Biocurcuma™), to attenuate the risperidone-induced metabolic dysfunction. Two groups of mice were treated once a week, for 22 weeks, with intraperitoneal injection of risperidone (Risperdal) at a dose 12.5 mpk. Two other groups received intraperitoneal injection of the vehicle of Risperdal following the same schedule. Mice of one risperidone-treated groups and of one of vehicle-treated groups were fed a diet with 0.05% Biocurcuma™ (curcumin), while mice of the two other groups received the standard diet. Curcumin limited the capacity of risperidone to reduce spontaneous motricity, but failed to impede risperidone-induced increase in food intake. Curcumin did not reduce the capacity of risperidone to induce weight gain, but decreased visceral adiposity and decreased the risperidone-induced hepatomegaly, but not steatosis. Furthermore, curcumin repressed the capacity of risperidone to induce the hepatic over expression of enzymes involved in lipid metabolism (LXRα, FAS, ACC1, LPL, PPARγ, ACO, SREBP2) and decreased risperidone-induced glucose intolerance and hypertriglyceridemia. Curcumin decreased risperidone-induced increases in serum markers of hepatotoxicity (ALAT, ASAT), as well as of one major hepatic pro-inflammatory transcription factor (NFκB: p105 mRNA and p65 protein). These findings support that nutritional doses of curcumin contained in Curcuma longa extract are able to partially counteract the risperidoneinduced metabolic dysfunction in mice, suggesting that curcumin ought to be tested to reduce the capacity of risperidone to induce the metabolic syndrome in human. [Auger F, et al. Risperidone- induced metabolic dysfunction is attenuated by Curcuma longa extract administration in mice. Metab Brain Dis. Feb 2018;33(1):63-77.]
48. Curcumin and metformin-mediated chemoprevention of oral cancer is associated with inhibition of cancer stem cells. Effective chemoprevention is critical for improving outcomes of oral cancer. As single agents, curcumin and metformin are reported to exhibit chemopreventive properties, in vitro as well as in patients with oral cancer. In this study, the chemopreventive efficacy of this drug combination was tested in a 4-nitro quinoline 1-oxide (4NQO) induced mice oral carcinogenesis model. Molecular analysis revealed a cancer stem cell (CSC) driven oral carcinogenic progression in this model, wherein a progressive increase in the expression of CSC specific markers (CD44 and CD133) was observed from 8th to 25th week, at transcript (40-100-fold) and protein levels (P ≤ 0.0001). Chemopreventive treatment of the animals at 17th week with curcumin and metformin indicated that the combination regimen decreased tumor volume when compared to the control arm (0.69+0.03 vs 6.66+2.4 mm3; P = 0.04) and improved overall survival of the animals (P = 0.03). Assessment of the molecular status showed an overall downregulation of CSC markers in the treatment arms as compared to the untreated control. Further, in vitro assessment of the treatment on the primary cells generated from progressive stages of 4NQO-induced mice tissue showed a concordant and consistent downregulation of the CSC markers following combination treatment (P < 0.05). The treatment also inhibited the migratory and self-renewal properties of these cells; the effect of which was prominent in the cultures of early dysplastic tissue (P < 0.002). Collectively, our observations suggest that the combination of curcumin and metformin may improve chemopreventive efficacy against oral squamous cell carcinoma through a CSC-associated mechanism. [Siddappa G, et al. Curcumin and metformin- mediated chemoprevention of oral cancer is associated with inhibition of cancer stem cells. Molecular Carcinogenesis. 2017 November;56(11):2446-2460.]
49. Curcumin sensitizes pancreatic cancer cells to gemcitabine by attenuating PRC2 subunit EZH2, and the lncRNA PVT1 expression. Development of resistance to chemotherapeutic drugs is a major challenge in the care of patients with pancreatic ductal adenocarcinoma (PDAC). Acquired resistance to chemotherapeutic agents in PDAC has been linked to a subset of cancer cells termed ‘cancerstem cells’ (CSCs). Therefore, an improved understanding of the molecular events underlying the development of pancreatic CSCs is required to identify new therapeutic targets to overcome chemoresistance. Accumulating evidence indicates that curcumin, a phenolic compound extracted from turmeric, can overcome de novo chemoresistance and re-sensitize tumors to various chemotherapeutic agents. However, the underlying mechanisms for curcumin- mediated chemosensitization remain unclear. The Enhancer of Zeste Homolog-2
(EZH2) subunit of Polycomb Repressive Complex 2 (PRC2) was recently identified as a key player regulating drug resistance. EZH2 mediates interaction with several long non- coding RNAs (lncRNAs) to modulate epithelial-mesenchymal transition and
cancer stemness, phenomena commonly associated with drug resistance. Here, we report the re-sensitization of chemoresistant PDAC cells by curcumin through the inhibition of the PRC2-PVT1-c-Myc axis. Using gemcitabine-resistant PDAC cell lines, we found that curcumin sensitized chemoresistant cancer cells by inhibiting the expression of
the PRC2 subunit EZH2 and its related lncRNA PVT1. Curcumin was also found to prevent the formation of spheroids, a hallmark of CSCs, and to down-regulate several self-renewal driving genes. In addition, we confirmed our in vitro findings in a xenograft mouse model where curcumin inhibited gemcitabine-resistant tumor growth. Overall, this study indicates clinical relevance for combining curcumin with chemotherapy to overcome chemoresistance in PDAC. [Yoshida K, Toden S, Ravindranathan P, Han H, Goel A. Curcumin sensitizes pancreatic cancer cells to gemcitabine by attenuating PRC2 subunit EZH2, and the lncRNA PVT1 expression. Carcinogenesis. 2017 Oct 1;38(10):1036-1046.]
50. Essential turmeric oils enhance anti-inflammatory efficacy of curcumin in dextran sulfate sodium-induced colitis. Turmeric has been used as a medicinal herb for thousands of years for treatment of various disorders. Although curcumin is the most studied active constituents of turmeric, accumulating evidence suggests that other components of turmeric have additional anti-inflammatory and anti-tumorigenic properties. Herein, we investigated anti-inflammatory efficacy and associated gene expression alterations of a specific, curcumin preparation containing essential turmeric oils (ETO-curcumin) in comparison to standard curcumin at three specific doses (0, 5, 25 or 50 mg/kg), in an animal model of dextran sodium sulfate (DSS)-induced colitis. The present study showed that both ETO and standard curcumin treatments provided protection against DSS-induced inflammation. However, ETO-curcumin improved disease activity index (DAI) dose-dependently, while the anti-inflammatory efficacy of standard curcumin remained constant, suggesting that ETO-curcumin may provide superior anti-inflammatory efficacy compared to standard curcumin. Gene expression analysis revealed that anti-inflammatory cytokines including IL-10 and IL-11 as well as FOXP3 were upregulated in the colon by ETO-curcumin. Collectively, these findings suggest that the combined treatment of curcumin and essential turmeric oils provides superior protection from DSS-induced colitis than curcumin alone, highlighting the anti- inflammatory potential of turmeric. [Toden S, Theiss AL, Wang X, Goel A. Essential turmeric oils enhance anti-inflammatory efficacy of curcumin in dextran sulfate sodium- induced colitis. Sci Rep. 2017 Apr 11;7(1):814.]
51. Systematic and comprehensive investigation of the toxicity of curcuminoid-essential oil complex: A bioavailable turmeric formulation. Curcumin, the active component present in Curcuma longa of the family Zingiberaceae, has a number of pharmacological effects, including potential anti-inflammatory activity. One of the major limitations of curcumin/turmeric extract is its poor absorption through the gastrointestinal tract. Several approaches have been adopted to increase the bioavailability of curcumin, including loading curcumin into liposomes or nanoparticles, complexation with phospholipids, addition of essential oils and synthesizing structural analogues of curcumin. In the present study, the toxicity and safety of one such bioavailable turmeric formulation, curcuminoid-essential oil complex (CEC), the toxicity profile of which has not been reported, were examined using in vivo and in vitro models, as per the guidelines of the Organisation for Economic Co-operation and Development. Investigations of acute toxicity study were performed in rats and mice, and the results revealed no signs and symptoms or toxicity or mortality in any of the animals at the maximum recommended dose level of 5,000 mg/kg body weight. The repeated administration of CEC for 90 days in Wistar rats at a dose of 1,000 mg/kg body weight did not induce any observable toxic effects, compared with corresponding control animals. Mutagenicity/genotoxicity investigations were also performed using a bacterial reverse mutation assay (Ames test), a mammalian bone marrow chromosome aberration test and a mammalian erythrocyte micronucleus test in mice. CEC was found to be non-mutagenic in all three mutagenic investigations. Consequently, the present study indicated that CEC elicited no toxic effects in animals or in vitro. Therefore, following investigations of acute toxicity, repeated dose toxicity and mutagenicity, CEC was deemed a safe, non-toxic pharmacological formulation. [Aggarwal ML, Chacko KM, Kuruvilla BT. Systematic and comprehensive investigation of the toxicity of curcuminoid-essential oil complex: A bioavailable turmeric formulation. Molecular Medicine Reports. 2016;13:592-604. DOI: 10.3892/mmr.2015.4579.]
52. Anti-inflammatory activity of BCM-95 (bio-enhanced formulation of turmeric with increased bioavailability) compared to Curcumin in Wistar rats.
    Objective: To evaluate anti-inflammatory activity of bioenhanced turmeric formulation (BCM-95) compared to commercial Curcumin formulation (Curcuminoids 95%) in Carrageenan- induced acute inflammatory model. Materials and Methods: Thirty six Wistar rats were divided into six groups-Normal control (2 ml of vehicle), Standard control (Indomethacin 10 mg/kg), 2 doses of BCM 95 (10 and 20 mg/kg) and Curcuminoids 95% (10 and 20 mg/kg). Paw volume was measured using a digital plethysmometer. Vehicle or test drugs were given to rats 30 min before carrageenan administration. Baseline paw volume reading (V0) was noted just prior to administration of 0.1 ml of 1% carrageenan to right hind paw of the rat. Test paw volume readings (Vt) were measured at 30, 60, 120, 180, 240, 300 and 360 min, after carrageenan injection. Oedema expressed as increased paw volume (vt-v0) was noted and percentage inhibition of oedema was calculated for all treatment groups. Statistical analysis: Difference between groups were analyzed with ANOVA followed by Tukey test.
    Results: All treatment groups demonstrated significant (p<0.05) anti-inflammatory activity (oedema suppression) compared to normal control. Anti-inflammatory activity of BCM 95 treated groups were comparable to standard control group except at certain time points, whereas the same activity at all-time points with Curcuminoid 95% treated groups were significantly less than standard control group. Percentage inhibition of paw oedema was maximum with standard control group followed by BCM 95 treated groups followed by Curcuminoid 95% treated groups.
    Conclusion: BCM 95 treated groups showed significant anti-inflammatory activity compared to Curcuminoid 95% treated groups. [Vinaykumar S, et al. Anti-inflammatory activity of BCM-95 (bio-enhanced formulation of turmeric with increase bioavailability) compared to Curcumin in Wistar rats. Pharmacogn. J. July-Aug 2016;8(4):380-383.]
53. Evaluation of Antiepileptic and Memory Retention Activity of Curcumin Per SE and in Combination with Antiepileptic Drugs. Antiepileptic activity of curcumin and its combination with phenytoin and sodium valproate were studied in chronic model (14 days) of Maximal Electroshock Seizure (MES) and Pentylenetetrazole (PTZ) induced seizure respectively. Elevated plus maze test was used to study effect of drugs and/or seizures on memory retention in MES and PTZ groups. Curcumin in both doses did not show any significant effect (P = 0.33) on tonic extension, while curcumin 100 mg/kg significantly (P < 0.01) reduced clonic phase compared to vehicle control. Curcumin in 100 mg/kg dose significantly (P < 0.001) inhibited PTZ induced seizure. Addition of curcumin to sub therapeutic dose of sodium valproate showed synergistic effect. Curcumin did not show any effect on memory retention. Inhibition of PTZ induced seizure by curcumin could be due to effect on γ-amino butyric acid receptor (GABA) pathway and its antioxidant property. Curcumin can be effective in absence seizure alone and as add on with sodium valproate. [Anovadiya AP, Sanmukhani JJ, Vadgama VK, Tripathi CB. Asian J Pharm Clin Res. 2013;6(2):145-148.]
54. Chemoprevention and Treatment Efficacy of Curcumin in Combination with Metformin in an in Vivo Oral Carcinogenesis Model. Evaluation of the efficacy of [BCM-95] Curcumin and Metformin in prevention of oral pre-malignant lesion (PML) progression. The animal model was established using 4-6 weeks C57BI/6 mice (N=60); the mice were divided into control arm (N=10) with plain drinking water and the treatment arm (N=50) which received the cancer causing 4-nitroquinoline-oxide. After 17 weeks, the mice were taken off the carcinogen and divided into 4 groups: arm 1 with plain water, arm 2 with curcumin, arm 3 with the drug metformin, and arm 4 with a combination of both curcumin and metformin. The mice were examined at 17th week as well as at the end of the 25 week study period, and samples collected for molecular analysis. The average tumor volume was reduced in the combination arm (0.693±0.034) and the individual arms (curcumin 2.45; metformin 1.45±0.33) as compared to the 4NQO arm (6.65±2.37). The average number of lesions (malignant tumors) per mouse was also reduced in the combination arm (Avg 0.375) and the curcumin arm (Avg 0.25) as compared to the 4NQO arm (Avg 0.8). The overall survival of the combination arm was better when compared to individual treatment (p= 0.0006). The animals in the water control arm remained healthy. Curcumin reduced tumor formation both on its own and in conjunction with metformin. Conclusion: The clinical results suggest that the combination arm is more efficient in chemoprevention. Further studies using the molecular markers and subsequent functional studies are currently ongoing. [Siddappa G, Ravindra D, Kulsum S, et al. Chemoprevention and Treatment Efficacy of Curcumin in Combination with Metformin in an in Vivo Oral Carcinogenesis Model. 5th International Federation of Head and Neck Oncologic Societies (IFHNOS). July 26th-30th 2014, New York, NY. Also presented at the 13th National Conference of Foundation for Head and Neck Oncology (FHNO). September 27th–29th 2013, JAIPUR.]
55. Evaluation of Antidepressant Like Activity of Curcumin and its Combination with Fluoxetine and Imipramine: an Acute and Chronic Study. In animal model of depression, BCM-95 curcumin is compared to generic fluoxetine (one brand name is Prozac®) and imipramine (one brand name is Tofranil®). BCM-95 curcumin performed as well as either prescription anti-depressant drug on all measures of depression. However, adding BCM-95 curcumin to the prescription drugs did not increase antidepressant effects. [Sanmukhani J, et al. Evaluation of antidepressant like activity of curcumin and its combination with fluoxetine and imipramine: an acute and chronic study. Acta Pol Pharm. 2011 Sep-Oct;68(5):769-75.]
56. Oral Bioavailability of BCM-95® in Dogs. This study looked specifically at bioavailability in dogs for veterinary purposes. Six healthy adult male and female dogs were divided between plain curcumin and BCM-95 curcumin (reported as the veterinary NMXCC-95 designation). No adverse effects reported. The BCM-95 group had approximately 7-fold increase in absorption over plain curcumin over 8 hours and approximately 9-fold increase over plain curcumin when measured for 12 hours. [Antony B, Butchin RK, Griffin DW. Bioavailability of a novel, bioenhanced preparation in dogs. Poster Presentation. 2009 ACVIM Forum/Canadian VMA Convention: June 3-6, 2009; Montréal, Québec, Canada.]
57. Enhancing the Absorption of Curcuminoids. Turmeric (Curcuma longa), one of the familiar spice has got number of medicinal properties such as anti-septic, anti- inflammatory, wound healing, anti-oxidant, anti-tumor etc. These properties of turmeric are attributed to the active principle, curcumin and essential oil present in the rhizome. But it is suggested and proved that only 50 – 60 percent of total curcumin is absorbed by animal system. Studies conducted on albino rats and results described in this paper reveal that 96 – 97 percent absorption of curcuminoids by mixing curcumin and standardized essential oil of turmeric. [Antony B, Benny M, Rao SB. Enhancing the Absorption of Curcuminoids. Spice India. July 2005;23-26.]
    

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    Animal Studies Using Curcumin Combinations [6]

58. The Effect of Exercise and Nutritional Supplementation on Proinflammatory Cytokine Expression in Young Racehorses During Training. The inflammatory response to vigorous exercise ranges from the mild symptoms of delayed-onset muscle soreness to debilitating injuries affecting soft tissue, joint, and bone. Although there is a great deal of information available on the inflammatory response to exercise in human athletes, less information is available regarding the inflammatory response to exercise in young horses undergoing training for racing careers. Here, we assessed the cytokine response to exercise in a group of young Thoroughbred racehorses during their initial training. Because there is interest in nonpharmacologic approaches to control or ameliorate exercise-induced inflammation, we also examined the anti-inflammatory effect of a nutritional supplement [containing BCM-95® curcumin, BosPure® boswellia, coenzyme Q10, glycine proprionyl-L-carnitine HCl, and D-ribose] fed to half of the horses undergoing training. Twenty-five Thoroughbred horses aged 2 years were followed through their initial race training. Peripheral blood samples were collected at various times during the exercise for the quantitation of lactic acid, oxidative stress, and inflammatory cytokine gene expression. There was an intensity-dependent effect of exercise on lactate, malondialdehyde, and proinflammatory cytokine gene expression. Although training itself was associated with an overall reduction in inflammatory markers, horses receiving the supplement exhibited further reductions in their indicators of inflammation. As such, this study provides novel evidence of nutritional supplementation reducing postexercise inflammation. [Horohov DW, Sinatra ST, Chopra RK, Jankowitz S, Betancourt A, Bloomer RJ. The effect of exercise and nutritional supplementation on proinflammatory cytokine expression in young racehorses during training. J Equine Vet Sci. 2012 December;32(12):805-15.]
59. BreastDefend™ prevents breast-to-lung cancer metastases in an orthotopic animal model of triple-negative human breast cancer. We have recently demonstrated that a natural dietary supplement BreastDefend (BD), which contains extracts from medicinal mushrooms (Coriolus versicolor, Ganoderma lucidum, Phellinus linteus), medicinal herbs (Scutellaria barbata, Astragalus membranaceus, Curcuma longa), and purified biologically active nutritional compounds (diindolylmethane and quercetin), inhibits proliferation and metastatic behavior of MDA-MB-231 invasive human breast cancer cells in vitro. In the present study, we evaluated whether BD suppresses growth and breast-to lung cancer metastasis in an orthotopic model of human breast cancer cells implanted in mice. Oral application of BD (100 mg/kg of body weight for 4 weeks) by intragastric gavage did not affect body weight or activity of liver enzymes and did not show any sign of toxicity in liver, spleen, kidney, lung and heart tissues in mice. Moreover, BD significantly decreased the change in tumor volume over time compared to the control group (p=0.002). BD treatment also markedly decreased the incidence of breast-to-lung cancer metastasis from 67% (control) to 20% (BD) (p<0.05) and the number of metastases from 2.8 (0.0, 48.0) in the control group to 0.0 (0.0, 14.2) in the BD treatment group (p<0.05). Finally, anti-metastatic activity of BD in vivo was further confirmed by the downregulation of expression of PLAU (urokinase plasminogen activator, uPA) and CXCR4 (C-X-C chemokine receptor- 4) genes in breast tumors. In conclusion, BD may be considered as a biological therapeutic agent against invasive breast cancers. [Jiang J, et al. BreastDefend™ prevents breast-to-lung cancer metastases in an orthotopic animal model of triple-negative human breast cancer. Oncol Rep. 2012 Oct;28(4):1139-1145.]
60. Evaluation of hepatoprotective activity of combination of Phyllanthus niruri and Curcuma longa extracts in Wistar rats. Hepatoprotective activity of combination of Phyllanthus niruri(PN) and Curcuma longa(CL) extract was evaluated against carbon tetrachloride(CCl4) induced liver damage. Combination of PN+CL extract at a dose of 400mg/kg, orally was coadministered with CCl4 (0.5 mg/kg i.p) to rats for 7 days. On 8th day serum enzyme levels such as AST, ALT, ALP, TB were determined. Thiopentone induced sleeping time is estimated as an indirect index of functionality of liver. Liver tissue was used to estimate antioxidants such as MDA, GST levels and for histopathological assessment. There was a significant increase in serum enzyme levels and duration of thiopentone induced sleep time in CCl4 treated rats. Coadministration of PN+CL extract combination with CCl4 significantly prevented the rise in serum enzyme levels and normalize the duration of thiopentone induced sleep time. Combination of PN+CL produced significant reduction and increase in MDA & GST liver levels respectively. Histological section of liver in animals treated with CCl4 showed centrilobular area of necrosis with derangement in hepatic architecture. PN+CL administration prevented these deleterious changes, histological section of liver in rats treated with PN+CL showed normal hepatic parenchyma. Combination of PN+CL extract showed significant hepatoprotection against CCl4 induced liver damage. [Adiga S, et al. Evaluation of hepatoprotective activity of combination of Phyllanthus niruri and Curcuma longa extracts in Wistar rats. Research Journal of Pharmaceutical, Biological and Chemical Sciences. 2012 July – September; 3(3):1260.
61. ProstaCaid™ inhibits tumor growth in a xenograft model of human prostate cancer. We have recently demonstrated that the dietary supplement ProstaCaid™ (PC) inhibits growth and invasive behavior of PC-3 human prostate cancer cells in vitro. In the present study, we evaluated toxicity and whether PC suppresses growth of prostate cancer in a xenograft model of human prostate cancer cells implanted in mice. Here, we show that an oral administration of PC (100, 200 and 400 mg/kg) did not affect body weight or activity of liver enzymes (ALT, AST) and did not show any sign of toxicity in liver, spleen, kidney, lung and heart tissues in mice. In addition, PC treatment resulted in the inhibition of tumor volumes (1024.6±378.6 vs. 749.3±234.3, P<0.001) in a xenograft model of prostate cancer with human hormone refractory (independent) PC-3 prostate cancer cells. Moreover, qRT-PCR analysis demonstrated significant upregulation of expression of CDKN1A (p21) and inhibition of expression of IGF2, NR2F2 and PLAU (uPA) genes by an oral administration of PC in prostate cancer xenografts. Our study demonstrates that the concentrations of the dietary supplement ProstaCaid tested did not show signs of toxicity, and its oral application has significant anticancer activity in vivo and can be considered as an alternative treatment for prostate cancer patients. [Jiang J, et al. ProstaCaid™ inhibits tumor growth in a xenograft model of human prostate cancer. Int J Oncol. 2012 May;40(5):1339-1344.
62. Effect of Citrus Polyphenol- and Curcumin-supplemented Diet on Inflammatory State in Obese Cats. Veterinary study looking at obesity-induced pro-inflammatory state in cats and impact of BCM-95 on liver and inflammatory markers. Showed safe use in cats, and significant impact on interleukin 2 (IL-2) and reduction of AGP (a1-acid glycoprotein) which shows that curcumin impacts hepatocytes (liver cells) to reduce AGP, illustrating that BCM-95 is helping liver cells to behave more like liver cells in non-obese cats. [Leray V, Freuchet B, Le Bloc’h J, Jeusette I, Torre C, Nguyen P. Effect of citrus polyphenol- and curcumin-supplemented diet on inflammatory state in obese cats. Br J Nutr. 2011 Oct;106 Suppl 1:S198-201.]
63. Effect of a topical curcumin preparation (BIOCURCUMAX) on burn wound healing in rats.
    Background: Curcumin, a naturally occurring o-methoxyphenol derivative, has been shown to possess several biological properties including antioxidant (free radical scavenging activity), induction of detoxification enzymes and provides protection against degenerative diseases. Topical applications of compounds with free radical scavenging properties in patients have shown to improve significantly wound healing and protect tissues from oxidative damage.
    Objectives: To assess the effect of a topical curcumin preparation on healing of partial thickness burn wounds in rats.
    Methods: The rats are randomly divided into four groups, comprising of six rats in each group. Partial thickness burn wounds are created by pouring hot molten wax at 80ºC. Group I acts a control, Group 2 receives the standard silver sulphadiazine cream, Group 3 gets 20% curcumin cream, and Group 4 receives the combination of the dexamethasone and curcumin cream. Parameters observed are epithelialization period and wound contraction.
    Results & Discussion: The percentage of wound contraction was significantly increased in the topical curcumin preparation (20%) and silver sulfadiazine group compared to control group. The mean period of epithelization was significantly reduced in topical curcumin preparation (20%) group and silver sulfadiazine group as compared to the control.
    Conclusion: Topical curcumin preparation is effective in healing burn wound and the effect was comparable to that of standard drug i.e. silver sulfadiazine. [Durgaprasad S, Reetesh R, Hareesh K, Rajput R. Effect of a topical curcumin preparation (BIOCURCUMAX) on burn wound healing in rats. Journal of Pharmaceutical and Biomedical Sciences. 2011;8(23):1-3.]

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    Ex Vivo or Cellular Studies [7]

64. Curcumin inhibits polycomb repressive complex 2 through lncRNA-PVT1 and enhances gemcitabine sensitivity in chemoresistant pancreatic cancer. Pancreatic ductal adenocarcinoma (PDAC) is one of the most aggressive malignancies, and a major cause of PDAC-associated mortality is acquisition of resistance to chemotherapy. Curcumin is a phenolic compound extracted from turmeric, and is known for its potent anti-inflammatory, anti-oxidative and anti-tumorigenic properties. Accumulating evidence suggests that curcumin can overcome de-novo chemoresistance and re-sensitize tumors to various chemotherapeutic drugs. Recently, polycomb repressive complex 2 (PRC2) was reported to be involved in drug resistant through interaction with several long non-coding RNAs (lncRNAs). Our data provide previously unrecognized evidence for curcumin-induced sensitization to gemcitabine, through co-modulation of PRC2 and PVT1 in PDAC. These data highlight the potential adjunctive therapeutic role for curcumin together with conventional chemotherapeutic drugs in patients with PDAC. [Yoshida K, Toden S, Ravindranathan P, Goel A. Curcumin sensitizes pancreatic cancer cells to gemcitabine by attenuating PRC2 subunit EZH2, and the lncRNA PVT1 expression. Carcinogenesis. 2017:1-11.]
65. Curcumin mediates chemosensitization to 5-fluorouracil through miRNA-induced suppression of epithelial-to-mesenchymal transition in chemoresistant colorectal cancer. Resistance to cytotoxic chemotherapy is a major cause of mortality in colorectal cancer (CRC) patients. Chemoresistance has been linked primarily to a subset of cancer
cells undergoing epithelial-mesenchymal transition (EMT). Curcumin, a botanical with anti-tumorigenic properties, has been shown to enhance sensitivity of cancer cells to chemotherapeutic drugs, but the molecular mechanisms underlying this phenomenon remain unclear. Effects of curcumin and 5-fluorouracil (5FU) individually, and in combination, were examined in parental and 5FU resistant (5FUR) cell lines. We performed a series of growth proliferation and apoptosis assays in 2D and 3D cell cultures. Furthermore, we identified and analyzed the expression pattern of a subset of putative EMT-suppressive microRNAs (miRNAs) and their downstream target genes regulated by curcumin. Chemosensitizing effects of curcumin were validated in a xenograft mouse model. Combined treatment with curcumin and 5FU enhanced cellular apoptosis and inhibited proliferation in both parental and 5FUR cells, while 5FU alone was ineffective in 5FUR cells. A group of EMT-suppressive miRNAs were upregulated by curcumin treatment in 5FUR cells. Curcumin suppressed EMT in 5FUR cells by downregulating BMI1, SUZ12 and EZH2 transcripts, key mediators of cancer stemness- related polycomb repressive complex subunits. Using a xenograft and mathematical models we further demonstrated that curcumin sensitized 5FU to suppress tumor growth. We provide novel mechanistic evidence for curcuminmediated sensitization to 5FU- related chemoresistance through suppression of EMT in 5FUR cells via upregulation of EMT-suppressive miRNAs. This study highlights the potential therapeutic usefulness of curcumin as an adjunct in patients with chemoresistant advanced CRC. [Toden S, Okugawa Y, Jascur T, Wodarz D, Komarova NL, Buhrmann C, Shakibaei M, Boland, Goel A. Curcumin mediates chemsensitization to 5-fluorouracil through miRNA-induced suppression of epithelial-to-mesenchymal transition in chemoresistant colorectal cancer. Carcinogenesis. 2015 Feb 4 (Epub ahead of print).]
66. Curcumin inhibits cancer-associated fibroblast-driven prostate cancer invasion through MAOA/mTOR/HIF-1α signaling. Cancer-associated fibroblasts (CAFs) are key determinants in the malignant progression of cancer, supporting tumorigenesis and metastasis. CAFs also mediate epithelial to mesenchymal transition (EMT) in tumor cells and their achievement of stem cell traits. Curcumin has recently been found to possess anticancer activities via its effect on a variety of biological pathways involved in cancer progression. In this study, we found that CAFs could induce prostate cancer cell EMT and invasion through monoamine oxidase A (MAOA)/mammalian target of rapamycin (mTOR)/hypoxia-inducible factor-1α (HIF-1α) signaling pathway, which exploits reactive oxygen species (ROS) to drive a migratory and aggressive phenotype of prostate carcinoma cells. Moreover, CAFs was able to increase CXC chemokine receptor 4 (CXCR4) and interleukin-6 (IL-6) receptor expression in prostate cancer cells. However, curcumin abrogated CAF-induced invasion and EMT, and inhibited ROS production and CXCR4 and IL-6 receptor expression in prostate cancer cells through inhibiting MAOA/mTOR/HIF-1α signaling, thereby supporting the therapeutic effect of curcumin in prostate cancer. [Du Y, Long Q, Zhang L, Shi Y, Liu X, Li X, Guan B, Tian Y, Wang X, Li L, He D. Curcumin inhibits cancer-associated fibroblast-driven prostate cancer invasion through MAOA/mTOR/HIF-1α signaling. International Journal of Oncology. 2015;1899:0-0.]
67. Curcumin potentiates antitumor activity of 5-fluorouracil in a 3D alginate tumor microenvironment of colorectal cancer.
    Background: To overcome the limitations of animal-based experiments, 3D culture models mimicking the tumor microenvironment in vivo are gaining attention. Herein, we investigated an alginate-based 3D scaffold for screening of 5-fluorouracil (5-FU) or/and curcumin on malignancy of colorectal cancer cells (CRC).
    Methods: The potentiation effects of curcumin on 5-FU against proliferation and metastasis of HCT116 cell and its corresponding isogenic 5-FU- chemoresistant cells (HCT116R) were examined in a 3D-alginate tumor model.
    Results: CRC cells encapsulated in alginate were able to proliferate in 3D- colonospheres in a vivo-like phenotype and invaded from alginate. During cultivation of cells in alginate, we could isolate 3 stages of cells, (1) alginate proliferating (2) invasive and (3) adherent cells. Tumor-promoting factors (CXCR4, MMP-9, NF-κB) were significantly increased in the proliferating and invasive compared to the adherent cells, however HCT116R cells overexpressed factors in comparison to the parental HCT116, suggesting an increase in malignancy behavior. In alginate,curcumin potentiated 5-FU- induced decreased capacity for proliferation, invasion and increased more sensitivity to 5- FU of HCT116R compared to the HCT116 cells. IC50 for HCT116 to 5-FU was 8nM, but co-treatment with 5 μM curcumin significantly reduced 5-FU concentrations in HCT116 and HCT116R cells (0.8nM, 0.1nM, respectively) and these effects were accompanied by down-regulation of NF-κB activation and NF-κB-regulated gene products.
    Conclusions: Our results demonstrate that the alginate provides an excellent tumor microenvironment and indicate that curcumin potentiates and chemosensitizes HCT116R cells to 5-FU-based chemotherapy that may be useful for the treatment of CRC and to overcome drug resistance. [Shakibaei M, Kraehe P, Popper B, Shayan P, Goel A, Buhrmann C. Curcumin potentiates antitumor activity of 5-flurouracil in a 3D alginate tumor microenvironment of colorectal cancer. BMC Cancer. 2015 Apr 10;15:250.]
68. BCM-95 Curcumin Improves Efficacy of Chemotherapy 5-Fluorouracil in Chemoresistant Colorectal Cancer. More than 15% of colorectal cancer (CRC) patients are resistant to 5-Fluorouracil (5-FU)-based chemotherapeutic regimens, and tumor recurrence rates can be as high as 50-60%. Cancer stem cells (CSC) are capable of surviving conventional chemotherapies that permit regeneration of original tumors. This study investigated the effectiveness of 5-FU and BCM-95 Curcumin in context of DNA mismatch repair (MMR) status and CSC activity in 3D cultures of CRC cells. Pre- treatment with BCM-95 curcumin significantly enhanced the effect of 5-FU on HCT116R and HCR116+ch3R cells, in contrast to 5-FU alone as evidenced by increased disintegration of colonospheres, enhanced apoptosis and by inhibiting their growth. Curcumin and/or 5-FU strongly affected MMR-deficient CRC cells in high density cultures; however, MMR-proficient CRC cells were more sensitive. These effects of curcumin in enhancing chemosensitivity to 5-FU were further supported by its ability to effectively suppress CSC pools as evidenced by decreased number of CSC marker positive cells. The results illustrate novel and previously unrecognized effects of curcumin in enhancing chemosensitization to 5-FU-based chemotherapy on DNA MMR- deficient and their chemo-resistant counterparts by targeting the CSC sub-population. [Shakibaei M, Buhrmann C, Kraehe P, Shayan P, Lueders C and Goel A. Curcumin chemosensitizes 5-Fluorouracil resistant MMR-deficient human colon cancer cells in high density cultures. PLoS ONE. 2014:9(1).]
69. Curcumin suppresses crosstalk between colon cancer stem cells and stromal fibroblasts in the tumor microenvironment: potential role of EMT.
    Objective: Interaction of stromal and tumor cells plays a dynamic role in initiating and enhancing carcinogenesis. In this study, we investigated the crosstalk between colorectal cancer (CRC) cells with stromal fibroblasts and the anti-cancer effects of curcumin and 5- Fluorouracil (5-FU), especially on cancer stem cell (CSC) survival in a 3D-co-culture model that mimics in vivo tumor microenvironment.
    Methods: Colon carcinoma cells HCT116 and MRC-5 fibroblasts were co-cultured in a monolayer or high density tumor microenvironment model in vitro with/without curcumin and/or 5-FU.
    Results: Monolayer tumor microenvironment co-cultures supported intensive crosstalk between cancer cells and fibroblasts and enhanced up-regulation of metastatic active adhesion molecules (b1-integrin, ICAM-1), transforming growth factor-b signaling molecules (TGF-b3, p-Smad2), proliferation associated proteins (cyclin D1, Ki-67) and epithelial- to-mesenchymal transition (EMT) factor (vimentin) in HCT116 compared with tumor mono-cultures. High density tumor microenvironment co-cultures synergistically increased tumor-promoting factors (NF-kB, MMP-13), TGF-b3, favored CSC survival (characterized by up-regulation of CD133, CD44, ALDH1) and EMT-factors (increased vimentin and Slug, decreased E-cadherin) in HCT116 compared with high density HCT116 mono-cultures. Interestingly, this synergistic crosstalk was even more pronounced in the presence of 5-FU, but dramatically decreased in the presence of curcumin, inducing biochemical changes to mesenchymal-epithelial transition (MET), thereby sensitizing CSCs to 5-FU treatment.
    Conclusion: Enrichment of CSCs, remarkable activation of tumor-promoting factors and EMT in high density co-culture highlights that the crosstalk in the tumor microenvironment plays an essential role in tumor development and progression, and this interaction appears to be mediated at least in part by TGF-b and EMT. Modulation of this synergistic crosstalk by curcumin might be a potential therapy for CRC and suppress metastasis. [Buhrmann C, Kraehe P, Lueders C, Shayan P, Goel A, Shakibaei M. Curcumin suppresses crosstalk between colon cancer stem cells and stromal fibroblasts in the tumor microenvironment: potential role of EMT. PLoS ONE. 2014;9(9): e107514.]
70. Comparative Bioavailability of Curcumin, Turmeric, and Biocurcumax™ in Traditional Vehicles using Non-Everted Rat Intestinal Sac Model. The bioavailability of curcumin from turmeric, Biocurcumax and as plain curcumin was investigated using conventional vehicles by a non-everted rat intestinal model. Results of ex vivo intestinal permeability studies showed an enhancement in the permeability of curcumin with increase in lipophilicity of the vehicle used. Maximum permeability of curcumin was obtained from corn oil (13.4%) followed by clarified butter (9.82%), milk (4.24%) and aqueous suspension (1.66%) in 8 h. Another very interesting and important observation was that the permeation of curcumin was more from turmeric and Biocurcumax than from plain curcumin. These studies strongly suggest that curcumin may be consumed as turmeric/Biocurcumax in lipophilic vehicles instead of plain curcumin for maximum beneficial effects. [Shishu MM. Comparative bioavailability of curcumin, turmeric, and Biocurcumax™ in traditional vehicles using non-everted rat intestinal sac model. J Funct Foods. 2010;2(1):60-65.]
    

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    Ex Vivo or Cellular Studies Using Curcumin Combinations [10]

71. Effects of anti-inflammatory and adaptogenic herbal extracts on gene expression of eicosanoids signaling pathways in isolated brain cells.
    Introductions: The adaptogens modulate expression of genes playing key roles in development of aging- related disorders, which are considered as low-grade systemic inflammatory conditions characterized by an imbalance between pro-and anti-inflammatory eicosanoids.
    Aim of the Study: We compared the effects of anti-inflammatory and adaptogenic plant extracts on the expression of genes involved in biosynthesis of eicosanoids with the purpose to find those plants, which selectively upregulated the expression of anti- inflammatory lipoxins signaling pathways and inhibited pro-inflammatory signaling pathways associated with biosynthesis of leukotrienes, prostaglandins and thromboxanes.
    Materials and Methods: We conducted transcriptome-wide RNA sequencing to profile gene expression alterations in T98G neuroglia cells upon treatment of plant extract and analyzed the relevance of deregulated genes to eicosanoids signaling pathways using in silico models. RESULTS: For the first time, we demonstrated that Rhodiola rosea, Withania somnifera and Eleutherococcus senticosus downregulate the expression of key genes (ALOX5AP, DPEP2, LTC4S) involved biosynthesis of leukotrienes A, B, C, D and E, resulting in inhibition of leukotriene signaling pathway suggesting their potential benefits in Alzheimer disease. The common feature for all tested anti-inflammatory plants extracts was related to downregulation of ALOX12, which was also associated with neuroprotective action of these medicinal plants as well as their potential benefits in neurodegenerative diseases. None of tested anti- inflammatory and adaptogenic plants selectively activated the ALOX15-mediated signaling pathway, which is associated with generation anti-inflammatory lipoxins. Almost all tested plants upregulated the expression of the prostaglandin E receptor 3 gene (PTGER3) suggesting their potential benefits in the treatment of cancer.
    Consclusion: Every single plant tested in this study revealed a specific “signature” on eicosanoid signaling-related gene expression, regardless of their common features as anti- inflammatory or adaptogenic activity. Further studies of the combination of Rhodiola with Withania (Adaptra) for the treatment of Alzheimer disease are required. [Panossian A, Seo EJ, Efferth T. Effects of anti-inflammatory and adaptogenic herbal extracts on gene expression of eicosanoids signaling pathways in isolated brain cells. Phytomedicine. 2019 Mar 10: 152881]
    Note: Curcugreen reviewed in this study-see full study for details.
72. Curcumin downregulates expression of opioid-related nociception receptor gene (OPRL1) in isolated neuroglia cells.
    Background: Curcumin (CC) exerts polyvalent pharmacological actions and multi-target effects, including pain relief and anti- nociceptive activity. In combination with Boswellia serrata extract (BS), curcumin shows greater efficacy in knee osteoarthritis management, presumably due to synergistic interaction of the ingredients.
    Aim: To elucidate the molecular mechanisms underlying the analgesic activity of curcumin and its synergistic interaction with BS.
    Methods: We performed gene expression profiling by transcriptome-wide mRNA sequencing in human T98G neuroglia cells treated with CC (Curamed®), BS, and the combination of CC and BS (CC-BS; Curamin®), followed by interactive pathways analysis of the regulated genes. Results: Treatment with CC and with CC-BS selectively downregulated opioid- related nociceptin receptor 1 gene (OPRL1) expression by 5.9-fold and 7.2-fold, respectively. No changes were detected in the other canonical opioid receptor genes: OPRK1, OPRD1, and OPRM1. Nociceptin reportedly increases the sensation of pain in supra-spinal pain transduction pathways. Thus, CC and CC-BS may downregulate OPRL1, consequently inhibiting production of the nociception receptor NOP, leading to pain relief. In neuroglia cells, CC and CC-BS inhibited signaling pathways related to opioids, neuropathic pain, neuroinflammation, osteoarthritis, and rheumatoid diseases. CC and CC-BS also downregulated ADAM metallopeptidase gene ADAMTS5 expression by 11.2-fold and 13.5-fold, respectively. ADAMTS5 encodes a peptidase that plays a crucial role in osteoarthritis development via inhibition of a corresponding signaling pathway. Conclusion: Here, we report for the first time that CC and CC-BS act as nociceptin receptor antagonists, selectively downregulating opioid- related nociceptin receptor 1 gene (OPRL1) expression, which is associated with pain relief. BS alone did not affect OPRL1 expression, but rather appears to potentiate the effects of CC via multiple mechanisms, including synergistic interactions of molecular networks. [Seo EJ, Efferth T, Panossian A. Curcumin downregulates expression of opioid-related nociception receptor gene (OPRL1) in isolated neuroglia cells. Phytomedicine. 20 September 2018; https://doi.org/10.1016/j.phymed.2018.090.202]
73. Novel molecular mechanisms for the adaptogenic effects of herbal extracts on isolated brain cells using systems biology.
    Introduction: Adaptogens are natural compounds or plant extracts that increase adaptability and survival of organisms under stress. Adaptogens stimulate cellular and organismal defense systems by activating intracellular and extracellular signaling pathways and expression of stress-activated proteins and neuropeptides. The effects adaptogens on mediators of adaptive stress response and longevity signaling pathways have been reported, but their stress-protective mechanisms are still not fully understood.
    Aim of the Study: The aim of this study was to identify key molecular mechanisms of adaptogenic plants traditionally used to treat stress and aging-related disorders, i.e., Rhodiola rosea, Eleutherococcus senticosus, Withania somnifera, Rhaponticum carthamoides, and Bryonia alba.
    Materials and Methods: To investigate the underlying molecular mechanisms of adaptogens, we conducted RNA sequencing to profile gene expression alterations in T98G neuroglia cells upon treatment of adaptogens and analyzed the relevance of deregulated genes to adaptive stress-response signaling pathways using in-silico pathway analysis software.
    Results and Discussion: At least 88 of the 3516 genes regulated by adaptogens were closely associated with adaptive stress response and adaptive stress-response signaling pathways (ASRSPs), including neuronal signaling related to corticotropin-releasing hormone, cAMP-mediated, protein kinase A, and CREB; pathways related to signaling involving CXCR4, melatonin, nitric oxide synthase, GP6, Gαs, MAPK, neuroinflammation, neuropathic pain, opioids, renin–angiotensin, AMPK, calcium, and synapses; and pathways associated with dendritic cell maturation and G-coupled protein receptor–mediated nutrient sensing in enteroendocrine cells. All samples tested showed significant effects on the expression of genes encoding neurohormones CRH, GNRH, UCN, G- protein–coupled and other transmembrane receptors TLR9, PRLR, CHRNE, GP1BA, PLXNA4, a ligand-dependent nuclear receptor RORA, transmembrane channels, transcription regulators FOS, FOXO6, SCX, STAT5A, ZFPM2, ZNF396, ZNF467, protein kinases MAPK10, MAPK13, MERTK, FLT1, PRKCH, ROS1, TTN), phosphatases PTPRD, PTPRR, peptidases, metabolic enzymes, a chaperone (HSPA6), and other proteins, all of which modulate numerous life processes, playing key roles in several canonical pathways involved in defense response and regulation of homeostasis in organisms. It is for the first time we report that the molecular mechanism of actions of melatonin and plant adaptogens are alike, all adaptogens tested activated the melatonin signaling pathway by acting through two G-protein–coupled membrane receptors MT1 and MT2 and upregulation of the ligand-specific nuclear receptor RORA, which plays a role in intellectual disability, neurological disorders, retinopathy, hypertension, dyslipidemia, and cancer, which are common in aging. Furthermore, melatonin activated adaptive signaling pathways and upregulated expression of UCN, GNRH1, TLR9, GP1BA, PLXNA4, CHRM4, GPR19, VIPR2, RORA, STAT5A, ZFPM2, ZNF396,
FLT1, MAPK10, MERTK, PRKCH, and TTN, which were commonly regulated by all adaptogens tested. We conclude that melatonin is an adaptation hormone playing an important role in regulation of homeostasis. Adaptogens presumably worked as eustressors (“stress-vaccines”) to activate the cellular adaptive system by inducing the expression of ASRSPs, which then reciprocally protected cells from damage caused by distress. Functional investigation by interactive pathways analysis demonstrated that adaptogens activated ASRSPs associated with stress-induced and aging-related disorders such as chronic inflammation, cardiovascular health, neurodegenerative cognitive impairment, metabolic disorders, and cancer.
    Conclusions: This study has elucidated the genome-wide effects of several adaptogenic herbal extracts in brain cells culture. These data highlight the consistent activation of ASRSPs by adaptogens in T98G neuroglia cells. The extracts affected many genes playing key roles in modulation of adaptive homeostasis, indicating their ability to modify gene expression to prevent stress-induced and aging-related disorders. This study provides a comprehensive look at molecular mechanisms by which adaptogens exerts stress-protective effects. [Panossian A, Seo EJ, Efferth T. Novel molecular mechanisms for the adaptogenic effects of herbal extracts on isolated brain cells using system biology. Phytomedicine. 17Sep 2018;50:257-284.]
    Note: Curcugreen reviewed in this study-see full study for details.
74. A combination of curcumin and oligomeric proanthocyanidins offer superior anti- tumorigenic properties in colorectal cancer. Combining anti-cancer agents in cancer therapies is becoming increasingly popular due to improved efficacy, reduced toxicity and decreased emergence of resistance. Here, we test the hypothesis that dietary agents such as oligomeric proanthocyanidins (OPCs) and curcumin cooperatively modulate cancer-associated cellular mechanisms to inhibit carcinogenesis. By a series of in vitro assays in colorectal cancer cell lines, we showed that the anti-tumorigenic properties of the OPCs-curcumin combination were superior to the effects of individual compounds. By RNA-sequencing based gene-expression profiling in six colorectal cancer cell lines, we identified the cooperative modulation of key cancer-associated pathways such as DNA replication and cell cycle pathways. Moreover, several pathways, including protein export, glutathione metabolism and porphyrin metabolism were more effectively modulated by the combination of OPCs and curcumin. We validated genes belonging to these pathways, such as HSPA5, SEC61B, G6PD, HMOX1 and PDE3B to be cooperatively modulated by the OPCs-curcumin combination. We further confirmed that the OPCs-curcumin combination more potently suppresses colorectal carcinogenesis and modulated expression of genes identified by RNA-sequencing in mice xenografts and in colorectal cancer patient-derived organoids. Overall, by delineating the cooperative mechanisms of action of OPCs and curcumin, we make a case for the clinical co- administration of curcumin and OPCs as a treatment therapy for patients with colorectal cancer. [Ravindranathan P, et al. A combination of curcumin and oligomeric proanthocyanidins offer superior anti-tumorigenic properties in colorectal cancer. Sci Rep. 2018 Sep 14;8(1):13869.]
75. BCM-95 and (2-hydroxypropyl)-β-cyclodextrin reverse autophagy dysfunction and deplete store lipids in Sap C-deficient fibroblasts. Saposin (Sap) C deficiency is a rare variant form of Gaucher disease (GD) caused by impaired Sap C expression or accelerated degradation, and associated with accumulation of glucosylceramide (GC) and other lipids in the endo/lysosomal compartment. No effective therapies are currently available for the treatment of Sap C deficiency. We previously reported that a reduced amount and enzymatic activity of cathepsin (Cath) B and Cath D, and defective autophagy occur in Sap C-deficient fibroblasts. Here, we explored the use of two compounds, BCM-95, a curcumin derivative, and (2-hydroxypropyl)-β-cyclodextrin (HP- β-CD), to improve lysosomal function of Sap C-deficient fibroblasts. Immunofluorescence and biochemical studies documented that each compound promotes an increase of the expression levels and activities of Cath B and Cath D, and efficient clearance of cholesterol (Chol) and ceramide (Cer) in lysosomes. We provide evidence that BCM-95 and HP-β-CD enhance lysosomal function promoting autophagic clearance capacity and lysosome reformation. Our findings suggest a novel pharmacological approach to Sap C deficiency directed to treat major secondary pathological aspects in this disorder. [Tatti M, Motta M, Scarpa S, Di Bartolomeo S, Cianfanelli V, Tartaglia M, Salvioli R. BCM-95 and (2-hydroxypropyl)-β-cyclodextrin reverse autophagy dysfunction and deplete store lipids in Sap C-deficient fibroblasts. Hum Mol Genet. 2015 Aug 1;24(15):4198-4211.]
76. Synergistic and Additive Effects of Modified Novel evidence for curcumin and boswellic acid induced chemoprevention through regulation of miR-34a and miR- 27a in colorectal cancer. Colorectal cancer (CRC) is one of the most common causes of cancer-associated mortality worldwide, but it is truly a preventable disease. Both curcumin and boswellic acids are well-established dietary botanicals with potent anti-tumorigenic properties which have been shown to modulate multiple oncogenic pathways. Recent data suggest that the chemopreventive effects of these botanicals may in part be mediated through regulation of key cancer-related microRNAs (miRNAs) and their downstream gene targets. Here, we investigated the anti-tumorigenic effects of curcumin and 3 acetyl-11-keto-β-boswellic acid (AKBA) on modulation of specific cancer-related miRNAs in CRC cells and validated their protective effects in vivo using a xenograft mouse model. Both curcumin and AKBA inhibited cellular proliferation, induced apoptosis and cell cycle arrest in CRC cell lines, and these effects were significantly enhanced with combined treatment. Gene-expression arrays revealed that curcumin and AKBA regulated distinct cancer signaling pathways including key cell-cycle regulatory genes. Combined bioinformatics and in-silico analysis identified apoptosis, proliferation and cell-cycle regulatory signaling pathways as key modulators of curcumin and AKBA-induced anti-cancer effects. We discovered that curcumin and AKBA induced upregulation of tumor-suppressive miR-34a and downregulation of miR- 27a in CRC cells. Furthermore, we demonstrated in a mouse xenograft model that both curcumin and AKBA treatments suppressed tumor growth, which corresponded with alterations in the expression of miR-34a and miR-27a, consistent with our in vitro findings. Herein we provide novel mechanistic evidence for the chemopreventive effects of curcumin and AKBA through regulation of specific miRNAs in colorectal cancer. [Toden S, Okugawa Y, Buhrmann C, Nattamai D, Anguiano E, Baldwin N, Shakibaei M, Boland CR, Goel A. Cancer Prev Res (Phila). 2015 Feb 23. (Epub ahead of print).]
77. Citrus Pectin With Two Polybotanical Compounds, in the Suppression of Invasive Behavior of Human Breast and Prostate Cancer Cells.
    Aim: The objective of this study was to evaluate the combined effect of a known galectin-3 inhibitor, PectaSol-C modified citrus pectin (MCP), and 2 novel integrative polybotanical compounds for breast and prostate health, BreastDefend (BD) and ProstaCaid (PC), on invasive behavior in human breast and prostate cancer cells in vitro, respectively.
    Methods: The effect of MCP and BD and of MCP and PC on invasiveness was assessed by cell adhesion, cell migration, and cell invasion assays. Secretion of urokinase plasminogen activator (uPA) was determined by Western blot analysis.
    Results: Although low concentrations of MCP (0.25-1.0 mg/mL) do not suppress cell adhesion of breast or prostate cancer cells, the combination of MCP with BD or PC synergistically inhibits adhesion of these cells. Dose-dependent inhibition of breast and prostate cancer cell migration by MCP (0.25-1.0 mg/mL) is synergistically enhanced by BD (20 μg/mL) and PC (10 μg/mL), respectively. BD or PC did not further inhibit the invasion of breast and prostate cancer cells by MCP; however, the combination of MCP with BD or PC suppressed secretion of uPA from breast and prostate cancer cells, respectively. Conclusion. The combination of MCP with BD and of MCP with PC synergistically inhibits the metastatic phenotypes of human breast and prostate cancer cells, respectively. Further studies confirming these observations in animal models of breast and prostate cancer metastasis are warranted. [Jiang J, Eliaz I, Sliva D. Synergistic and Additive Effects of Modified Citrus Pectin With Two Polybotanical Compounds, in the Suppression of Invasive Behavior of Human Breast and Prostate Cancer Cells. Integrative Cancer Therapies. 2012;12(2):145-152.]
78. Suppression of Proliferation and Invasive Behavior of Human Metastatic Breast Cancer Cells by Dietary Supplement BreastDefend.
    Aim: The study was to evaluate the effect of the dietary supplement BreastDefend (BD) on the proliferation and invasive behavior of highly metastatic human breast cancer cells in vitro.
    Methods: Cell proliferation and cytotoxicity of BD was evaluated in MDA-MB-231 cells treated with BD (0-40 μg/mL) by MTT assay and trypan blue staining, respectively. Expression of cell cycle regulatory genes were determined by DNA-microarray analysis. Effect of BD on invasiveness was assessed by cellular adhesion, migration, and invasion assays.
    Results: BD treatment of cells MDA-MB-231 resulted in the cytostatic inhibition of cell proliferation with IC50 22.2, 19.1, and 17.5 μg/mL for 24, 48, and 72 hours, respectively. The inhibition of proliferation was mediated by the upregulation expression of CCNG1, CHEK1, CDKN1C, GADD45A, and E2F2, whereas BD downregulated expression of CCNA1 and CDK6 genes. The induction of expression of GADD45A and inhibition of expression of cyclin A1 (gene CCNA1) by BD was also confirmed on the protein level. BD treatment suppressed the invasive behavior of MDA-MB-231 cells by the inhibition of cellular adhesion, migration, and invasion. This inhibition of invasiveness was mediated by the suppression of secretion of urokinase plasminogen activator (uPA), and by the downregulation of expression of CXCR4 in breast cancer cells treated with BD.
    Conclusion: BD inhibits proliferation and invasive behavior of the highly metastatic human breast cancer cells in vitro. BD may have a therapeutic potential for prevention or treatment of highly metastatic breast cancers. [Jiang J, Wojnowski R, Jedinak A, Sliva D. Suppression of Proliferation and Invasive Behavior of Human Metastatic Breast Cancer Cells by Dietary Supplement BreastDefend. Integrative Cancer Therapies. Jun 2011;10(2):192-200.]
79. Suppression of growth and invasive behavior of human prostate cancers cells by ProstaCaid™: Mechanism of activity. Since the use of dietary supplements as alternative treatments or adjuvant therapies in cancer treatment is growing, a scientific verification of their biological activity and the detailed mechanisms of their action are necessary for the acceptance of dietary supplements in conventional cancer treatments. In the present study we have evaluated the anti-cancer effects of dietary supplement ProstaCaid™ (PC) which contains mycelium from medicinal mushrooms (Ganoderma lucidum, Coriolus versi-color, Phellinus linteus), saw palmetto berry, pomegranate, pumpkin seed, green tea [40% epigallocatechin-3-gallate (EGCG)], Japanese knotweed (50% resveratrol), extracts of turmeric root (BCM-95®), grape skin, pygeum bark, sarsaparilla root, Scutellaria barbata, eleuthero root, Job’s tears, astragalus root, skullcap, dandelion, coptis root, broccoli, and stinging nettle, with purified vitamin C, vitamin D3, selenium, quercetin, citrus bioflavonoid complex, β sitosterolzinc, lycopene, α lipoic acid, boron, berberine and 3.3′-diinodolymethane (DIM). We show that PC treatment resulted in the inhibition of cell proliferation of the highly invasive human hormone refractory (independent) PC-3 prostate cancer cells in a dose- and time-dependent manner with IC50 56.0, 45.6 and 39.0 μg/ml for 24, 48 and 72 h, respectively. DNA-microarray analysis demonstrated that PC inhibits proliferation through the modulation of expression of CCND1, CDK4, CDKN1A, E2F1, MAPK6 and PCNA genes. In addition, PC also suppresses metastatic behavior of PC-3 by the inhibition of cell adhesion, cell migration and cell invasion, which was associated with the down-regulation of expression of CAV1, IGF2, NR2F1, and PLAU genes and suppressed secretion of the urokinase plasminogen activator (uPA) from PC-3 cells. In conclusion, the dietary supplement PC is a promising natural complex with the potency to inhibit invasive human prostate cancer. [Jiang J, Eliaz I, Sliva D. Suppression of growth and invasive behavior of human prostate cancer cells by ProstaCaid™: Mechanism of activity. Int J Oncology. Jun 2011;38(6):1675- 1682.]
80. ProstaCaid induces G2/M cell cycle arrest and apoptosis in human and mouse androgen-dependent and-independent prostate cancer cells. The anticancer effects of ProstaCaid, a novel integrative blend of vitamins, minerals, multiherb extracts, and derivatives, were tested in human and mouse androgen-dependent (AD) and -independent (AI) prostate cancer cell lines. ProstaCaid shows growth inhibitory effects on both human and mouse AD prostate cancer cells (LNCaP and CASP 2.1) and AI prostate cancer cells (PC3 and CASP 1.1) in a dose-/time-dependent manner. Consistently, long-term treatment with ProstaCaid also reduced colony formation capacities of prostate cancer cells. Flow cytometry assays revealed that ProstaCaid induces G2/M arrest and apoptosis in LNCaP and PC3 cells after 72 hours of treatment. Immunoblotting assay demonstrated that 25 microg/mL of ProstaCaid treatment resulted in (1) the reduction of cyclin D1, cyclin B1, and Cdc2 expression in a time-dependent way; (2) increase in p21(WAF1/Cip1) as early as 12 hours after the treatments in PC3 cells and reduction to base line at the 72-hour time point; and (3) repression of Bcl-2, BclxL, and induction of Bim as well as the cleavages of caspase-3 and poly(ADP-ribose) polymerase (PARP) at 72 hours of treatment, suggesting caspase-3-dependent apoptosis. Moreover, ProstaCaid suppressed activation of AKT and MAPK signaling pathways in PC3 and LNCaP cells by reducing phosphorylation levels of AKT, its downstream target S6 ribosomal protein and GSK3beta, and ERK1/2, respectively. In summary, these findings strongly suggest that ProstaCaid may be a potential chemopreventive and therapeutic agent for both AD and, more importantly, AI prostate cancer. [Yan J, Katz AE. ProstaCaid induces G2/M cell cycle arrest and apoptosis in human and mouse androgen-dependent and and- independent prostate cancer cells. Integr Cancer Ther. 2010 Jun;9(2):186-196.]
    

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    Additional Supporting Studies

81. Efficacy of Curcumin and Boswellia for knee osteoarthritis: Systematic review and meta-analysis.
    Purpose: The unfavorable safety profiles of commonly prescribed knee osteoarthritis (OA) treatments have led clinicians and patients to seek safer alternatives. Research has suggested that curcuminoid and boswellia formulations could moderate key inflammatory pathways that are associated with worsening symptoms and disease progression. We conducted a systematic review and meta-analysis to assess the efficacy and safety of these treatments vs. placebo or NSAIDs for knee OA.
    Methods: We searched Medline, EMBASE, Google Scholar, Web of Science and the Cochrane database from inception to February 21, 2018. We also hand searched reference lists and reviewed conference proceedings. We included randomized clinical trials (RCTs) comparing curcuminoid or boswellia formulations with placebo or NSAIDs for knee OA. We calculated standardized mean differences (SMD) or risk ratios (RR) for all relevant outcomes. Meta-analyses were conducted using random effects models. Heterogeneity was assessed using the I2 statistic.
    RESULTS: Eleven RCTs (N = 1009) were eligible for analysis. Study quality was low overall, and most included RCTs were conducted on fewer than 100 participants. Both curcuminoid and boswellia formulations were statistically significantly more effective than placebo for pain relief and functional improvement. There were no significant differences between curcuminoids or boswellia and placebo in safety outcomes. Curcuminoids showed no statistically significant differences in efficacy outcomes compared to NSAIDs; patients receiving curcuminoids were significantly less likely to experience gastrointestinal adverse events. No RCTs compared boswellia against approved NSAIDs.
    CONCLUSIONS: The results of our study suggest that curcuminoid and boswellia formulations could be a valuable addition to the knee OA treatment regimens by relieving symptoms while reducing safety risks. The current body of evidence is not adequate in size or quality to make any meaningful clinical practice recommendations. Further research through large, high quality RCTs probably investigating the synergistic effect of these products with other OA treatments is warranted. [Bannuru RR, Osani MC, Al-Eid F, Wang C. Efficacy of Curcumin and Boswellia for knee osteoarthritis: Systematic review and meta-analysis. Semin Arthritis Rheum. 2018 Dec;48(3):416-429.]
82. The Role of Curcumin Administration in Patients with Major Depressive Disorder: Mini Meta-Analysis of Clinical Trials. Major depression is a common, recurrent, and chronic disease that negatively affects the quality of life and increases the risk of mortality. Several studies have demonstrated that curcumin, the yellow-pigmented substance of the turmeric, possesses antidepressant properties. The aim of this review is to meta-analytically assess the antidepressant effect of curcumin in patients with major depressive disorders. We extensively searched the literature until August 2015. The random-effect model was used to calculate the pooled standardized difference of means (SMD). Subgroup analyses were also performed to examine the effect of different study characteristics on the overall model. Six clinical trials met the inclusion criteria. Overall, curcumin administration showed a significantly higher reduction in depression symptoms [SMD = -0.34; 95% confidence interval (CI) = -0.56, -0.13; p = 0.002]. Subgroup analyses showed that curcumin had the highest effect when given to middle- aged patients (SMD = -0.36; 95% CI = -0.59; -0.13; p = 0.002), for longer duration of administration (SMD = -0.40; 95% CI = -0.64, -0.16; p = 0.001), and at higher doses (SMD = -0.36; 95% CI = -0.59, -0.13; p = 0.002). The administration of new formulation of curcumin (BCM-95) had non-significantly higher effect on depression as compared with the conventional curcumin-piperine formula. We conclude that there is supporting evidence that curcumin administration reduces depressive symptoms in patients with major depression. [Al-Karawi D, Al Mamoori DA, Tayyar Y. The Role of Curcumin Administration in Patients with Major Depressive Disorder: Mini Meta-Analysis of Clinical Trials. Phytother Res. 2015 Nov 27. doi: 10.1002/ptr.5524. (Epub ahead of print).]
83. Curcumin for neuropsychiatric disorders: a review of in vitro, animal and human studies. Turmeric has been used in traditional medicine for centuries to treat a range of ailments. Its primary active constituent curcumin, can influence an array of biological activities. Many of these, such as its anti-inflammatory, antioxidant, neuroprotective, and monoaminergic effects are dysregulated in several neuropsychiatric disorders. In this systematic review, in vitro, animal, and human studies investigating the potential of curcumin as a treatment for neuropsychiatric disorders such as major depressive disorder, post-traumatic stress disorder (PTSD), obsessive-compulsive disorder (OCD), bipolar disorder, psychotic disorders, and autism are reviewed, and directions for future research are proposed. It is concluded that curcumin is a promising, natural agent for many of these conditions, however, further research utilizing robust, clinical designs are essential. The problem associated with the poor oral bioavailability of standard curcumin also requires consideration. Currently the greatest support for the efficacy of curcumin is for the treatment of major depressive disorder. [Lopresti AL. Curcumin for neuropsychiatric disorders: a review of in vitro, animal and human studies. Journal of Psychopharmacology. 2017 Mar;31(3):287-302.]
84. The Role of Turmerones on Curcumin Transportation and P-Glycoprotein Activities in Intestinal Caco-2 Cells. The rhizome of Curcuma longa (turmeric) is often used in Asia as a spice and as a medicine. Its most well-studied component, curcumin, has been shown to exhibit poor bioavailability in animal studies and clinical trials. We hypothesized that the presence of lipophilic components (e.g., turmerones) in turmeric extract would affect the absorption of curcumin. The effects of turmerones on curcumin transport were evaluated in human intestinal epithelial Caco-2 cells. The roles of turmerones on P-glycoprotein (P-gp) activities and mRNA expression were also evaluated. Results showed that in the presence of α- and aromatic turmerones, the amount of curcumin transported into the Caco-2 cells in 2 hours was significantly increased. α – Turmerone and verapamil (a P-gp inhibitor) significantly inhibited the efflux of rhodamine-123 and digoxin (i.e., inhibited the activity of P-gp). It is interesting that aromatic turmerone significantly increased the rhodamine-123 efflux and Pgp (MDR1 gene) mRNA expression levels. The effects of a- and aromatic turmerones on curcumin transport as well as P-gp activities were shown here for the first time. The presence of turmerones did affect the absorption of curcumin in vitro. These findings suggest the potential use of turmeric extract (including curcumin and turmerones), rather than curcumin alone, for treating diseases. [Yue GGL, et al. The Role of Turmerones on Curcumin Transportation and P-Glycoprotein Activities in Intestinal Caco-2 Cells. Journal of Medicinal Food. 2012;15(3):242-252.]
85. Oat Fiber As a Carrier for Curcuminoids. The curcuminoid-carrying potential of oat fiber was examined as a potential route to overcome the low aqueous solubility of curcuminoids. Aqueous dispersions of oat fiber were mixed with curcuminoids solubilized in ethanol to obtain curcuminoids−oat fiber (1% w/w) dispersions in aqueous ethanol (2% v/v). Centrifugation of the curcuminoids−oat fiber dispersions resulted in a supernatant (95.3% w/w: 0.11% w/w protein, 0.17% w/w β-glucan) and precipitate (4.74% w/w: 0.18% w/w protein, 0.11% w/w β-glucan) with the curcuminoids being almost equally partitioned into both fractions. Curcuminoids solubility in the supernatant was markedly greater than that in aqueous ethanol and water. The curcuminoids were in the amorphous state in the precipitated fraction and were more stable to degradation than the curcuminoids in the supernatant. These studies show the potential of oat fiber as a carrier for curcuminoids into functional foods. [Sayanjali S, Sanguansri L, Buckow R, Gras S, Augustin MA. Oat Fiber As a Carrier for Curcuminoids. J Agric. Food Chem. 2014;62:12172-12177.]
86. The Role of Turmerones on Curcumin Transportation and P-Glycoprotein Activities in Intestinal Caco-2 Cells. The rhizome of Curcuma longa (turmeric) is often used in Asia as a spice and as a medicine. Its most well-studied component, curcumin, has been shown to exhibit poor bioavailability in animal studies and clinical trials. We hypothesized that the presence of lipophilic components (e.g., turmerones) in turmeric extract would affect the absorption of curcumin. The effects of turmerones on curcumin transport were evaluated in human intestinal epithelial Caco-2 cells. The roles of turmerones on P-glycoprotein (P-gp) activities and mRNA expression were also evaluated. Results showed that in the presence of α- and aromatic turmerones, the amount of curcumin transported into the Caco-2 cells in 2 hours was significantly increased. α -Turmerone and verapamil (a P-gp inhibitor) significantly inhibited the efflux of rhodamine-123 and digoxin (i.e., inhibited the activity of P-gp). It is interesting that aromatic turmerone significantly increased the rhodamine-123 efflux and Pgp (MDR1 gene) mRNA expression levels. The effects of a- and aromatic turmerones on curcumin transport as well as P-gp activities were shown here for the first time. The presence of turmerones did affect the absorption of curcumin in vitro. These findings suggest the potential use of turmeric extract (including curcumin and turmerones), rather than curcumin alone, for treating diseases. [Yue GGL, et al. The Role of Turmerones on Curcumin Transportation and P-Glycoprotein Activities in Intestinal Caco-2 Cells. Journal of Medicinal Food. 2012;15(3):242-252.]
87. Oat Fiber As a Carrier for Curcuminoids. The curcuminoid-carrying potential of oat fiber was examined as a potential route to overcome the low aqueous solubility of curcuminoids. Aqueous dispersions of oat fiber were mixed with curcuminoids solubilized in ethanol to obtain curcuminoids−oat fiber (1% w/w) dispersions in aqueous ethanol (2% v/v). Centrifugation of the curcuminoids−oat fiber dispersions resulted in a supernatant (95.3% w/w: 0.11% w/w protein, 0.17% w/w β-glucan) and precipitate (4.74% w/w: 0.18% w/w protein, 0.11% w/w β-glucan) with the curcuminoids being almost equally partitioned into both fractions. Curcuminoids solubility in the supernatant was markedly greater than that in aqueous ethanol and water. The curcuminoids were in the amorphous state in the precipitated fraction and were more stable to degradation than the curcuminoids in the supernatant. These studies show the potential of oat fiber as a carrier for curcuminoids into functional foods. [Sayanjali S, Sanguansri L, Buckow R, Gras S, Augustin MA. Oat Fiber As a Carrier for Curcuminoids. J Agric. Food Chem. 2014;62:12172-12177.] 

05/04/2016

1. 1. Double-blind placebo controlled trial of the anxiolytic effects of a standardized Echinacea extract
      Abstract: Earlier studies suggested that specific Echinacea preparations might decrease anxiety. To further  study the issue, we performed a double blind, placebo controlled trial with a standardized Echinacea angustifolia root extract. Participants were volunteers scoring above 45 points on the state or on the trait subscale of the State Trait Anxiety Inventory (STAI). They were treated with 40 mg Echinacea or with placebo tablets twice daily for 7 days followed by a 3 week‐long washout period. Participants were also administered the Beck Depression Inventory (BDI) and the Perceived Stress Scale (PSS). In the Echinacea group, state anxiety scores decreased by approximately 11 points by the end of the treatment period, whereas the decrease was around 3‐points in the placebo group (p< 0.01). The effect maintained over the washout period. The difference from placebo was significant from the 7th day of treatment throughout. Changes were less robust with trait anxiety scores, but the preparation performed better than placebo in patients with high baseline anxiety. Neither BDI nor PSS scores were affected by the treatments. Adverse effects were rare and mild, and all were observed in the placebo group. These findings suggest that particular Echinacea preparations have significant beneficial effects on anxiety in humans. [Haller J, Krecsak L, Zambori J. Double-blind placebo controlled trial of the anxiolytic effects of a standardized Echinacea extract. Phytotherapy Research 2019;1-9.]
   2. The Anxiolytic Potential and Psychotropic Side Effects of an Echinacea Preparation in Laboratory Animals and Healthy Volunteers. We investigate that toxicity, psychotropic side effects and anxiolytic potential of and Echinacea angustifolia extract that produced promising effects in laboratory tests performed earlier. Rats were studied in the elevated plus-maze, conditioned fear, open-field, object recognition and conditioned place preference test. Toxicity was studied in rats after intragastric administration. The preparation decreased anxiety in the elevated plus-maze and ameliorated contextual conditioned fear. No lethality or behavioral signs of discomfort were noticed in rats treated with 1000 and 3000mg/kg Echinacea angustifolia. The extract was without effect in tests of locomotion (open-field), memory (object recognition) and rewarding potential (conditioned place preference) within a wide dose range. A pharmacological formulation based on the same Echinacea angustifolia extract was tested in human subjects. One or two tablets per day were administered for 1 week to healthy volunteers scoring high on the State-Trait Anxiety Inventory (STAI). The tablets contain 20mg of the plant extract. Data were collected using a structured self-assessment diary technique. The high dose (2 tablets per day) decreased STAI scores within 3 days in human subjects, an effect that remained stable for the duration of the treatment (7 days) and for the 2 weeks that followed treatment. The lower dose (1 tablet per day) did not affect anxiety significantly. [Haller J, et al. The Anxiolytic Potential and Psychotropic Side Effects of an Echinacea Preparation in Laboratory Animals and Healthy Volunteers. Phytother Res. 2013 Jan;27(1):54-61.]
   3. An investigation into the anxiety-relieving and mood-enhancing effects of Echinacea
      angustifolia (EP107™): A randomised, double-blind, placebo-controlled study. Background: The acute anxiolytic effects of the echinacea angustifolia extract (EP107TM) have been demonstrated in two previous human trials. The goals of this study were to
      examine the anxiolytic and mood-enhancing effects of echinacea angustifolia over a longer duration with a larger sample size. Methods: In this 6-week, 3-arm, parallel-group, doubleblind, randomised controlled trial, 108 adults with mild-to-moderately severe anxiety were recruited and randomised to receive either a placebo, 20 mg, or 40 mg of echinacea angustifolia, twice daily. Outcome measures included the Clinically Useful Anxiety Outcome Scale (CUXOS), Positive and Negative Affect Scale (PANAS), Short Form-36 (SF-36), and Bergen Insomnia Scale (BIS). Results: Based on data collected from 104 participants, both doses of echinacea were associated with overall reductions in anxiety, although improvements were not different from the placebo. However, both doses of echinacea were associated with greater improvements in the PANAS positive and negative affect scores, and SF-36 emotional wellbeing score compared to the placebo. Limitations: The positive improvements associated with echinacea were only identified via the secondary outcome measures and, therefore, require validation in future trials. Conclusions: The echinacea angustifolia extract (EP107TM) administered for 6 weeks at a dose of 40 and 80 mg daily was not associated with greater improvements in anxiety in adults with mild-to-moderately severe anxiety compared to the placebo. However, there were greater improvements in positive and
      negative affect, and emotional wellbeing, suggesting antidepressant effects. Further studies using clearly-defined populations and validated outcome measures will be useful in future trials. [Lopresti AL, Smith SJ. An investigation into the anxiety-relieving and moodenhancing effects of Echinacea angustifolia (EP107™): A randomised, double-blind, placebo-controlled study. J Affect Disord. 2021;293:229-237.]
   4. Possible Role of Fat Tissue in the pharmacokinetics of Dodeca-2E, 4E, 8Z, 10E/Ztetraenoic Acid Isobutylamides after Oral Administration of Echniacea angustifolia Extract in Rats. Alkamides are one of the most important constituents of lipophilic extracts of Echinacea angustifolia roots. These compounds play an important role in the versatile pharmacological actions of this plant. The present study aimed to compare the concentrations of isomeric dodeca-2E,4E,8Z,10E/Z-tetraenoic acid isobutylamides (DTAI) in brain and periepididymal fat tissues and blood plasma of rats. Thirty, 60, 240 and 720 min after the oral administration of E. angustifolia root extract, tissue and plasma concentrations were determined by reversed-phase HPLC with ESI-MS/MS detection. The calculated terminal t of the mixture of DTAI was 8.28 h, which indicates a relatively slow elimination. In the 0.5–4 h period the brain/plasma and fat/plasma concentration ratios were continuously above 3 and 18, respectively, followed by equilibrium at 12 h. Our results indicate substantial accumulation of alkamides in lipid-rich tissues, which presumably contributes to a maintained pharmacological action. [Jedlinszki N, Redei D, Haller J, Freund T, Hohmann J, Zupko I. Possible Role of Fat Tissue in the Pharmacokinetics of Dodeca-2E, 4E, 8Z, 10E/Z-tetraenoic Acid Isobutylamides after Oral Administration of Echinacea angustifolia Extract in Rats. Natural Products Communications, Volume 9 Issue 6:737-888]
   5. The Effect of Echinacea Preparations in Three Laboratory Tests of Anxiety: Comparison with Chlordiazepoxide. Echinacea preparations are traditionally used to treat upper respiratory infections and inflammation. No psychotropic effects of Echinacea have been reported so far, although some recently reported active constituents are behaviorally active. Prompted by these finding, the anxiolytic potential of five different Echinacea preparations was evaluated. Three of these decreased anxiety but two of them had a very narrow effect dose range. Only one extract decreased anxiety within a wide dose-range (3-8mg/kg). Anxiolytic effects were consistently seen in three different tests of anxiety, the elevated plus-maze, social interaction and shock-induced social avoidance test. No locomotor suppressant effects were seen at nay dose. Noteworthy, the doses that showed anxiolytic effects in the present study were much lower than those used in the laboratory models of the traditional indications; Chlordiazepoxide robustly decreased anxiety-like behavior in all texts but suppressed locomotion at higher doses. Perceived and real risks of conventional medications increase the demand for alternative therapies, provided that these are safe and efficient. Earlier evidence shows that Echinacea preparations have an excellent safety profile, where our finding suggest for the first time that certain preparations have a considerable anxiolytic potential. Further research is required to identify facts that differentiate efficient and inefficient preparations. [Haller J, et al. The Effect of Echinacea Preparations in Three Laboratory Tests of Anxiety: Comparison with Chlordiazepoxide. Phytother Res. 2010 Nov;24(11):1605-13.]
   6. The Effects of an Echinacea Preparation on Synaptic Transmission and the Firing Properties of CA1 Pyramidal Cells in the Hippocampus. Traditionally, Echinacea preparations are used as anti-inflammatory agents and immune-enhancers. In addition to these effects, their anxiolytic potency has been recognized recently in laboratory tests. Our aim in this study was to uncover the potential effects of an Echinacea preparation on neuronal operations in the hippocampus, a brain region this is involved in anxiety and anxiety-related behaviors. Using in vitro electrophysiological techniques, we observed that excitatory synaptic transmission is hippocampal slices was significantly suppressed by an Echinacea extract found to be effective in anxiety tests. In contrast, no change in inhibitory synaptic transmission could be detected upon application on this extract. In addition, our experiments revealed that at low concentration the Echinacea extract reduced the spiking activity of CA1 pyramidal cells, while at high concentration increased it. This later observation was parallel to the reduction in the magnitude of the h-current-mediated voltage responses in pyramidal cells. At any concentrations, the passive membrane properties of CA1 pyramidal cells were found to be unaltered by the Echinacea extract. In summary, the Echinacea extract can significantly regulate excitatory, but not inhibitory, synaptic transmission in the hippocampus, and this action might be involved in its anxiolytic effects observed in behavior tests. [H jos N, et al. The Effects of an Echinacea Preparation on Synaptic Transmission and the Firing Properties of CA1 Pyramidal Cells in the Hippocampus. Phytother Res. 2012 Mar;26(3):354-62.]
   7. Alkamides and a neolignan from Echinacea purpurea roots and the interaction of alkamides with G-protein-coupled cannabinoid receptors. Multiple chromatographic separations of the CHCL3-soluble extract of the roots of Echinacea purpurea led to the isolation of 19 compounds. Four natural products, three alkamides and nitidanin diisovalerianate, were identified, and five further compounds were detected for the first time in the spices. Additionally, 10 known Echinacea purpurea metabolites were isolated. The structures were determined by mass spectrometry and advanced 1D and 2D NMR techniques. The bioactivity of the isolated compounds was studied in the [35S] GTPγS-binding experiments performed on rat brain membrane preparations. Both partial and inverse agonist compounds for cannabinoid (CB1) receptors were identified among the metabolites, characterized by weak to moderate interactions with the G-protein signaling mechanisms. The G-protein-modulating activities of the Echinacea compounds are rather far from the full agonist effects seen with the CB1 receptor agonist reference compound arachidonyl-2’-chloroethylamide (ACEA). However, upon coadministration with ACEA, a number of them proved capable of inhibiting the stimulation of the pure agonist, thereby demonstrating cannabinoid receptor antagonist properties. [Hohmann J, et al. Alkamides and a neolignan from Echinacea purpurea roots and the interaction of alkamides with G-protein-coupled cannabinoid receptors. Phytochemistry 2011 Oct;72(14-15):1848-53.]

1. Effects of N-acetylcysteine, oral glutathione (GSH) and a novel sublingual form of GSH on oxidative stress markers: A comparative crossover study. Glutathione (GSH) is critical to fight against oxidative stress. Its very low bioavailability limits the interest of a supplementation. The purpose of this study was to compare the bioavailability, the effect on oxidative stress markers and the safety of a new sublingual form of GSH with two commonly used dietary supplements, N-acetylcysteine (NAC) and oral GSH. The study was a three-week randomized crossover trial. 20 Volunteers with metabolic syndrome were enrolled. GSH levels and several oxidative stress markers were determined at different times during each 21-days period. Compared to oral GSH group, an increase of total and reduced GSH levels in plasma and a higher GSH/GSSG ratio (p=0.003) was observed in sublingual GSH group (supplementation with sublingual GSH demonstrated a more significant increase in the GSH/GSSG ratio than other forms of glutathione or precursors, +65% in relation to N-Acetylcysteine, +230% in relation to reference GSH). After 3 weeks of administration, there was a significant increase of vitamin E level in plasma only in sublingual GSH group (0.83 μmol/g; p=0.04). Our results demonstrate the superiority of a new sublingual form of GSH over the oral GSH form and NAC in terms of GSH supplementation. [Schmitt B, Vicenzi M, Garrel C, Denis FM. Effects of N-acetylcysteine, oral glutathione (GSH) and a novel sublingual form of GSH on oxidative stress markers: A comparative crossover study. Redox Biology. 2015;6:198-205.]

July 2016

Abstracts on omega-3s from salmon

1. 1. Anxiolytic-like effect of a salmon phospholipopeptidic complex composed of polyunsaturated fatty acids and bioactive peptides.
      A phospholipopeptidic complex obtained by the enzymatic hydrolysis of salmon heads in green conditions, exert anxiolytic-like effects in a time and dose-dependent manner, with no affection of locomotor activity. This study focused on the physic-chemical properties of the lipidic and peptidic fractions from this natural product. The characterization of mineral composition, amino acid and fatty acids was carried out. Stability of nanoemulsions allowed us to realize a behavioral study conducted with four different tests on 80 mice. This work highlighted the dose dependent effects of the natural complex and its various fractions over a period of 14 days compared to a conventional anxiolytic. The intracellular redox status of neural cells was evaluated in order to determine the free radicals scavenging potential of these products in the central nervous system (CNS), after mice sacrifice. The complex peptidic fraction showed a strong scavenging property and similar results were found for the complex as well as its lipidic fraction. For the first time, the results of this study showed the anxiolytic-like and neuroprotective properties of a phospholipopeptidic complex extracted from salmon head. The applications on anxiety disorders might be relevant, depending on the doses, the fraction used and the chronicity of the supplementation. [Belhaj N, Desor F, Gleizes C, et al. Anxiolytic-like effect of a salmon phospholipopeptidic complex composed of polyunsaturated fatty acids and bioactive peptides. Marine Drug. 2013;11:4294-4317.]
   2. Polar lipids: n-3 PUFA carriers for membranes and the brain: Nutritional interest and emerging processes.
      The n-3 fatty acids are unanimously considered to have high nutritional value, especially the long chain (LC) polyunsaturated fatty acids (PUFA) from marine origin. However, most of the products available in the market contain LC-PUFA extirified on the glycerol under the common form of triglycerols. The study investigates a process by which the PUFA can be esterified on polar lipids, especially phospholipids. The patented process is performed under low temperature without use of solvents and produces a Phospho-Lipo-Peptidic Complex that is particularly rich in DHA and is esterified in the proper sn-2 position on PL. Further, it has been demonstrated that n-3 fatty acids esterified on polar lipids instead of triglycerols, and retained in the proper, natural sn-2 position, have dramatically enhanced gastrointestinal absorption and intracellular incorporation (Bourre 2004). In pharmacokinetically validated CACO-2 studies, n-3 PUFA absorption across the enteric barrier was 5 fold over triglycerol bound n-3 PUFA fish oil. Subsequent intracellular levels were 50 fold greater. Phospholipids themselves have been shown in other studies (Kidd 1999; Bernoud, et al, 1999) to have direct brain impact, especially in the area of memory and learning performances in the aging human. Therefore, a process by which n-3 PUFA can be delivered in the correct sn-2 position and esterified to phospholipids may be a preferred delivery model over n-3 PUFA delivery esterified to triglycerols in fish oil. [Parmentier M, Mahmoud C, Linder M, Fanni J. Polar lipids: n-3 PUFA carriers for membranes and the brain: Nutritional interest and emerging processes. Oléagineux, Corps Gras, Lipides; Volume 14 (Issue 3): p.224-229.]

   ADDITIONAL INFORMATION
   Marine Phospho-Lipo-Peptidic complex for health benefits. In recent years, marine phospholipids represent a new source of n-3 fatty acids, in particular DHA for application in foods, neutraceuticals and cosmetics. According to the commercial terminology marine lecithin is used as a broad definition to describe a mixture of polar lipids (phospholipids and glycolipids) comprising predominantly long chain and highly unsaturated fatty acids obtained from a marine raw material. Phospholipids are major constituents of biological cell membranes. These amphiphilic molecules have a structural and functional synergy with fatty acids. Those are the lipophilic moiety associated with an hydrophilic part in form of a phosphoric acid ester. Phospholipids mainly differ by the group linked to the phosphoric ester (e.g. choline, ethanolamine, inositol, serine, …). The metabolic efficiency of these molecules depends on the type and location of the unsaturation of fatty acids. Marine phospholipids are characterized by very high levels of mostly long chain and highly unsaturated fatty acids mainly eicosapentaenoic and docosahexaenoic acids (C20:5 n-3 and C22:6 n-6, respectively). Docosahexaenoic acid (DHA), the end product of the omega-3 family fatty acid, is an abundant component in the brain phospholipids and a major nutrient of marine lipids. DHA is particularly enriched in the brain, retina, and spermatozoa phospholipids and plays a crucial role in brain development, learning ability, and visual acuity. [Linder M. Marine phosphor-lipo-peptidic complex for health benefits. White paper.]

   Analysis of lipids extracted from salmon (Salmo salar) heads by commercial proteolytic enzymes. Fresh salmon heads were submitted to controlled proteolysis using food-grade commercial enzymes (Alcalase, Neutrase, and Protamex). The release of oil under mild conditions (60°, 2h) compared favourably with organic solvent extraction (1.98% vs. 21.5%). Lipids extracted by solvent and lipids resulting from enzymatic processes displayed a similar content of PUFA (about 35%), mainly eicosapentaenoic acid (EPA; 8.4% vs. 7.7%) and docosahexaenoic acid (DHA; 12.1% vs 11.9%). Thin-layer chromatography (TLC-FID latroscan) showed that the polar lipid fraction accounted for 55% of total lipids (phophatidylethanolamine, 20.7%; phosphatidylcholine, 14.8%). Salmon head phospholipids may be more effective carriers of highly unsaturated fatty acids to specific tissues than triacyglycerols, as shown by their content in EPA (10.3 and 6.9%, respectively) and DHA (33.1 and 9.1%, respectively). [Gbogouri GA, Linder M, Fanni J, Parmentier M. Analysis of lipids extracted from salmon (Salmo salar) heads by commercial proteolytic enzymes. Eur J Lipid Sci Technol. 2006;108:766-775.]

   Doxosahexaenoic acid prevents neuronal apoptosis induced by soluble amyloid-β oligomers. A growing body of evidence supports the notion that soluble oligomers of amyloid-β (Aβ) peptide interact with the neuronal plasma membrane, leading to cell injury and inducing death-signaling pathways that could account for the increased neurodegeneration occurring in Alzheimer’s disease (AD). Doxosahexaenoic acid (DHA, C22:6, n-3) is an essential polyunsaturated fatty acid in the CNS and has been shown in several epidemiological and in vivo studies to have protective effects against AD and cognitive alterations. However, the molecular mechanisms involved remain unknown. We hypothesized that DHA enrichment of plasma membranes could protect neurons from apoptosis induced by soluble Aβ oligomers. DHA pre-treatment was observed to significantly increase neuronal survival upon Aβ treatment by preventing cytoskeleton perturbations, caspase activation and apoptosis, as well as by promoting extracellular signal-related kinase (ERK)-related survival pathways. These data suggest that DHA enrichment probably induced changes in neuronal membrane properties with functional outcomes, thereby increasing protection from soluble Aβ oligomers. Such neuroprotective effects could be of major interest in the prevention of AD and other neurodengenerative diseases. [ Florent S, Malaplate-Armond C, Youssef I, et al. Doxosahexaenoic acid prevents neuronal apoptosis induced by soluble amyloid-β oligomers. J Neurochem. 2006;96:385-395.]

02/16/15

Clinical studies of sea buckthorn

1. Effects of sea buckthorn oil intake on vaginal atrophy in postmenopausal women: A randomized, double-blind, placebo-controlled study.
   Background: Vaginal atrophy, the thinning and drying of vaginal mucosa, is associated with menopause. The standard estrogen treatment is not suitable for all women. Objective: To investigate the effects of oral sea buckthorn (SB) oil supplementation on vaginal atrophy. Method: A total of 116 postmenopausal women experiencing symptoms of vaginal dryness, itching or burning were randomized to this placebo-controlled, double-blind study. Ninety-eight participants completed the intervention of three months, during which they consumed 3 g of SB or placebo oil daily. At the beginning and end, factors of vaginal health were scored by a gynecologist, vaginal pH and moisture were measured and vaginal health index was calculated. Symptoms of atrophy and menopause were evaluated at study visits and by daily logbooks. Serum samples were collected for the analysis of circulating lipids, liver enzymes and C-reactive protein. Results: Compared to placebo, there was a significantly better rate of improvement in the integrity of vaginal epithelium in the SB group when both compliant and noncompliant participants were included (odds ratio (OR) = 3.1, 95% CI 1.11–8.95). A beneficial trend was observed when only the compliant participants were included (OR = 2.9; 95% CI 0.99–8.35). There was a tendency (P = 0.08) toward better improvement of vaginal health index from baseline to the end in the SB group [(0.8 (SD 2.8)] compared to placebo [−0.1(SD 2.0)].Conclusions: SB oil showed beneficial effects on vaginal health, indicating it is a potential alternative for mucosal integrity for those women not able to use estrogen treatment for vaginal atrophy. [Larmo PS, Yang B, Hyssala J, Kallio HP, Erkkola R. Maturitas. Effects of sea buckthorn oil intake on vaginal atrophy in postmenopausal women: A randomized, double-blind, placebo-controlled study. 2014; Article in Press.]
2. Different berries and berry fractions have various but slightly positive effects on the variables associated to metabolic diseases in overweight and obese women.
   Dietary habits have a major role in obesity, type 2 diabetes and atherosclerotic cardiovascular diseases. In this study, we compared the effects of sea buckthorn (SB) and its fractions, and bilberries (BBs) on associated variables of metabolic diseases on overweight and obese women. In total, 110 female volunteers were recruited, and they followed four different berry diets (BB, SB, SB phenolic extract (SBe) and SB oil (SBo)) in a randomized order for 33-35 days. Each intervention was followed by a wash-out period of 30-39 days. Blood samples were drawn and physical measurements were performed after each period. Eighty volunteers completed the study. There was statistically significant decrease in waist circumference after BB (Δ, -1.2 cm; P = 0.041) and SB (Δ, -1.1 cm; P = 0.008) periods and also a small decrease in weight after BB diet (Δ, -0.2 kg; P = 0.028). Vascular cell adhesion molecule decreased after BB (Δ, -49.8 ng/ml; P = 0.002) and SBo (Δ, -66.1 ng/ml; P = 0.001) periods, and in intercellular adhesion molecule (ICAM) after SBe diet (Δ, -6.1 ng/ml; P = 0.028). Based on the results, it can be stated that different berries and berry fractions have various but slightly positive effects on the associated variables of metabolic diseases. [Lehtonen HM, Suomela JP, Tahvonen R, Yang B, Venojärvi M, Viikari J, Kallio H. Different berries and berry fractions have various but slightly positive effects on the variables associated to metabolic diseases in overweight and obese women. Eur J Clin Nutr. 2011;65:394–401.]
3. Effects of oral sea buckthorn oil on tear film fatty acids in individuals with dry eye.
   Evaporative dry eye is associated with meibomian gland dysfunction and abnormalities of the tear film lipids. Dry eye is known to be affected positively by intake of linoleic and γ-linolenic acids and n-3 fatty acids. Oral sea buckthorn (Hippophaë rhamnoides) (SB) oil, which contains linoleic and α-linolenic acids and antioxidants, has shown beneficial effects on dry eye. The objective was to investigate whether supplementation with SB oil affects the composition of the tear film fatty acids in individuals reporting dry eye. One hundred participants were randomized to this parallel, double-blind, placebo-controlled study, which 86 of them completed. The participants daily consumed 2 g of SB or placebo oil for 3 months. Tear film samples were collected at the beginning, during, and at the end of the intervention and 1 to 2 months later. Tear film fatty acids were analyzed as methyl esters by gas chromatography.There were no group differences in the changes in fatty acid proportions during the intervention (branched-chain fatty acids: P = 0.49, saturated fatty acids: P = 0.59, monounsaturated fatty acids: P = 0.53, and polyunsaturated fatty acids: P = 0.16). The results indicate that the positive effects of SB oil on dry eye are not mediated through direct effects on the tear film fatty acids. Carotenoids and tocopherols in the oil or eicosanoids produced from the fatty acids of the oil may have a positive effect on inflammation and differentiation of the meibomian gland cells. [Järvinen R, Larmo P, Setälä N, Yang B, Engblom J, Viitanen M, Kallio H. Effects of oral sea buckthorn oil on tear film fatty acids in individuals with dry eye. Cornea 2011;30:1013–1019 (Samples collected in the dry eye study of Larmo et al (2010) J Nutr).]
4. Oral sea buckthorn oil attenuates tear film osmolarity and symptoms in individuals with dry eye.
   Dry eye is a common condition that can severely impair the quality of life. We aimed to find out whether oral sea buckthorn (SB) oil, containing (n-3) and (n-6) fatty acids and antioxidants, affects dry eye. In this double-blind, randomized, parallel trial, 20- to 75-y-old women and men experiencing dry eye symptoms consumed 2 g of SB or placebo oil daily for 3 mo from fall to winter. One hundred participants were recruited and 86 completed the study. Clinical dry eye tests and symptom follow-ups were performed. Tear film hyperosmolarity is a focal factor in dry eye. There was a general increase in the osmolarity from baseline to the end of the intervention. Compared with the placebo group, the increase was significantly less in the SB group when all participants were included [intention to treat (ITT), P = 0.04] and when only participants consuming the study products for at least 80% of the intervention days were included [per protocol (PP), P = 0.02]. The maximum intensities of redness and burning tended to be lower in the SB group. In the ITT participants, the group difference was significant for redness (P = 0.04) but not for burning (P = 0.05). In the PP participants, the group difference was significant for burning (P = 0.04) but not for redness (P = 0.11). In conclusion, SB oil attenuated the increase in tear film osmolarity during the cold season and positively affected the dry eye symptoms. [Larmo P, Järvinen R, Setälä N, Yang B, Viitanen M, Engblom J, Tahvonen R, Kallio H. Oral sea buckthorn oil attenuates tear film osmolarity and symptoms in individuals with dry eye. J Nutr. 2010;140:1462-1468.]
5. Berry meals and risk factors associated with metabolic syndrome.
   Nonalcoholic fatty liver disease is commonly associated with obesity, insulin resistance, dyslipidemia and type 2 diabetes, and can thus be regarded as the hepatic manifestation of metabolic syndrome. In this study we compared the effects of lifestyle intervention with and without industrial berry products, on risk factors associated with metabolic syndrome on slightly overweight women. Sixty-one female volunteers (average age 42.9 years) were recruited and randomized for a 20-week dietary intervention trial with two parallel treatment groups, one lifestyle intervention group with berry products equaling with an average daily dose of 163 g of northern berries (berry group, diet 1, N=31, of which 28 completed the study) and the other group with lifestyle intervention only (control group, diet 2, N=30, of which 22 completed the study).Increased berry consumption as part of the normal daily diet was the only lifestyle difference between the two intervention groups. The major effects achieved by diet 1 were changes in the levels of alanine aminotransferase (ALAT) and adiponectin (at P-values <0.001 and 0.002, respectively). A statistically significant difference between the two intervention groups was the higher decrease in the ALAT value in the berry group (P=0.003). The 23% decrease in the ALAT value, from 20.29 to 15.66 U/l in the berry group may be regarded as nutritionally significant by enhancing the liver function. This may contribute positively to the low-grade systemic inflammation in body and decrease the risk of cardiovascular diseases. [Lehtonen HM, Suomela JP, Tahvonen R, Vaarno J, Venojärvi M, Viikari J, Kallio H. Berry meals and risk factors associated with metabolic syndrome. Eur J Clin Nutr. 2010;64:614-621]
6. Effects of oral supplementation and topical application of supercritical CO2 extracted sea buckthorn oil on skin ageing of female subjects.
   Sea buckthorn (Hippophaë rhamnoides) seed and pulp oil is known to promote skin regeneration, speed up wound healing and ease skin inflammation. The present study was carried out to investigate the anti-ageing effects of oral supplementation of a standardized supercritical CO2-extracted sea buckthorn oil and topical skin application of supercritical CO2-extracted sea buckthorn seed oil. Sixty female subjects of age 50-70 years were randomly divided into two groups, thirty subjects in each group. In one group, the subjects took sea buckthorn oil capsules, four capsules (4 x 0.5 g oil) per day, for three months. In the other group, the subjects applied topically sea buckthorn seed oil rejuvenating night cream on the face, twice per day, for three months. Skin hydration status, elasticity, surface roughness, luminosity and cutaneous thickness were determined using non-invasive instrumental measurements before, after one month and at the end of treatments. Both treatments significantly improved the skin hydration status and the overall skin elasticity of the subjects (P < 0.001). Oral supplementation with the sea buckthorn oil capsules resulted in decreases in the mean roughness (Ra) and maximum roughness (Rz) of the skin surface, indicating anti-wrinkle efficacy of the product. Topical application of the sea buckthorn oil cream increased cutaneous thickness, suggesting positive structural changes and improvement in collagen synthesis in the skin. This is the first study showing beneficial effects of orally used sea buckthorn oil in reducing the signs of skin ageing. This is also an important breakthrough demonstrating the great potential of sea buckthorn oil in supporting the health and well being of healthy skin through internal use. [Yang B, Bonfigli A, Pagani V, Isohanni T, von-Knorring Å, Jutila A, Judin V-P. Effects of oral supplementation and topical application of supercritical CO2 extracted sea buckthorn oil on skin ageing of female subjects. J. Appl. Cosmetol. 2009 Jan-Mar;27:1-13.]
7. Sea buckthorn oils, mucous membranes and Sjögren’s syndrome with special reference to latest studies.
   Source: Yang B, Erkkola R. (2008) In: Singh et al., editors. Seabuckthorn (Hippophae L.). A Multipurpose Wonder Plant. Vol. III: Advances in research and development. New Delhi, India: Dya Publishing House; p.254-267
8. Absorption of flavonols derived from sea buckthorn (Hippophaë rhamnoides L.) and their effect on emerging risk factors for cardiovascular disease in humans.
   Sea buckthorn (Hippophaë rhamnoides L.) is a rich source of flavonols, especially isorhamnetin. Most prospective cohort studies have indicated some degree of inverse association between flavonoid intake and coronary heart disease. Animal and human studies suggest that sea buckthorn flavonoids may scavenge free radicals, lower blood viscosity, and enhance cardiac function. The effects of flavonol aglycones derived from sea buckthorn on the risk factors of cardiovascular disease as well as their absorption were studied in humans. The flavonols, ingested with oatmeal porridge, did not have a significant effect on the levels of oxidized low-density lipoprotein, C-reactive protein, and homocysteine, on the plasma antioxidant potential, or on the paraoxonase activity. Flavonols at two dosages in oatmeal porridge were rapidly absorbed, and a relatively small amount of sea buckthorn oil added to the porridge seemed to have increased the bioavailability of sea buckthorn flavonols consumed at the higher dose. [Suomela JP, Ahotupa M, Yang B, Vasankari T, Kallio H. Absorption of flavonols derived from sea buckthorn (Hippophaë rhamnoides L.) and their effect on emerging risk factors for cardiovascular disease in humans. J Agric Food Chem. 2006;54:7364-7369.]
9. Sea Buckthorn oils: towards healthy mucous membranes.
   Topical application sea buckthorn oils and preparations containing sea buckthorn oil have been shown to improve the health of mucous membranes of urogenital tract. Therefore we decided to design a trial by using orally administered sea buckthorn oils for the treatment of patient who had suffered from conditions described above, and had experienced a multitude of treatments. The patients took orally capsules of Sea Buckthorn Oil (extracted by supercritical CO2, kindly provided by Aromtech Ltd, Finland), a mixture of oils from seeds and berry soft parts, 3 capsules twice a day (3 g sea buckthorn oil per day) for 12 weeks. The committee of ethics accepted the study plan. Significant improvement was seen in three severe cases after sea buckthorn oil treatment. The best improvement was seen in patient No.2 representing as much as 66% decrease of the total VAS score. The average total score value was decreased by 46%, from 185 to 100 by sea buckthorn oil treatment. Estrogen levels at the end of trial were equal to the pretreatment measurements. None of the subjects reported any side effects. [Erkkola R, Yang B. Sea Buckthorn oils: towards healthy mucous membranes. AgroFood Industry Hi-Tech. 2003;May-June.]
10. Sea buckthorn berry oil inhibits platelet aggregation.
    A small-scale preliminary cross-over study was conducted to investigate the effects of supercritical CO(2)-extracted sea buckthorn berry oil (SBO) on some risk factors of cardiovascular disease. Special features of the oil are high proportions of palmitic (16:0), oleic (18:1n-9), palmitoleic (16:1n-7), linoleic (18:2n-6), and alpha-linolenic (18:3n-3) acids as well as vitamin E, carotenoids, and sterols. Twelve healthy normolipidemic men were recruited and each volunteer consumed SBO and fractionated coconut oil (control) 5 g per day for a period of 4 weeks in a random order (wash-out 4-8 weeks). Phospholipid fatty acids, plasma lipids, and glucose were unaffected by SBO supplementation. Instead, a clear decrease in the rate of adenosine-5′-diphosphate-induced platelet aggregation and maximum aggregation were found. This suggested the beneficial effects of SBO on blood clotting, but further studies on the dose-response effects are needed to assess the practical use of SBO supplements. [Johansson AK, Korte H, Yang B, Stanley JC, Kallio HP. Sea buckthorn berry oil inhibits platelet aggregation. J Nutr Biochem. 2000;11:491-495.]