Prasad Arvind Thakurdesai

Objective: The efficacy and safety evaluation of oral “glycosides-based standardized fenugreek seed extract" (SFSE-G) supplementation to a healthy aging adult population. Methods: A total of 112 healthy individuals aged ≥ 45 years (determined by chronological age and Levine’s phenotypic age) were randomized using a computer-generated list (1:1 ratio) to receive 300 mg capsules of either SFSE-G or placebo daily for 12 weeks in a double-blind design. The outcome measures were changes in plasma “nicotinamide adenine dinucleotide” (NAD⁺) levels, plasma “adenosine triphosphate” (ATP) levels, phenotypic age, physical performance, fatigue, and quality of life, along with safety and tolerability. Results: Out of 112 participants enrolled, 103 completed the study (SFSE-G: 50; placebo: 53). Plasma NAD+ concentrations were significantly (P < 0.05) higher when age correlated in SFSE-G-treated groups (vs. placebo). No serious adverse events or safety concerns were reported, indicating good tolerability of treatments. In addition, the study found that 12 weeks of 300 mg SFSE-G oral supplementation significantly (P < 0.05) increased ATP levels, improved physical fatigue, and showed trends in reducing phenotypic age and improving quality of life in correlation with age. Conclusion: SFSE-G supplementation showed potential to mitigate age-related declines in energy metabolism and physical fatigue, with a robust safety profile.

Objective: This study investigated the effects of glycosides-rich standardized fenugreek seed extract (SFSE-G) in a mouse model of “unilateral ureteral obstruction” (UUO)-induced renal fibrosis. Methods: UUO was performed in 32 female mice (C57BL/6 strain) and randomized into groups of eight mice each. A separate group of eight mice (sham control) underwent sham operation with no UUO surgery. Mice were orally administered vehicle (distilled water) or SFSE-G at doses of 30, 60, or 100 mg/kg twice daily for 13 days. Various biochemical, histological, and gene expression-related measurements were conducted on day 14 after euthanasia, and body weights were measured daily. Results: Subacute oral administration of SFSE-G showed dose-dependent significance (p<0.05, P<0.01) to reduce UUO-induced elevation of blood urea nitrogen levels and attenuated histopathological changes, including tubular injury and collagen deposition. Quantitative PCR revealed that SFSE-G downregulated oxidative stress-and fibrosis-related gene expression of markers such as “α-smooth muscle action”, “nuclear factor erythroid 2-related factor 2”, and “heme oxygenase-1”. Although SFSE-G-treated mice showed a downward trend against UUO-induced increases in hydroxyproline content and gene expression of “transforming growth factor-β1” and tissue inhibitor of collagen type 1 and metalloproteinase-1, the differences were not statistically significant. Conclusion: SFSE-G exerts fibrosis preventive and renoprotective effects in UUO-induced renal fibrosis, probably by modulating oxidative stress and fibrotic pathways.

Objective: To evaluate developmental toxicity of the polyphenol-based standardized cinnamon bark extract (IND02) during the gestational period in pregnant rats using “Organization for Economic Co-operation and Development” (OECD) Test No. 414. Methods: Pregnant female rats were daily administered IND02 via gavage (125, 250, or 500 mg/kg) from Gestational Days (GD) 05 to GD19, except for the Vehicle Control (VC) group. On GD20, the dams underwent cesarean section, and observations were made of the uteri and fetuses of the dams. Results: Prenatal oral administration of IND02 during gestation period in female rats did not show detrimental effects on maternal (body weight, uterine weight, uterine morphology, and gross pathology) or fetal development parameters (crown-rump length, sex ratio, and occurrence of anomalies) compared with VC, indicating normal fetal development. A few skeletal and visceral malformations that were noted were infrequent, incidental, and without toxicological significance. Conclusion: Prenatal oral exposure of rats to IND02 did not result in developmental toxicity (fetotoxicity or teratogenicity). “No-Observed-Adverse-Effect Level” (NOAEL) and “Human Equivalent Dose” (HED) of IND02 in rats at prenatal oral exposure was more than 500 mg/kg/d and 4.86 g/d respectively.

Objective: To assess the effectiveness of nasal solution of Centella asiatica leaves standardized to triterpenoids (INDCA-NS) in preventing “chronic unpredictable mild stress” (CUMS) in rats. Methods: The study involved six groups of twelve rats each, with five groups receiving CUMS induction and one group without CUMS, serving as a vehicle control (VC). The other groups received intranasal administration of saline, buspirone, or INDCA-NS (10,30 and 100 µg/rat/day, 2.5, 7.5, and 10 µg/nostril/twice daily) from day 35 to day 49. Behavioral parameters were assessed using the marble burying test, Y-maze, Morris water maze, sucrose preference test, and resident intruder test on days 0, 35, 42, and 49, respectively. The levels of stress-related biomarkers, cortisol and “brain-derived neurotrophic factor” (BDNF) in the hypothalamus were measured using ELISA kits. The data was analyzed with analysis of variance followed by pairwise comparisons, with significance set at P < 0.05. Results: CUMS induction led to a significant increase in anxiety, anhedonia, aggression, and stress markers, as well as a reduction in working and spatial memory parameters (vs. VC). INDCA-NS and BUS administration for 14 days resulted in dose-dependent and significant prevention of CUMS-induced anxiety, working memory, anhedonia, and aggression, but not Morris’s water maze parameters and BDNF levels. Conclusion: Subacute intranasal INDCA-NS showed chronic stress-preventive potential in rats, indicated by the prevention of anxiety, anhedonia, and aggression in CUMS-induced rats, probably through cortisol reduction.

Objective: To investigate the prebiotic effects of defatted fenugreek seed flakes (Fenuflakes™) using a 48-hour colonic simulation model. Methods: Fecal samples from three healthy adult human donors were exposed to treatment conditions of blank (medium control), Fenuflakes, or inulin (fiber control). The effects on microbial fermentation (pH and gas production), metabolite production [Short Chain Fatty Acid (SCFAs), Branched-Chain Fatty Acid (BCFA), ammonium and lactate production], and microbial community composition were evaluated at 0, 6, 24, and 48 h. Results: In comparison to inulin, Fenuflakes demonstrated a significant reduction in gas production. Both inulin and Fenuflakes significantly increased beneficial SCFAs, but no significant change was observed in BCFA. Ammonium production increased upon Fenuflakes treatment due to a residual protein fraction reaching the colon, in contrast to inulin, which is in a protein-depleted state. The lactate levels significantly increased for Fenuflakes and inulin. The presence of Fenuflakes and inulin resulted in favorable fermentation by the colonic gut microbiota, indicating an increase in species enrichment and alpha diversity in the microbial community composition. Conclusion: Fenuflakes exhibit prebiotic potential by increasing SCFA and promoting the enrichment of several beneficial colonic bacteria while maintaining normal colonic pH and producing less gas than inulin.

The present study aimed to determine the efficacy of intranasal administration of a standardized extract of Gotu kola, i.e., Centella asiatica (L.) Urban (INDCA-NS) with marker triterpenoids for the prevention of nitroglycerine- (NTG)-induced recurrent migraine in rats. Adult rats of both sexes in a group of 12 were administered intraperitoneal NTG (10 mg/kg) on alternate days (D1 to D9) and once daily intranasal solutions of either vehicle (saline, 50 µL/rat/day), sumatriptan (80 µL/rat/day of 12 mg/ml) as positive control, or INDCA-NS (10, 30, or 100 µg/rat/day) for 21 days. Behavioral and biochemical parameters related to concurrent migraine pain (facial expressions on the grimace scale, thermal hyperalgesia, mechanical allodynia, and plasma and brain levels of pituitary adenylate cyclase-activating polypeptide and nitric oxide), and stress (photophobia and cortisol levels in the brain and serum) were measured. The intranasal administration of INDCA-NS prevented NTG-induced migraine-like pain, photophobia, and stress in a dose-dependent manner. At the same time, sumatriptan alleviated pain and anxiety but not photophobia. In conclusion, the intranasal administration of INDCA-NS showed prophylactic efficacy against recurrent NTG-induced migraine pain in rats.

Background: Cinnamon (Cinnamomum zeylanicum) bark is used as a spice in culinary practices and has been documented for health and medicinal benefits. Polyphenol (PP) is the major bioactive constituent of cinnamon bark. However, acceptable and safe dose levels and toxicity information of oral exposure to PP-based standardized cinnamon bark extract (PP-CZ) are crucial for safe human consumption. Objective: To evaluate PP-CZ for acute oral, subchronic oral toxicities in vivo, mutagenicity in vitro and genotoxic potential in vitro. Method:. The effects of oral treatment with single- and 90-days repeated dose were evaluated in rates as per OECD Test 423 and 408, respectively. Effects on body weight, food and water intake, organ weight, hematology, biochemistry, and histology were recorded . Mutagenicity and genotoxicity were evaluated using Ames (OECD No. 471) and chromosomal aberrations (OECD Test. 473) tests. Results: A single oral dose of PP-CZ did not cause death or treatment-related toxic effects, indicating a “median lethal dose” > 2,000 mg/kg. In addition, a subchronic dosage (500 mg/kg/day, 90 days) was found safe in rats, suggesting “no observed adverse effect level” (NOAEL) of 500 mg/kg and “Human Equivalent Dose” (HED) of 4.8 g/day. Furthermore, the absence of mutagenicity or genotoxicity of PP-CZ was observed during in vitro tests. PP-CZ showed a robust safety profile without mutagenicity or genotoxicity in rats.

The leaves of Centella asiatica L. Urban (C. asiatica) and their bioactive compounds, triterpenoids asiaticoside (AS) and madecassoside (MA), are effective in reducing psychological stress and associated behavioral disorders in the conducted in vivo research. The present study evaluated AS+MA-based standardized C. asiatica leaves extract (INDCA) on SIS-induced suicidal behavior-related traits in laboratory rats. Male rats (n=6) were randomized, grouped, and individually caged for seven days for stress induction. For the next seven days (D7 to D14), rats were orally administered vehicle (stress control), positive control (fluoxetine, 30 mg/kg), and or INDCA (3, 10, 30 mg/kg) once a day. A separate group of rats without isolation stress (normal rats) was maintained. The scores for suicidal behavior-related traits, such as aggression, impulsivity, irritability, learned helplessness, and plasma cortisol, were measured after 14-day treatment. The stress control group showed a significant increase in aggression, irritability (total score), learned helplessness (escape latency, escape failure, and recovery attempts), and plasma cortisol levels, which confirmed suicidal behavior-related traits. The INDCA-treated rats showed a dose-dependent reduction in stress-induced behavioral traits and elevated plasma cortisol levels. In conclusion, subacute administration of INDCA showed amelioration of suicidal behavior in social-isolation-induced stress in laboratory rats and suggested a promising natural and safe option for the management of stress-induced behavioral disorders, including suicidal behavior.