Interplay of precision therapeutics and MD study: Calocybe indica's potentials against cervical cancer and its interaction with VEGF via octadecanoic acid.

The evolving landscape of personalized medicine necessitates a shift from traditional therapeutic interventions towards precision-driven approaches. Embracing this paradigm, our research probes the therapeutic efficacy of the aqueous crude extract (ACE) of Calocybe indica in cervical cancer treatment, merging botanical insights with advanced molecular research. We observed that ACE exerts significant influences on nuclear morphology and cell cycle modulation, further inducing early apoptosis and showcasing prebiotic attributes. Characterization of ACE have identified several phytochemicals including significant presence of octadeconoic acid. Simultaneously, utilizing advanced Molecular Dynamics (MD) simulations, we deciphered the intricate molecular interactions between Vascular Endothelial Growth Factor (VEGF) and Octadecanoic acid to establish C.indica's role as an anticancer agent. Our study delineates Octadecanoic acid's potential as a robust binding partner for VEGF, with comprehensive analyses from RMSD and RMSF profiles highlighting the stability and adaptability of the protein-ligand interactions. Further in-depth thermodynamic explorations via MM-GBSA calculations reveal the binding landscape of the VEGF-Octadecanoic acid complex. Emerging therapeutic innovations, encompassing proteolysis-targeting chimeras (PROTACs) and avant-garde nanocarriers, are discussed in the context of their synergy with compounds like Calocybe indica P&C. This convergence underscores the profound therapeutic potential awaiting clinical exploration. This study offers a holistic perspective on the promising therapeutic avenues facilitated by C. indica against cervical cancer, intricately woven with advanced molecular interactions and the prospective integration of precision therapeutics in modern oncology.

Expression profile of different classes of proteases in milk derived somatic cells across different lactation stages of indigenous cows (Bos indicus) and riverine buffaloes (Bubalus bubalis).

Proteases play a significant role in milk and its products by affecting flavor, texture and longevity. The expression of endogenous proteases varies across different stages of lactation. The study was conducted to understand the transcriptional pattern of different classes of protease-pathways associated genes (CTSB, CTSD, CTSH, CTSL, CTSK, CTSS, CTSZ, PLAU, PLAT) and potential protease inhibitors (SERPIN E2 and SERPIN F2) in 40 milk somatic cells (MSC) samples isolated during early, peak, mid and late lactation stages of Sahiwal cows and Murrah buffaloes - the two most important dairy breeds of India. In Sahiwal cows, except CTSK and PLAU, the expression of other proteases class was not affected significantly (p > 0.05) across lactation stages. However, in Murrah buffaloes, the expression of different proteases increased as the lactation progressed. Most of the proteases showed lower expression during early and peak lactation stages while their expression tends to increase during mid to late lactation stages. The overall trend was somewhat similar in both the dairy species albeit the level of expression was higher in buffalo MSC as compared to cow MSC. The study has provided valuable information on expression kinetics of different proteases in milk somatic cells of two major dairy breeds of India.

Impact of a behaviourally focused nutrition education intervention on attitudes and practices related to eating habits and activity levels in Indian adolescents.

OBJECTIVE: To evaluate the effectiveness of a behaviourally focused nutrition education (NE) intervention based on the Health Belief Model (HBM) to improve knowledge, attitudes and practices (KAP) related to eating habits and activity levels in 10­12-year-old adolescents in Mumbai, India. DESIGN: School-based cluster randomised controlled trial. The experimental group (EG) received weekly NE and three parent sessions over 12 weeks; no sessions were conducted for the control group (CG). The theoretical framework of HBM and focus group discussion results guided the development of behaviour change communication strategies and NE aids. KAP were measured using a validated survey instrument, administered at baseline and endline in EG and CG. Paired and independent t tests determined within-group and between-group changes in pre­post scores. SETTING: Two aided and two private schools that were randomly allocated to either an EG or CG. PARTICIPANTS: Adolescent boys and girls (n 498; EG n 292 and CG n 206). RESULTS: EG reported improvements in mean knowledge (39·3%), attitude (7·3 %), diet (9·6 %) and activity practice (9·4%) scores from pre to post intervention. No significant changes were observed in CG. Significant improvements in scores associated with perceived benefits, barriers and self-efficacy, breakfast and vegetable consumption, and moderate-to-vigorous activities were observed in EG. CONCLUSIONS: Integrating NE into the academic curriculum and adopting evidence-based lessons that entail targeted information delivery and participatory activities can improve knowledge, foster right attitudes and facilitate better eating and activity-related practices in Indian adolescents.

Independent and combined influences of physical activity, screen time, and sleep quality on adiposity indicators in Indian adolescents.

BACKGROUND: Inadequate physical activity (PA), excess screen time (ST), and sub-optimal sleep quality tend to co-occur during adolescence. Yet, little is known about the associations of these behaviors as a cluster with adiposity indicators in Indian adolescents. This study aimed to evaluate the independent and combined influences of PA, ST, and sleep quality on body mass index (BMI) and waist to height ratio (WHtR) in 10-15 years old adolescents in Mumbai, India. A secondary aim was to explore if these influences vary between sexes. METHODS: Cross-sectional study. Adolescents (n = 772, mean age 13.2 (1.4) years) reported frequency and duration of moderate to vigorous PA (MVPA) and time spent using screens on a previously validated instrument. Sleep quality was estimated using the Pittsburg Sleep Quality Index (PSQI). Weight, height, and waist circumference were measured. Mixed effect logistic regression analyses were performed to explore associations between adiposity indicators (BMI z scores > +1SD and WHtR > 0.5) and unhealthy behaviors (PA < 60 min/d, ST > 120 min/d and PSQI scores > 5), stratified by sex. RESULTS: The combined prevalence of overweight and obesity was 38.3%. Overall, 62.0 and 85.0% reported MVPA< 60 min/d and ST > 120 min/d respectively. Girls reported higher ST (218.21 (69.01) min/d) as compared to boys (165.3 (101.22) min/d, p < 0.001). Clustering of low PA and excess ST was observed in 69.2% and of all three unhealthy behaviors in 18.8%. Among girls, MVPA < 60 min/d (OR = 1.78, 95% CI 1.54-1.92, p < 0.001) and PSQI scores > 5 (OR = 2.01, 95% CI 1.78-2.25, p < 0.001) predicted increased BMI. The odds of overweight/obesity were 2.10 times higher in boys reporting low PA and 4.13 times higher in those with low PA+ ST > 120 min/d. Clustering of all three unhealthy behaviors increased prevalence of obesity in both sexes. CONCLUSIONS: The results indicated a co-existence of multiple unhealthy lifestyle factors of obesity and that clustering of these behaviors can further aggravate obesity risk as compared to their independent effects. Integrated interventions that leverage the cumulative benefits of being active, less sedentary and sufficient sleep are warranted to facilitate greater improvements in obesity risk behaviors.

Development and validation of a questionnaire measuring knowledge, attitudes, and practices (KAP) to healthy eating and activity patterns in school children (HEAPS).

BACKGROUND: Development of culturally appropriate and psychometrically sound instruments that measure knowledge and health behaviors of children will help to inform appropriate interventions. AIM: To develop and test the validity and reliability of a questionnaire measuring knowledge, attitudes, and practices to healthy eating and activity patterns in school children in India. METHODS: Review of literature, focus-group discussions, and theoretical constructs of the Health Belief Model guided the development of an item pool. Face and content validity were assessed by children and a panel of experts and the item content validity, item difficulty, and discrimination indices were calculated. Construct validity was determined using the principal axis method of exploratory factor analysis among a cross-sectional sample of children (n=252). Internal consistency (Cronbach α values >0.7) and test-retest reliability (intraclass correlation coefficient values >0.75) were estimated. RESULTS: Item content validity index for clarity and relevance were satisfactory (>0.80) and internal consistency for knowledge (Kuder-Richardson 20 = 0.832), attitude (Cronbach's α = 0.912), and practice items (Cronbach's α = 0.769) were good. Four factors (children's eating habits, family dietary practices, and consumption of healthy and unhealthy foods) and two factors (moderate to vigorous activities and sedentary activities) explained 67.7% and 48.2% of the total variance in practice items. Intraclass correlation coefficient estimates ranged from good to excellent (0.72-0.99). CONCLUSIONS: The results of the validity and reliability of the 84-item knowledge, attitudes, and practices to healthy eating and activity patterns in schoolchildren questionnaire were promising. The detailed description of the methodology employed may prove useful to researchers conducting similar studies in children.

Responses of diversity, productivity, and stability to the nitrogen input in a tropical grassland.

Atmospheric nitrogen (N) deposition is a matter of serious concern for the structure and functioning of global ecosystems, but the effect of N application of species diversity (D), primary productivity (P), and stability (S) of tropical grassland ecosystems is not known. The present study reports the effects of different levels of N application on species composition, and the D, P, S, and their relationships in a tropical grassland. Within the experimental grassland, 72 1 × 1 m plots with 6 N-input levels and with 12 replicates, were established in 2013. For 3 yr, different doses of urea as a source of N were applied to the plots. Data on individuals and biomass of each species were recorded and statistically analyzed. The study revealed that the N applied caused variations in species composition, D, P, and S. Below the 90 kg N dose, D was positively related to P and S while, above this level, the relations were negative due to N-induced responses of species and functional group composition as well as biomass distribution among them. The optimum applied N levels for maximum D (50-60 kg N), P (120 kg N), and a positive relationship of S with D (up to 90 kg N treatment) suggested that the 90-kg N dose could be the maximum dose of N that the grassland can tolerate. Hence, N application should not exceed the 90-kg level for sustainability of the structure and functioning of tropical grassland ecosystems.

Characterizing binding sites of heat responsive microRNAs and their expression pattern in heat stressed PBMCs of native cattle, exotic cattle and riverine buffaloes.

It is generally believed that due to evolutionary differences and adaptation to tropical conditions, Indian native cattle has superior heat tolerant ability than Bos taurus cattle. In the present study, 3'-UTR of two most important heat responsive genes i.e., heat shock protein 70.1 (HSP70.1) and heat shock factor- 1 (HSF-1) were sequence characterized in different breeds of Indian native cattle to identify the variations and miRNA binding sites. In addition, the impact of heat stress was assessed in a total of 57 PBMCs samples of native Sahiwal cows (Bos indicus), exotic Holstein cows (Bos taurus) and Murrah buffaloes (Bubalus bubalis) using various cellular parameters like cell viability, cytotoxicity and apoptosis. Further, expression profile of 12 heat responsive miRNAs were also evaluated in unstressed and stressed PBMCs to understand post transcriptional changes in native cows, exotic cows and Murrah buffaloes. The sequence data showed 3'-UTR of HSP70.1 gene of Indian cattle to be exactly similar to Bos taurus with no miRNA binding site. Whereas, sequencing of 3'-UTR of HSF-1 gene revealed 3 SNPs at positions G1762T; C1811T and C1983T with 7 well conserved miRNA binding sites. The impact of heat stress on various cellular parameters in terms of cell viability, cytotoxicity and apoptosis was highest in PBMCs of Holstein cows followed by Murrah buffaloes and Sahiwal cows. Further, in contrast to Holstein Frisian cows and Murrah buffaloes, the expression pattern of 12 heat responsive miRNAs, in heat stressed PBMCs of Sahiwal cows were quite distinct. There was a significant (p < 0.05) induction in expression of most of the miRNAs after heat stress in PBMCs of Sahiwal cows followed by a rapid decline. The distinct cellular response and pattern of miRNA expression across cattle types and buffaloes might be influencing their PBMCs tolerance level to heat stress.

Nickel Selenide Nanoparticles as a Cheap Alternative for Pt-Counter Electrode in Dye-Sensitized Solar Cells.

Colloidal nickel selenide nanocrystals (NCs) of two different compositions i.e., Ni3Se2 and Ni3Se4, where one is Ni-rich while another is Se-rich are synthesized using the hot injection method by merely changing the injection and growth temperature while keeping the rest of the reaction conditions like solvent, ligands, amount of precursors and reaction time identical. These nanocrystals exhibit electrocatalytic activity in the reduction of triiodide (I-3) to iodide (3I-), therefore employed as counter electrodes (CE) in dye-sensitized solar cells (DSCs). The DSC based on nickel selenides displays an efficiency of 6.4%, comparable to the Pt-based cells prepared by us.

Comparative Evaluation of Continuous Veno-venous Hemodiafiltration and Continuous Arterio-Venous Hemodiafiltration in Patients of Hepatic Failure and / or Hepatorenal Syndrome.

OBJECTIVES: Continuous renal replacement therapies (CRRT) are the most favoured form of renal replacement therapies (RRT) in patients of decompensated liver cirrhosis and hepatorenal syndrome (HRS). The role of CRRT has been limited only to acute kidney injury and HRS in prior studies. We therefore aimed to evaluate the role of two different modes of CRRT- CVVHDF and CAVHDF in patients of hepatic failure and / or hepatorenal syndrome in reducing hyperbilirubinemia, uremia and fluid overload. METHODS: 30 critically ill patients of hepatic failure and /or HRS were randomly divided into two groups of 15 cases each. Group A patients received continuous veno-venous hemodiafiltration (CVVHDF), whereas group B patients underwent continuous arterio-venous hemodiafiltration (CAVHDF). The inclusion criteria were hepatic failure and / or hepatorenal syndrome (HRS) with hyperbilirubinemia and fluid overload in hemodynamically unstable patients, who were unfit for conventional hemodialysis. RESULTS: Despite hemodynamic fragility of the subjects, both the procedures were effective in achieving biochemical and clinical improvements. There was a significant fall in blood urea, serum creatinine and serum bilirubin at the end of procedures. After mean 27.32 h of CVVHDF and 27.02 h of CAVHDF, blood urea decreased to 39.54 ± 28.6 mg/dl and 45.11 ± 31.9 mg/dl in respective groups. Serum bilirubin decreased to 7.01 ± 6.4 mg/dl and 3.21 ± 1.99 mg/dl in group A and B. All the patients had gradual and steady improvement in pH and bicarbonate concentration towards normal. Urea clearance was 24.98 ± 1.09 ml/min and 22.72 ± 1.58 ml/min respectively in the two groups, whereas bilirubin clearance was 27.77 ± 1.38 ml/min and 28.74 ± 0.3 ml/min in group A and B respectively. Ultrafiltration rate had mean value of 141.66 ± 22.33 ml/h in group A and 134.26 ± 38.71 ml/h in group B. Both the modes of CRRT were well tolerated without any new episodes of hypotension secondary to the procedures and requirement of inotropes didn't change significantly. Symptomatic relief and improvement in clinical and biochemical parameters were observed in all the cases. There were no significant differences between the results of two groups. Complication rate was less and survival was 30%. CONCLUSION: Continuous hemodiafiltration is probably the best available modality of CRRT to treat hemodynamically unstable and critically ill patients of hepatic failure and/ or hepatorenal syndrome and it should be advocated more frequently.

Genotyping-by-sequencing through transcriptomics: implementation in a range of crop species with varying reproductive habits and ploidy levels.

The application of genomics in crops has the ability to significantly improve genetic gain for agriculture. Many marker-dense tools have been developed, but few have seen broad adoption in plant genomics due to issues of significant variations of genome size, levels of ploidy, single nucleotide polymorphism (SNP) frequency and reproductive habit. When combined with limited breeding activities, small research communities and scant sequence resources, the suitability of popular systems is often suboptimal and routinely fails to effectively balance cost-effectiveness and sample throughput. Genotyping-by-sequencing (GBS) encompasses a range of protocols including resequencing of the transcriptome. This study describes a skim GBS-transcriptomics (GBS-t) approach developed to be broadly applicable, cost-effective and high-throughput while still assaying a significant number of SNP loci. A range of crop species with differing levels of ploidy and degree of inbreeding/outbreeding were chosen, including perennial ryegrass, a diploid outbreeding forage grass; phalaris, a putative segmental allotetraploid outbreeding forage grass; lentil, a diploid inbreeding grain legume; and canola, an allotetraploid partially outbreeding oilseed. GBS-t was validated as a simple and largely automated, cost-effective method which generates sufficient SNPs (from 89 738 to 231 977) with acceptable levels of missing data and even genome coverage from c. 3 million sequence reads per sample. GBS-t is therefore a broadly applicable system suitable for many crops, offering advantages over other systems. The correct choice of subsequent sequence analysis software is important, and the bioinformatics process should be iterative and tailored to the specific challenges posed by ploidy variation and extent of heterozygosity.

Overexpression of genes associated with hypoxia in cattle adapted to Trans Himalayan region of Ladakh.

Ladakh is an important part of the Trans-Himalayan region located between the Kunlun mountain range in the north and the main Great Himalayas to the south in the state of Jammu and Kashmir of India. The local cattle from Leh and Ladakh region, known as "Ladakhi cattle" is a unique germplasm having an excellent adaptation potential to high altitude hypobaric stress. In the present study, an effort was made to evaluate the transcriptional pattern of hypoxia inducing factor-1 (HIF-1) and several of its regulated genes in PBMCs of local Ladakhi cattle, Holstein Frisian crosses, Jersey (exotic) maintained at high altitude region and Sahiwal (Bos indicus) and Karan Fries (cross bred) cattle maintained in tropical environment. The combined data set indicated increased expression of HIF-1 and its regulated genes viz., glucose transporter 1 (GLUT1), vascular endothelial growth factor (VEGF), and hexokinase (HK2) in high altitude cattle indicating their importance in maintaining cellular homeostasis during high altitude hypoxia. The data indicated that hypoxia associated genes accumulated under hypoxic conditions are part of an essential adaptive component for adaptation to the high altitude of the trans-Himalayan region. In contrary, higher expression of molecular chaperons' viz., HSP70 and HSP90 in tropically adapted cattle give tolerance to high ambient temperature prevalent in tropical condition. In conclusion, HIF-1 and its regulatory genes could be termed as important candidates for producing homeostatic responses to hypoxia in cattle populations reared in higher altitudes of the Trans-Himalayan region.

Devising a method towards development of early warning tool for detection of malaria outbreak.

Background & objectives: Uncertainty often arises in differentiating seasonal variation from outbreaks of malaria. The present study was aimed to generalize the theoretical structure of sine curve for detecting an outbreak so that a tool for early warning of malaria may be developed. Methods: A 'case/mean-ratio scale' system was devised for labelling the outbreak in respect of two diverse districts of Assam and Rajasthan. A curve-based method of analysis was developed for determining outbreak and using the properties of sine curve. It could be used as an early warning tool for Plasmodium falciparum malaria outbreaks. Result: In the present method of analysis, the critical Cmax(peak value of sine curve) value of seasonally adjusted curve for P. falciparum malaria outbreak was 2.3 for Karbi Anglong and 2.2 for Jaisalmer districts. On case/mean-ratio scale, the Cmax value of malaria curve between Cmaxand 3.5, the outbreak could be labelled as minor while >3.5 may be labelled as major. In epidemic years, with mean of case/mean ratio of ≥1.00 and root mean square (RMS) ≥1.504 of case/mean ratio, outbreaks can be predicted 1-2 months in advance. Interpretation & conclusions: The present study showed that in P. falciparum cases in Karbi Anglong (Assam) and Jaisalmer (Rajasthan) districts, the rise in Cmaxvalue of curve was always followed by rise in average/RMS or both and hence could be used as an early warning tool. The present method provides better detection of outbreaks than the conventional method of mean plus two standard deviation (mean+2 SD). The identified tools are simple and may be adopted for preparedness of malaria outbreaks.

A study of normal foot abduction across various age groups in children.

INTRODUCTION: The relapse rate after Ponseti method of correction has reduced in recent years which is attributable to the better appreciation of the need to achieve the correct degree of abduction. Ponseti recommended clinical 'overcorrection' of the foot to 700 of abduction. However, no scientific basis for this figure was found in literature. As the indications of Ponseti method extend to older children, we conducted a study to find out the amount of foot abduction to be achieved before applying a foot abduction brace in various age groups. METHODS: The normal 197 feet of children up to eight years of age were considered for study. The measurements included foot-bimalleolar angle in neutral and maximum possible abduction, thigh foot angle and leg foot angle. RESULTS AND DISCUSSION: The foot bimalleolar angle in neutral had a mean of 82.6 degrees and in maximum abduction a mean of 99.0 degrees. The mean leg foot angle was 66.4 degrees and the mean thigh foot angle was 60.5 degrees. It was found that these variables do not change with age. From the study we concluded that achieved abduction should be about 60-70 degrees before applying foot abduction brace in all children till age eight years with clubfeet treated with Ponseti technique, keeping the leg foot angle or the thigh foot angle as a guideline. This is against the common perception of keeping the abduction at 70 degrees for infants and reducing the abduction to 30 to 40 degrees for older children. Both the leg foot angle and thigh foot angles are reliable indicators of correction.

Single Intravenous Dose of Novel Flurbiprofen-Loaded Proniosome Formulations Provides Prolonged Systemic Exposure and Anti-inflammatory Effect.

Vesicular and colloidal delivery systems can be designed to control drug release spatially and temporally to improve drug efficacy and side effect profiles. Niosomes (vesicles prepared from nonionic surfactants in aqueous media) are gaining interest as an alternative vesicular delivery system as they offer advantages such as biocompatibility, chemical stability, low cost, high purity, and versatility. However, the physical stability of niosomes, like other vesicular systems, is limited by vesicle fusion, aggregation, and leakage. Proniosomes (dehydrated powder or gel formulations that spontaneously form niosomes on hydration with aqueous media) can overcome these physical stability problems and are more convenient for sterilization, storage, transport, distribution, and dosing. Proniosomes have mostly been explored for their potential to enhance transdermal and oral absorption. In this study we assess, for the first time, the potential for hydrated proniosomes to sustain systemic exposure and therapeutic effect after intravenous delivery. Proniosomes carrying the anti-inflammatory drug, flurbiprofen, were prepared by spraying different nonionic surfactants (span 20, span 40, and span 60 in varying ratios with span 80) and cholesterol onto a sorbitol carrier. The proniosome powders were characterized for surface morphology and flow properties. Niosome formation was assessed at three different hydration temperatures (25, 37, and 45 °C), and the niosomes were assessed for vesicle size, entrapment efficiency, and sterility. OLP proniosomes prepared with a high ratio of span 80 to span 20 were found to spontaneously form vesicles of small size and high drug loading on hydration with aqueous media. The OLP derived niosomes successfully sustained in vitro drug release, in vivo pharmacokinetics, and the anti-inflammatory effect of flurbiprofen in an acute (rat paw edema) model of inflammation when compared to a control solution formulation. The study demonstrates that hydrated proniosomes can prolong systemic drug exposure over 3 days and provide a sustained therapeutic effect. The developed proniosomes represent a novel approach to treat acute pain and inflammation with the potential to be administered as a single intravenous dose by a clinician at the time of injury or surgery that provides adequate relief for several days and reduces fluctuations in therapy. Similar systems loaded with different drugs have potential for broader application in anesthesia, anti-infective, antiemetic, and cancer therapy.

Generation and Characterisation of a Reference Transcriptome for Lentil (Lens culinaris Medik.).

RNA-Seq using second-generation sequencing technologies permits generation of a reference unigene set for a given species, in the absence of a well-annotated genome sequence, supporting functional genomics studies, gene characterisation and detailed expression analysis for specific morphophysiological or environmental stress response traits. A reference unigene set for lentil has been developed, consisting of 58,986 contigs and scaffolds with an N50 length of 1719 bp. Comparison to gene complements from related species, reference protein databases, previously published lentil transcriptomes and a draft genome sequence validated the current dataset in terms of degree of completeness and utility. A large proportion (98%) of unigenes were expressed in more than one tissue, at varying levels. Candidate genes associated with mechanisms of tolerance to both boron toxicity and time of flowering were identified, which can eventually be used for the development of gene-based markers. This study has provided a comprehensive, assembled and annotated reference gene set for lentil that can be used for multiple applications, permitting identification of genes for pathway-specific expression analysis, genetic modification approaches, development of resources for genotypic analysis, and assistance in the annotation of a future lentil genome sequence.

Insights into phosphoethanolamine cellulose synthesis and secretion across the Gram-negative cell envelope.

Phosphoethanolamine (pEtN) cellulose is a naturally occurring modified cellulose produced by several Enterobacteriaceae. The minimal components of the E. coli cellulose synthase complex include the catalytically active BcsA enzyme, a hexameric semicircle of the periplasmic BcsB protein, and the outer membrane (OM)-integrated BcsC subunit containing periplasmic tetratricopeptide repeats (TPR). Additional subunits include BcsG, a membrane-anchored periplasmic pEtN transferase associated with BcsA, and BcsZ, a periplasmic cellulase of unknown biological function. While cellulose synthesis and translocation by BcsA are well described, little is known about its pEtN modification and translocation across the cell envelope. We show that the N-terminal cytosolic domain of BcsA positions three BcsG copies near the nascent cellulose polymer. Further, the semicircle's terminal BcsB subunit tethers the N-terminus of a single BcsC protein in a trans-envelope secretion system. BcsC's TPR motifs bind a putative cello-oligosaccharide near the entrance to its OM pore. Additionally, we show that only the hydrolytic activity of BcsZ but not the subunit itself is necessary for cellulose secretion, suggesting a secretion mechanism based on enzymatic removal of translocation incompetent cellulose. Lastly, protein engineering introduces cellulose pEtN modification in orthogonal cellulose biosynthetic systems. These findings advance our understanding of pEtN cellulose modification and secretion.

Molecular insights into phosphoethanolamine cellulose formation and secretion.

Phosphoethanolamine (pEtN) cellulose is a naturally occurring modified cellulose produced by several Enterobacteriaceae. The minimal components of the E. coli cellulose synthase complex include the catalytically active BcsA enzyme, an associated periplasmic semicircle of hexameric BcsB, as well as the outer membrane (OM)-integrated BcsC subunit containing periplasmic tetratricopeptide repeats (TPR). Additional subunits include BcsG, a membrane-anchored periplasmic pEtN transferase associated with BcsA, and BcsZ, a conserved periplasmic cellulase of unknown biological function. While events underlying the synthesis and translocation of cellulose by BcsA are well described, little is known about its pEtN modification and translocation across the cell envelope. We show that the N-terminal cytosolic domain of BcsA positions three copies of BcsG near the nascent cellulose polymer. Further, the terminal subunit of the BcsB semicircle tethers the N-terminus of a single BcsC protein to establish a trans-envelope secretion system. BcsC's TPR motifs bind a putative cello-oligosaccharide near the entrance to its OM pore. Additionally, we show that only the hydrolytic activity of BcsZ but not the subunit itself is necessary for cellulose secretion, suggesting a secretion mechanism based on enzymatic removal of mislocalized cellulose. Lastly, we introduce pEtN modification of cellulose in orthogonal cellulose biosynthetic systems by protein engineering.

Fenugreek derived diosgenin as an emerging source for diabetic therapy.

Diabetes is a chronic metabolic disease that endangers the entire body's tissues and organs. Diabetes impairs glucose and insulin regulation in the human body by causing pancreatic cell damage. Diabetes modifies pathways such as serine/threonine protein kinase (Akt) and Protein kinase C (PKC)/- glucose transporter 4 (GLUT4), peroxisome proliferator-activated receptor (PPAR) glucose absorption, and inhibits α-amylase and α-glucosidase, Sodium/glucose cotransporter 1 (SGLT-1), and Na+-K+-ATPase activity. Diabetes may also be caused by a decrease in the expression of sterol regulatory element binding protein 1 (SREBP-1) and its target genes, fatty acid synthase (FAS), stearoyl-CoA desaturase-1 (SCD-1), and acetyl-CoA carboxylase α (ACC), as well as a decrease in the levels of C/EBP homologous protein (CHOP), Caspase12, and Caspase3 proteins. Diabetes has long been linked to diseases of the cardiovascular, nervous, skeletal, reproductive, hepatic, ocular, and renal systems. Diosgenin, a steroidal compound derived from fenugreek, aids in the prevention of diabetes by altering cellular pathways in favor of healthy bodily functions. Diosgenin is a new nutraceutical on the market that claims to cure diabetes in particular. This article focuses on diosgenin extraction and purification, fenugreek bioactive compounds, pharmacological properties of diosgenin, mode of action of diosgenin to cure diabetes, and dosages.

Characterization of AICAR transformylase/IMP cyclohydrolase (ATIC) bifunctional enzyme from Candidatus Liberibacer asiaticus.

The bifunctional enzyme, 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) transformylase/inosine monophosphate (IMP) cyclohydrolase (ATIC) is involved in catalyzing penultimate and final steps of purine de novo biosynthetic pathway crucial for the survival of organisms. The present study reports the characterization of ATIC from Candidatus Liberibacer asiaticus (CLasATIC) along with the identification of potential inhibitor molecules and evaluation of cell proliferative activity. CLasATIC showed both the AICAR Transformylase (AICAR TFase) activity for substrates, 10-f-THF (Km, 146.6 µM and Vmax, 0.95 µmol/min/mg) and AICAR (Km, 34.81 µM and Vmax, 0.56 µmol/min/mg) and IMP cyclohydrolase (IMPCHase) activitiy (Km, 1.81 µM and Vmax, 2.87 µmol/min/mg). The optimum pH and temperature were also identified for the enzyme activity. In-silico study has been conducted to identify potential inhibitor molecules through virtual screening and MD simulations. Out of many compounds, HNBSA, diosbulbin A and lepidine D emerged as lead compounds, exhibiting higher binding energy and stability for CLasATIC than AICAR. ITC study reports higher binding affinities for HNBSA and diosbulbin A (Kd, 12.3 µM and 34.2 µM, respectively) compared to AICAR (Kd, 83.4 µM). Likewise, DSC studies showed enhanced thermal stability for CLasATIC in the presence of inhibitors. CD and Fluorescence studies revealed significant conformational changes in CLasATIC upon binding of the inhibitors. CLasATIC demonstrated potent cell proliferative, wound healing and ROS scavenging properties evaluated by cell-based bioassays using CHO cells. This study highlights CLasATIC as a promising drug target with potential inhibitors for managing CLas and its unique cell protective, wound-healing properties for future biotechnological applications.

Insights into substrate coordination and glycosyl transfer of poplar cellulose synthase-8.

Cellulose is an abundant cell wall component of land plants. It is synthesized from UDP-activated glucose molecules by cellulose synthase, a membrane-integrated processive glycosyltransferase. Cellulose synthase couples the elongation of the cellulose polymer with its translocation across the plasma membrane. Here, we present substrate and product-bound cryogenic electron microscopy structures of the homotrimeric cellulose synthase isoform-8 (CesA8) from hybrid aspen (poplar). UDP-glucose binds to a conserved catalytic pocket adjacent to the entrance to a transmembrane channel. The substrate's glucosyl unit is coordinated by conserved residues of the glycosyltransferase domain and amphipathic interface helices. Site-directed mutagenesis of a conserved gating loop capping the active site reveals its critical function for catalytic activity. Molecular dynamics simulations reveal prolonged interactions of the gating loop with the substrate molecule, particularly across its central conserved region. These transient interactions likely facilitate the proper positioning of the substrate molecule for glycosyl transfer and cellulose translocation. Highlights: Cryo-EM structures of substrate and product bound poplar cellulose synthase provide insights into substrate selectivitySite directed mutagenesis signifies a critical function of the gating loop for catalysisMolecular dynamics simulations support persistent gating loop - substrate interactionsGating loop helps in positioning the substrate molecule to facilitate cellulose elongationConserved cellulose synthesis substrate binding mechanism across the kingdoms.