Toxicological safety assessment of AP-Bio®, a standardized extract of Andrographis paniculata in Sprague Dawley rats.

Andrographis paniculata, commonly known as green chiretta, is a traditionally used plant in India, China, and Southeast Asian countries for its varied health benefits including immune health. The objective of the present study was to assess the safety of AP-Bio®, a standardized A. paniculata extract in Sprague Dawley rats by following the Organisation for Economic Cooperation and Development (OECD) test guidelines of acute and 90-day repeated dose sub-chronic toxicity studies. AP-Bio® did not show any treatment-related clinical signs of toxicity or mortality during the 14-day observation period in animals tested in the single-dose acute oral toxicity study up to a dose of 5000 mg/kg body weight. In the 90-day repeated dose sub-chronic oral toxicity study, no treatment-related adverse clinical signs were observed in any of the treated groups (300, 600, and 900 mg/kg). All treated animals showed usual weight gain and comparable feed intake. The ophthalmoscope examination did not reveal any abnormalities. Also, no toxicologically significant changes were observed in urinalysis, hematology, and blood chemistry parameters. Absolute organ weights and relative organ weights of vital organs did not differ significantly compared to control. Gross and histopathological findings did not show any remarkable and treatment-related changes. Results of the safety evaluation showed the median lethal dose (LD50 ) of AP-Bio® was found to be more than 5000 mg/kg rat body weight and the no observed adverse effect level (NOAEL) of AP-Bio® was found to be 900 mg/kg rat body weight.

Ocimum tenuiflorum extract (HOLIXERTM): Possible effects on hypothalamic-pituitary-adrenal (HPA) axis in modulating stress.

Ocimum tenuiflorum is a sacred medicinal plant bestowed with multiple health benefits. This plant is traditionally considered an adaptogen. Many scientific studies have indicated the anti-stress potential of Ocimum tenuiflorum but with higher doses. The present study investigated the effects of HolixerTM (a clinically studied standardized Ocimum tenuiflorum extract) on modulating stress using two in vivo models, namely the swim endurance study in mice and forced swim test in rats. In addition, we explored the mechanism of action of HolixerTM on the HPA axis using two in vitro cell-based assays to check for its inhibitory effect on cortisol release and CRF1 receptor antagonistic activity. Ocimum tenuiflorum extract enhanced the swimming time in mice, reduced the stress-induced increase in immobility time, and prevented the increase in corticosterone in rats subjected to the forced swim test. Further, Ocimum tenuiflorum extract inhibited cortisol release and exhibited a significant CRF1 receptor antagonist activity. Thus, Ocimum tenuiflorum extract was found effective in managing stress, and the effect could be due to the inhibition of cortisol release and the antagonistic effect on the CRF1 receptors.

Identification of lipid regulatory genes modulated by polyherbal formulation in chicken liver tissues using transcriptome analysis.

Objective: To elucidate the cellular mechanisms of polyherbal formulation [Kolin PlusTM (KP)], genomics was performed to delineate the genes and pathways associated with lipid regulation through transcriptional profiling of the liver in commercial broilers raised on diets deficient in choline chloride (CCL). Materials and Methods: The gene expression patterns were studied for four groups [normal diet: normal, choline chloride deficient (CCD), KP (400 gm/ton), and CCL (400 gm/ton)] using Agilent microarray on day 42. The hierarchical cluster analysis was carried out on 12,614 differentially expressed genes (DEGs) with a similar expression. Results: Out of 12,614 significant DEGs, 1,926, 448, and 1,330 genes were expressed at higher rates, and 413, 482, and 1,364 were expressed at lower rates than CCD (CCD vs. normal), CCL (CCL vs. CCD), and KP (KP vs. CCD), respectively. GO enrichment analysis of DEG further revealed the significant association of biological process items with the lipid, sterol, and lipoprotein metabolic processes. In particular, peroxisome proliferator-activated receptor gamma coactivator 1 alpha, carnitine palmitoyl transferase I, hydroxyacyl-CoA dehydrogenase trifunctional multienzyme complex subunit beta, and patatin-like phospholipase domain containing 2 genes involved in fatty acid oxidation and lipase C, ABCG5, ABCG8, acetyl-CoA carboxylase, ATP citrate lyase enzyme, and peroxisome proliferator-activated receptor gamma genes involved in lipogenesis were altered by KP intervention for lipid metabolism. Conclusions: These findings reveal that the supplementation of KP prevents fatty liver-associated problems in broiler chickens by modulating the expression of the above-mentioned genes that are responsible for the oxidation of fatty acids and lipogenesis in the liver.

Evaluation of polyherbal formulation in broilers fed high energy diet: Implications on zootechnical parameters, fat accretion, and serum L-carnitine levels.

Objective: The current broiler trial was planned to assess the effects of Kolin Plus™, a polyherbal formulation (PHF), on performance, protein and fat accretion, and serum L-carnitine (LC) levels in broilers fed a high-energy diet (HED). Materials and Methods: A total of 500 1-day-old Cobb 430 male chicks were assigned to 5 treatment groups consisting of 10 replicates, with 10 birds in each replicate (n = 100). Group G1 was a negative control fed HED, and group G2, a positive control supplemented with synthetic choline chloride (SCC) 1,500 gm/ton in HED. Groups G3, G4, and G5 were supplemented with PHF in HED at 400, 500, and 750 gm/ton feed, respectively (PHF400, PHF500, and PHF750). Results: The PHF produced a dose-dependent numerical improvement in body weight, feed conversion ratio, livability, and the European Production Index. There were no changes in carcass nitrogen and protein accretion, whereas a statistically significant decrease (p < 0.05) in carcass fat and fat accretion was observed in the SCC and PHF groups. Moreover, PHF showed a significant increase in serum LC levels. Conclusion: Kolin Plus™ improves performance parameters akin to SCC by improving fat metabolism and mobilization by enhancing serum LC levels and restoring normal fat accretion.

Anti-diarrhoeal activity of a polyherbal formulation in rats and elucidation of its cellular mechanisms.

OBJECTIVE: The present study was aimed to study anti-diarrhoeal activity of a polyherbal formulation (PHF) in rats and elucidate its mechanism of action. MATERIALS AND METHODS: Anti-diarrhoeal activity of PHF was investigated using castor oil-induced diarrhoea, small intestinal transit and enteropooling models in rats. PHF was tested at 75, 150 and 300 mg/kg rat body weight. Loperamide was used as a reference control for in vivo studies. Anti-secretory action was evaluated against heat labile enterotoxin (from Escherichia coli) induced secretion in rat ileal loop model. The effect of PHF (12.5-100 µg/ml) on cAMP-dependent secretory activity was investigated against forskolin-induced cAMP release in HT-29 cells. RESULTS: PHF demonstrated significant (p≤0.05) anti-diarrhoeal activity by increasing the time for first faecal drop and inhibited diarrhoeal episodes by 43, 58 and 60% at 75, 150 and 300 mg/kg body weight, respectively in a dose-dependent manner. Also, the intestinal transit was inhibited upto 33% and the weight of secretory contents induced by castor oil was significantly reduced by PHF, approximately 29% in enteropooling assay. On the other hand, the intestinal loop instilled with PHF and enterotoxin from E. coli demonstrated 61% inhibition of fluid accumulation as compared to loop instilled with enterotoxin only. In vitro studies indicated that PHF inhibits cAMP release in HT-29 cells corroborating the anti-secretory effects observed in aforesaid studies. CONCLUSION: The results suggest that the PHF possesses anti-diarrhoeal activity, evident through reduced faecal output, decreased intestinal transit and anti-secretory activities.

Effect of dietary supplementation of phytogenic feed additive on performance traits, serum neopterin, and cutaneous basophil hypersensitivity response in heat-induced stress model of broiler chickens.

OBJECTIVE: The trial was aimed at assessing the effect of phytogenic feed additive (PFA), a natural adaptogen, on growth performance, serum neopterin level, and cutaneous basophil hypersensitivity (CBH) response in heat-induced stress model of broilers. MATERIALS AND METHODS: One-day-old Ross 308 chicks (N = 360) were randomly distributed among normal control (NOR), heat-stress control (HSC), and PFA treatment (HSC plus PFA at 200 gm/ton of feed) group. HSC and PFA groups were subjected to heat stress (HS) (32°C-36°C) from 9:00 a.m. to 5:00 p.m. for 35 days. The impact of HS on growth performance, serum neopterin level, and CBH response was assessed. RESULTS: High ambient temperature worsened the performance traits [bodyweight (p < 0.05) and feed conversion ratio] and significantly lowered the serum neopterin level and CBH response in the HSC group when compared to the NOR group. However, supplementation of PFA at 200 gm/ton of feed to birds mitigated the detrimental effects of HS. CONCLUSION: PFA at 200 gm/ton demonstrated the immunomodulatory effect through the restoration of serum neopterin level, CBH response, and growth performance traits in heat-stressed broiler chickens. Thus, PFA can be used as a natural adaptogen to increase the stress resistance and mitigate the negative consequences of various stressors in broiler chickens.

Evaluation of a polyherbal formulation for the management of wet litter in broiler chickens: Implications on performance parameters, cecal moisture level, and footpad lesions.

OBJECTIVE: The study was carried out to develop a wet litter model with magnesium chloride to assess the effectiveness of a polyherbal formulation (PHF) on growth performance, litter and cecal moisture (LCM) level, cecal consistency (CC) score, and footpad lesions (FPLs) score in Ross 308 broiler chickens. MATERIALS AND METHODS: 1,200 one-day-old chicks were assigned into five groups: normal control, negative control [NTC; treated with 1.7% magnesium chloride hexahydrate (MgCl2.6H2O)], and three treatment groups, T1, T2, and T3, where 750, 1,000, and 2,000 gm/ton of PHF, respectively, were supplemented. All the groups were fed a basal diet until day 7. However, the NTC and treatment groups were fed a diet with MgCl2 from days 8 to 42. RESULTS: The addition of MgCl2 for 35 days worsened the growth performance traits in broilers and induced wet litter problems (FPL, high LCM, and poor CC) in the NTC group. However, PHF (750, 1,000, and 2,000 gm/ton) ameliorated the negative effect of a diet with MgCl2 on growth performance and wet litter problems, but a better response with respect to LCM and CC was observed in 2,000 gm/ton of PHF group, followed by that in 1,000 gm/ton of PHF group and 750 gm/ton of PHF group on day 42. CONCLUSION: The wet litter broiler model was developed through excessive feeding of MgCl2, which caused the performance parameters to worsen and the emergence of problems associated with the wet litter. Supplementation with PHF ameliorated these problems and, therefore, it can be used for the management of wet litter in poultry.

Use of in vitro bioassays for assessing botanicals.

In vitro bio assays can play a vital role in evaluating botanicals ranging from comparative screening, interaction studies, bio-activity guided fractionation, biological characterization, assisting in stability studies to studying mechanism of actions. In this review, we present some challenges and common pitfalls of using bioassays for assessing botanicals including guidance to overcome them. The overall objective of this review is to improve the success of botanicals products by incorporating robust bioassays during various stages of research and development.

Perceptron learning rule derived from spike-frequency adaptation and spike-time-dependent plasticity.

It is widely believed that sensory and motor processing in the brain is based on simple computational primitives rooted in cellular and synaptic physiology. However, many gaps remain in our understanding of the connections between neural computations and biophysical properties of neurons. Here, we show that synaptic spike-time-dependent plasticity (STDP) combined with spike-frequency adaptation (SFA) in a single neuron together approximate the well-known perceptron learning rule. Our calculations and integrate-and-fire simulations reveal that delayed inputs to a neuron endowed with STDP and SFA precisely instruct neural responses to earlier arriving inputs. We demonstrate this mechanism on a developmental example of auditory map formation guided by visual inputs, as observed in the external nucleus of the inferior colliculus (ICX) of barn owls. The interplay of SFA and STDP in model ICX neurons precisely transfers the tuning curve from the visual modality onto the auditory modality, demonstrating a useful computation for multimodal and sensory-guided processing.

Bioactive caffeic acid esters from Glycyrrhiza glabra.

Thin layer chromatography bioautography (using DPPH spray reagent) guided fractionation of Glycyrrhiza glabra led to the isolation of two caffeic acid derivative esters, viz. eicosanyl caffeate (1) and docosyl caffeate (2). The two compounds exhibited potent elastase inhibitory activity, with IC(50) values of 0.99 microg mL(-1) and 1.4 microg mL(-1) for 1 and 2, respectively. The compounds also showed moderate antioxidant activity in DPPH and ABTS scavenging assays. The results indicate a possible role of caffeic acid derivatives, in addition to flavonoids in the anti-ulcer properties of G. glabra.

Demonstration of LanthaScreen™ TR-FRET-based nuclear receptor coactivator recruitment assay using PHERAstar, a multi-detection HTS microplate reader.

An attempt was made to demonstrate the possibility of performing LanthaScreen™ TR-FRET based nuclear receptor coactivator recruitment assay using PHERAstar, a multi-detection HTS microplate reader. LanthaScreen™ nuclear receptor coactivator recruitment assay (M/s Invitrogen corporation, USA) was performed using PPAR-gamma receptor preparation in the agonist mode. TR-FRET measurements were done on PHERAstar, a multimode microplate reader (BMG LABTECH, Germany). The Lanthascreen PPAR gamma coactivator recruitment assay was successfully performed in the PHERAstar, multimode microplate reader. This was evidenced by an assay robustness score (Z') of 0.71. The current work demonstrates the suitability of using PHERAstar, a multi-detection HTS microplate reader.for performing LanthaScreen™ TR-FRET based nuclear receptor coactivator recruitment assays.

Geraniol, the putative anthelmintic principle of Cymbopogon martinii.

The essential oil of Cymbopogon martinii (palmrosa) and one of its main constituents geraniol were tested for anthelmintic activity (in vitro) employing the nematode, Caenorhabditis elegans. Both the substances showed potent anthelmintic activity. The ED50 of geraniol was found to be 66.7 mcg/mL, suggesting geraniol as the putative anthelmintic principle of palmrosa oil.

Trypsin inhibitory effect of wedelolactone and demethylwedelolactone.

Wedelolactone (WL) and demethylwedelolactone (DWL) isolated from Eclipta alba were tested in the trypsin inhibition bioassay (in vitro). Both compounds showed potent activity. IC(50) values of WL and DWL were found to be 2.9 and 3.0 microg/mL respectively.

Brine shrimp lethality assay of Bacopa monnieri.

Successive petroleum ether, chloroform, ethanol and water extracts, a saponin rich fraction (SRF) and bacoside A isolated from Bacopa monnieri were tested for brine shrimp lethality. Successive ethanol extracts and SRF showed potent activity. Bacoside A showed the maximum activity with a LC(50) of 38.3 microg/mL. The results confirmed the previous reports of an anticancer effect of Bacopa monnieri and suggest bacoside A as the active constituent.