The regulation of beta-adrenergic receptors (β-ARs) is crucial for maintaining pancreatic function and metabolic balance. However, the rising incidence of metabolic syndrome (MetS) highlights the need to understand how a high-fat simple carbohydrate (HFSC) diet affects β-AR signaling in the pancreas. Despite its potential significance, this aspect remains underexplored. We hypothesized that MetS-induced alterations in β-AR subtypes may disrupt regulatory mechanisms, potentially affecting adenylate cyclase coupling. This study investigated the impact of an HFSC diet on β-AR subtype expression and downstream signaling in pancreatic islets. MetS was induced in C57BL/6J mice through a 150-day HFSC diet. Metabolic changes were assessed through anthropometric parameters, blood glucose levels, lipid profiles, and pancreatic islet morphology. β-AR subtype expression (Adrb1/ADRB1, Adrb2/ADRB2, Adrb3/ADRB3), cAMP levels, and receptor localization were evaluated using quantitative real-time PCR (qRT-PCR), Western blotting, and immunofluorescence. The HFSC diet led to metabolic disturbances and pancreatic inflammation. Adrb1 mRNA expression was elevated (p < 0.001), but β1-AR localization did not show a corresponding increase. Both β2-AR transcription and translation were downregulated (p < 0.001). Although β3-AR protein levels were markedly increased (p < 0.001), cAMP levels were significantly reduced (p < 0.01). These findings indicate significant β-AR dysregulation under metabolic stress. Altered β1-AR and β2-AR expression, along with increased but functionally insufficient β3-AR signaling, suggest impaired adrenergic regulation in pancreatic islets during MetS. This study contributes to understanding MetS pathophysiology and highlights the relevance of β-AR signaling as a potential therapeutic target.

Background:: Investigating the structural attributes of the murine beta3-adrenergic receptor (β3-AR) is imperative for comprehending metabolic regulation, given its close resemblance to the human β3-AR. This receptor holds promise as a target for novel drug development against obesity and diabetes. Despite its potential, the absence of knowledge regarding the structure of murine β3- AR hampers a comprehensive understanding of its functionality. Objective:: Our study aimed to model the three-dimensional (3D) structure of murine β3-AR through various molecular structure prediction and simulation techniques, thus addressing the existing gap in structural information. Methods:: Employing diverse structure prediction programs, we refined the predicted structure of murine β3-AR. Primary sequence analysis offered insights into charge distribution, stability, and hydrophobic properties. The binding sites were identified in the modeled structure. Molecular Dynamics (MD) simulation provided the structural stability and dynamic behavior of the predicted β3- AR structure. Results:: The β3-AR protein exhibited specific characteristics, including a pI of 9.57, an aliphatic index of 98.35, a GRAVY score of 0.289, and the presence of conserved motifs and disulfide linkages. Utilizing the programs such as Phyre2, Swissmodel, I-Tasser, and AlphaFold2, we generated a 3D model of murine β3-AR. Subsequent refinement using ModRefiner revealed a structure comprising 13 helices, 2 strands, and 21 turns. The Ramachandran plot indicated favorable regions for 93.2% of residues, with minimal deviations. A 50 ns MD simulation demonstrated the consistent stability and integrity of the β3-AR protein. The top three binding pockets were identified based on varying areas and volumes. Dynamic behavior within residues Ser 252 and Arg 253 was observed, indicating flexibility in conformation. This study marks the first-ever exploration, offering initial structural insights into murine β3-AR. Conclusion:: This study underscores the critical role of computational approaches in predicting the 3D structure of β3-AR. We derived a refined model by employing diverse prediction techniques, elucidating key features. The findings emphasize the significance of this methodology in comprehending the structural foundation of β3-AR, providing valuable insights for targeted medication development against conditions such as obesity and diabetes.

In India, approximately 80,000 tonnes of flower waste are produced every year. The increase in floral waste has driven attention to utilise it in various fields. Flowers, with their intricate properties, are applicable in the dye, paper, incense, perfumery, and pharmaceutical industries. It can also be used for vermicomposting. The pigments extracted from temple flowers are known to show antibacterial and antifungal activity and are finding their application as anti-cancerous agents. This paper comprises an overview of the major floral pigments such as carotenoid, flavonoids and xanthophyll, which are found in temple flowers and their extraction procedures using conventional methods such as agitation, centrifugation, Soxhlet extraction and non-conventional methods such as enzymatic extraction, microwave-assisted extraction, pressurised liquid extraction and ultrasound-assisted extraction. This is followed by a brief discussion of the cytotoxic effect in vitro using cell lines such as MCF-7, HeLa, and DU145.

Microbial Exopolysaccharides: Natural Modulators of Dairy ProductsPallavi Jaiswal, Rohit Sharma, Bhagwan Singh Sanodiya, Prakash Singh Bisen

The impact of bacterial microbiome metabolites on human health, most particularly on the prevention and treatment of cancer, has gained significant interest in recent times. The study focused on the isolation of bacterial flora associated with the ovaries of 8-week-old mice and examined its ability to produce metabolites with potential antibacterial, antioxidant, and anticancer properties. The isolates were identified based on biochemical and 16S rRNA sequencing. The ethyl acetate extract from isolates was screened for antimicrobial and antioxidant capacity. The 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay was used to evaluate the cytotoxicity of the selected isolate against the ovarian carcinoma cell lines SKOV-3 and PA-1. The ethyl acetate extract from Bacillus velezensis OM03 exhibited significant antibacterial activity against all the tested bacteria, with a MIC value ranging from 50 to 100 µg/mL. Furthermore, the extracts demonstrated hydrogen peroxide and 2, 2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activities at rates of 88.50% and 87.78%, respectively. The extracts displayed substantial concentration-dependent antiproliferative/cytotoxic activities against SKOV-3 and PA-1 cell lines after 24 and 48 h of treatments, respectively. Further chemical analysis of the extract using HR-LCMS revealed the presence of bioactive compounds such as myriocin, 2,3-diethylpyrazine, dihydrodeoxystreptomycin, cyclo (L-Phe-L-Pro), C16 sphinganine, and other twenty-nine compounds that have been formerly reported and are accountable for the targeted activities. Bacillus velezensis OM03 may be further investigated for the creation of novel therapeutics, particularly for the treatment of ovarian cancer.

Searchable abstracts of presentations at key conferences in endocrinology ISSN 1470-3947 (print) | ISSN 1479-6848 (online)

Introduction and Aim: The health advantages, enhanced organoleptic features, and global economic significance of probiotic-enriched food items have motivated researchers to discover novel probiotics. In light of this, a study was performed to assess the probiotic potential of five Bacillus strains isolated from the ovaries of 8-week-old mice in a previous study. Materials and Methods: The five Bacillus strains were appraised for their probiotic potentials, such as tolerance to acidic pH, bile salt, and phenol. The haemolytic behaviour, antibiotic sensitivity characteristics, and capacity to produce amylase, protease and L-asparaginase were also assessed. Results: All the isolates exhibited high survival in simulated gastric juice conditions such as pepsin containing acidic pH environments, up to 1% bile, and 0.5% phenol. Bacillus cereus OM02 and Bacillus velezensis OM03 exhibited the highest tolerance, with 50 % of survival at pH 2.5 and 1% bile salt-containing media. All the isolates were found to be amylase and protease producers and non-beta-haemolytic, however, OM02 and OM04 were beta-haemolytic. Extracts from the strain OM03 also exhibited dosage-dependent ferric-reducing antioxidant power. Except for OM04, which is resistant to ampicillin and sulphatriad, and OM01 and OM02, which are sulphatriad-resistant, all isolates were susceptible to most antibiotics. This is the first study on Bacillus species from Swiss albino mouse ovarian to evaluate probiotic potential. Conclusion: These findings suggest that Bacillus velezensis OM03, Bacillus aerius OM01, and Bacillus subtilis OM05 are suitable probiotic candidates and may be used in both animal and human formulations to improve consumer health.

Introduction and Aim: This study reports the cytotoxic potential of L-Asparaginase isolated from the fungus Scopulariopsis brevicaulis. Materials and Methods: Extracellular L- Asparaginase was isolated from Scopulariopsis brevicaulis and purified by ammonium sulfate precipitation, followed by dialysis, ion exchange and gel filtration chromatography. Varying concentrations (31.25, 62.5, 125, 250, 500 µg/ml) of purified L-Asparaginase was tested on MCF7, HeLa, HepG2 and 3T3L1cell lines by MTT assay. Curcumin was maintained as a positive control. Results: Minimum inhibition of 23.57% was observed at an enzyme concentration of 31.25 µg/ml and maximum inhibition (66.41%) was observed at 500 µg/ml against MCF7 cell line. Minimum inhibition of 2.87% was observed at an enzyme concentration 31.25 µg/ml and maximum inhibition (58.49%) was observed at 500 µg/ml against HeLa cell line. Minimum inhibition of 4.58% was shown at an enzyme concentration of 31.25 µg/ml and maximum inhibition (46.14 %) was observed at 500 µg/ml against HepG2 cell line. Minimum inhibition of 1.4% was shown by enzyme concentration 31.25 µg/ml and maximum inhibition (50.9%) was observed at 500 µg/ml against 3T3L1 cell line. Conclusion: We report for the first time the cytotoxic potential of L-Asparaginase from Scopulariopsis brevicaulis.

This study reports the cytotoxic potential of L-Asparaginase isolated from Bacillus sp. Bacillus sp. was isolated from local soil/water samples and identified by rapid plate assay and further confirmed by phenotypic characterization. Extracellular L- Asparaginase was isolated from broth culture of Bacillus sp. and purified by ammonium sulfate precipitation, followed by dialysis and ion exchange and gel filtration chromatography techniques. The purified enzyme was used to study the in vitro cytotoxic potential. Varying concentrations (31.25, 62.5, 125, 250, 500 µg/ml) of purified L-Asparaginase was tested on MCF7, HeLa, HepG2 and 3T3L1cell lines by MTT assay. Curcumin was maintained as a positive control. The results revealed that the enzyme showed a significant cytotoxic activity and a dose dependent effect. Minimum inhibition of (19.44%) was observed at an enzyme concentration of 31.25 µg/ml and maximum inhibition (71.14%) was observed at 500ug/ml against MCF7 cell line. Minimum inhibition of (10.04%) was observed at an enzyme concentration 31.25 µg/ml and maximum inhibition (68.92%) was observed at 500 µg/ml against HeLa cell line. Minimum inhibition of (7.45%) was shown at an enzyme concentration of 31.25 µg/ml and maximum inhibition (68.28 %) was observed at 500 µg/ml against HepG2 cell line. Minimum inhibition of (4.4%) was shown by enzyme concentration 31.25 µg/ml and maximum inhibition (47.4%) was observed at 500 µg/ml against 3T3L1 cell line. Curcumin at 5µM concentration showed an inhibition of 48.13% against MCF-7 cells, 54.42% against HeLa cells, 64.94% against HepG2 cells and 44.5% against 3T3L1 cell lines.

Given the association between subclinical hypothyroidism and metabolic syndrome, we wanted to explore if high-fat, simple-carbohydrate (HFSC) diet affects hypothalamus-pituitary-thyroid axis. One-month-old male C57BL/6J mice were fed with control (C) and HFSC (T) feed (n = 18 each), respectively, for 5 months. There was a significant increase in triiodothyronine in the T group (13.5%) compared with the age-matched C group by the fifth month. Thyroid-stimulating hormone was significantly higher (1 month: 1.9-fold; 3 months: 2.66-fold; 5 months: 3.5-fold) from the first to fifth months in the T group compared with age-matched C group. Thyrotropin-releasing hormone (TRH) gene expression showed significant decrease (1 month: 83.2%; 5 months: 40.7%) in the T group compared with the age-matched C group. TRHR1 showed significant decrease in the T group compared with the age-matched C group throughout the study (1 month: 82.8%; 3 months: 45.7%; 5 months: 75.2%). However, TRHR2 showed dynamic change during the study. Initially there was significant (1 month: 0.104-fold) downregulation, followed by significant upregulation (3 months: 3.6-fold) and downregulation (0.73-fold) by the fifth month in the T group compared with the age-matched C group. There was marked depletion of functional follicular cells and colloid substance in the thyroid glands of the T group by the fifth month compared with the C group. Leptin receptors ObRa (1 month: 48.25%; 5 months: 88%) and ObRb (1 month: 46.9%; 5 months: 63.3%) were significantly downregulated in the T group compared with the age-matched C group in the first and fifth months of feeding the respective diets. The expression of p-STAT3, a transcription factor known to have a role in energy balance, intermediate metabolism, and leptin signalling was seen to decrease significantly (6.25-fold) in the hypothalamus of the T group compared with the age-matched C group. In conclusion, HFSC feed disrupts the hypothalamus-pituitary-thyroid axis in male C57BL/6J mice.

Objectives: To study the effect of specially formulated high-fat simple carbohydrate diet (HFSC) on the serotonin metabolic pathway in male C57BL/6J mice. Methods: Previous studies from our laboratory have shown that specially formulated HFSC induces metabolic syndrome in C57BL/6J mice. In the present investigation, 5-hydroxytryptophan, serotonin and 5-hydroxyindoleacetic acid were analyzed in two brain regions (hypothalamus, corpus striatum), urine and plasma of HFSC-fed mice on a monthly basis up to 5 months using high-performance liquid chromatography fitted with electrochemical detector. The data were analyzed using Graph pad Prism v7.3 by two-way ANOVA and post hoc Tukey’s test (to assess the effect of time on the serotonergic metabolic pathway). Results: HFSC feed was observed to lower the hypothalamic serotonergic tone as compared to the age-matched control-fed C57BL/6J mice. Although the hypothalamic serotonergic tone was unaltered over time due to consumption of diet per se, hypothalamic 5-HTP levels were observed to be lower on consumption of HFSC feed over duration of 5 months as compared to 1st month of consumption of HFSC feed. The striatal 5-HTP levels were lowered in the HFSC-fed mice after 4 months of feeding as compared to the age-matched control-fed mice. The striatal 5-HTP levels were also lower in both control and HFSC-fed mice due to consumption of the respective diet over a duration of 5 months. Increased plasma 5-HTP levels were observed due to consumption of HFSC feed over duration of 5 months in the HFSC-fed group. However, higher breakdown of serotonin was observed in both the plasma and urine of HFSC-fed C57BL/6J mice as per the turnover studies. Discussion: The central and peripheral serotonergic pathway is affected differentially by both the type of diet consumed and the duration for which the diet is consumed. The hypothalamic, striatal and plasma serotonergic pathway were altered both by the type of feed consumed and the duration of feeding. The urine serotonergic pathway was affected by mainly the duration for which a particular diet was consumed. These findings may have implications in the feeding behavior, cognitive decline and depression associated with metabolic syndrome patients.

Metabolic syndrome (MetS) refers to clustering of lipid abnormalities and insulin resistance in an individual. Its increasing prevalence in children and adolescents can be curbed by generating pertinent awareness among students at educational institutions regarding importance of BMI, nutritional content of food, food habits etc. Its prevalence is reported to vary according to racial and ethnic groups urging the need for focused studies by educational institutions on the risk factors relevant to the Indian population. Universities can also aim at introducing information regarding lifestyle interventions, plan outreach programs to ingrain the information not only in the student population but also the society as a whole. In this book chapter, we propose to review literature which will help in identifying areas that need immediate attention, possible implementation strategies in the education system along with the efforts of our lab towards understanding the etiology of this epidemic.

INTRODUCTION:Very few systematic studies are done during the onset and progression of metabolic syndrome in suitable animal models. In this paper we present the effect of High-Fat Simple Carbohydrate (HFSC) feed on the metabolic hormones in C57BL/6J mice to understand the sequence of events leading to impairment of glucose homeostasis.MATERIAL AND METHODS:One-month-old male C57BL/6J mice were fed with control (C group) and HFSC (T group) feed (n = 30 each) respectively for five months. The glucose tolerance was studied by Oral Glucose Tolerance Test (OGTT) whereas serum insulin and leptin were quantified using ELISA kits, and serum cortisol was quantified using CLIA kits.RESULTS:Insulin resistance index and HOMA-IR levels were higher in the mice of group T as compared to age-matched mice of group C within one month and significantly higher after and five months of feeding. The total area under the glucose tolerance test curve (AUC) and the insulin curve (AUC ins) was found to significantly increase in the mice of T group as compared to the mice of C group as early as two months of feeding and was elevated after 5 months post feeding. Comparison of the Matsuda index revealed that pancreatic beta cell function was significantly lower in mice of T group as compared to mice of C group by five months of feeding. Leptin levels fluctuated during the 1st-4th month and by the 5th month significant hyperleptinaemia was detected. There was no significant change in cortisol levels in mice of group T as compared to mice of group C after five months of feeding.CONCLUSIONS:HFSC feed induces insulin resistance by the first month and progressively impairs glucose tolerance, resulting in hyperleptinaemia by the fifth month in male C57BL/6J mice. (Endokrynol Pol 2016; 67 (6): 592-598).

Purpose: Many diet based studies on metabolic syndrome lack an in-depth study of feed nutrient profile post preparation. We analyzed the nutritional profile of formulated High Fat Simple Carbohydrate (HFSC) feed and studied its effect on target organs. Methods: Control feed and HFSC feed were analyzed for macro and micronutrient profile and fed to one month male C57BL/6J mice for 5 months during which feed intake, energy intake and feed efficiency was monitored. The effects of feeds were preliminarily studied on kidney, liver, adrenal gland and pancreas by histological staining. Results: The HFSC feed had significantly lower carbohydrate, moisture, crude fiber, ash content, mineral, vitamin content, higher ω-6/ω-3 fatty acid ratio as compared to control feed. The HFSC fed mice had higher food intake; energy intake; feed efficiency; lymphocyte infiltration in the liver; hypertrophy of the kidney, adrenal medulla and degenerated islets of pancreas as compared to the control mice after 5 months of feeding. Conclusion: Food with a higher ω-6/ω-3 fatty acid ratio and simple carbohydrate leads to deterioration of structural integrity of vital organs.

High fat simple carbohydrate fed C57BL/6J mice, a model for metabolic syndrome have shown significant disruption of feedback control between TSH of the pituitary and T3 hormone of the thyroid gland. In order to understand the exact mechanism involved, it is important to assess the level of the hypothalamic TRH. qRT-PCR is a method of choice when tissue sample is less. In the present study, we have designed C57BL/6J mice specific primers for TRH.

A previous study from our lab has shown derailment of adrenergic receptors during onset and progression of Metabolic Syndrome. Hence, gene expression study was proposed to understand the subtype of adrenergic receptors. Primer designing is the first and foremost step in the gene expression studies. In this work, suitable primers were designed for SYBR green chemistry and validated using bioinformatics tools to study the adrenergic receptor subtypes involved during onset and progression of Metabolic Syndrome.

BACKGROUND:Previous studies have shown disturbances in an individual monoamine pathway but have not studied metabolic pathways at the onset and progression of metabolic syndrome (MetS). Aims, Settings, and Design: The aim of this study was to investigate the effect of high-fat simple carbohydrate (HFSC) diet on central (hypothalamic) and peripheral (plasma and urine) monoamine metabolic pathways during the development of metabolic syndrome in C57BL/6J mice. MATERIALS AND METHODS:Monoamines were analyzed in the 1st, 2nd, 3rd, 4th, and 5th month after feeding mice the HFSC diet or the control diet using the high performance liquid chromatography (HPLC) system (Shimadzu, Japan). Data was statistically analyzed (by Student's t-test) using Graph Pad Instat Version 3.1. Post statistical analysis, Bonferroni correction was applied to the results of 2nd, 3rd, 4th, and 5th month in order to calculate the correct error in the study. RESULTS:Significantly lower hypothalamic, plasma, and urine dopamine, and higher hypothalamic and plasma levels of norepinephrine and normetanephrine levels were observed in the HFSC diet fed C57BL/6J mice as compared to the control diet fed C57BL/6J mice after 5 months of feeding. No consistent changes were observed in other brain regions. The turnover ratio indicated that the lower dopamine levels in the HFSC diet fed C57BL/6J mice was due to the increased formation of norepinephrine and homovanillic acid. CONCLUSION:HFSC diet impairs the central and peripheral dopaminergic and noradrenergic pathways in mice as evidenced by the disturbances in their hypothalamic, plasma, and urine levels and this might be one of the early factors contributing towards the development of the MetS.

Background: Consumption of nuts particularly walnuts has been widely recommended for lowering the risk of coronary heart disease; however, very few studies have evaluated the potential unfavourable effects that Walnut seeds might have in vivo.Objective: To investigate the long-term effects of Juglans regia seed supplementation in specially formulated feed when fed to developed mice model.Method: C57BL/6J male mice were divided into four groups, labelled, C (control), T (High Fat Simple Carbohydrate-HFSC), Cw (control + walnut) and Tw (HFSC+ walnut) based on diet. Four mice from each group were sacrificed at the end of the first and fifth month respectively. Blood samples were collected every month. Bleeding time, blood platelet number and morphology were studied. Histopathological analysis of heart and aorta were also performed.Results: Groups Cw and Tw showed a significant increase in platelet count; however, platelet activation was not detected. There was a trend to an increase in bleeding time in group Cw. Expanded lumen diameter of the aorta was observed in Cw and Tw mice initially, however, it was found to constrict by the end of the study. A large amount of peripheral fat deposition in the aortas of Cw and Tw was observed, which increased through the course of the experiment.Conclusion: Kashmiri walnut seed supplementation was found to reduce the signs in shape change of platelets as well as increase platelet number with a trend to an increased bleeding time. This observation could be of medical interest. Histopathological analysis of heart and aorta showed lipid sequestration which merits further investigations.

Metabolic syndrome represents a combination of cardiovascular risk determinants and lipid abnormalities. The syndrome is associated with a five-fold higher risk of developing type-II diabetes and two to threefold higher risk of cardiovascular disease. The etiology of the syndrome at the molecular level is not yet understood. Therefore, there is a need to study the syndrome in an animal model. In the present study, the effect of high fat simple carbohydrate (HFSC) diet on male Wistar rats was investigated. The rats were fed HFSC diet for a period of five months. Anthropometric parameters were measured and serum triglycerides, total cholesterol, HDL and LDL were analysed. Glucose intolerance was studied using oral glucose tolerance test. The formulated (HFSC) diet succeeded in developing an animal model for metabolic syndrome within 17 weeks characterized by obesity, hypertriglyceridemia, impaired glucose tolerance, fatty liver and abnormal cardiac histology.

Metabolic syndrome is a multiplex risk factor arising from abdominal obesity, insulin resistance and dyslipidemia.It increases the risk for cardiovascular complications, fatty liver disease and cognitive dysfunction.The prevalence is rampantly increasing among all age groups worldwide due to nutrition excess, physical inactivity and increased stress.Despite its epidemiological nature, treatment is limited to lifestyle changes and lifelong medical intervention.This may be due to incomplete knowledge of the etiology of the syndrome.The present study is aimed at developing an animal model using male C57BL/6J mice and studying the biochemical profile during the onset and progression of the syndrome.Test group was fed with High Fat Simple Carbohydrate (HFSC) diet and the Control group was fed with standard diet for a period of 5 months.Body weight and biochemical profile was analyzed on a monthly basis.Oral Glucose tolerance test (OGTT) was performed following a glucose load of 1g/kg body weight at 143 rd day.The syndrome was successfully induced in male C57BL/6J mice within a period of 5 months with perturbed lipid profile and insulin resistance.