Phosphoenolpyruvate carboxykinase in urine exosomes reflect impairment in renal gluconeogenesis in early insulin resistance and diabetes.

Impaired insulin-induced suppression of renal gluconeogenesis could be a risk for hyperglycemia. Diabetes is associated with elevated renal gluconeogenesis; however, its regulation in early insulin resistance is unclear in humans. A noninvasive marker of renal gluconeogenesis would be helpful. Here, we show that human urine exosomes (uE) contain three gluconeogenic enzymes: phosphoenolpyruvate carboxykinase (PEPCK), fructose 1,6-bisphosphatase, and glucose 6-phosphatase. Their protein levels were positively associated with whole body insulin sensitivity. PEPCK protein in uE exhibited a meal-induced suppression. However, subjects with lower insulin sensitivity had blunted meal-induced suppression. Also, uE from subjects with prediabetes and diabetic rats had higher PEPCK relative to nondiabetic controls. Moreover, uE-PEPCK was higher in drug-naïve subjects with diabetes relative to drug-treated subjects with diabetes. To determine whether increased renal gluconeogenesis is associated with hyperglycemia or PEPCK expression in uE, acidosis was induced in rats by 0.28 M NH4Cl with 0.5% sucrose in drinking water. Control rats were maintained on 0.5% sucrose. At the seventh day posttreatment, gluconeogenic enzyme activity in the kidneys, but not in the liver, was higher in acidotic rats. These rats had elevated PEPCK in their uE and a significant rise in blood glucose relative to controls. The induction of gluconeogenesis in human proximal tubule cells increased PEPCK expression in both human proximal tubules and human proximal tubule-secreted exosomes in the media. Overall, gluconeogenic enzymes are detectable in human uE. Elevated PEPCK and its blunted meal-induced suppression in human urine exosomes are associated with diabetes and early insulin resistance.

Systemic inhibition of miR-451 increases fibrotic signaling and diminishes autophagic response to exacerbate renal damage in Tallyho/Jng mice.

miRNAs provide fine tuning of gene expression via inhibition of translation. miR-451 has a modulatory role in cell cycling via downregulation of mechanistic target of rapamycin. We aimed to test whether chronic systemic inhibition of miR-451 would enhance renal fibrosis (associated with deranged autophagy). Adult TallyHo/Jng mice (obese insulin resistant) were randomized to two treatment groups to receive either miR-451 inhibition [via a locked nucleic acid construct] or a similar scrambled locked nucleic acid control for 8 wk. All mice were fed a high-fat diet (60% kcal from fat) ad libitum and humanely euthanized after 12 wk. Kidneys and blood were collected for analysis. Renal expression of miR-451 was sixfold lower in inhibitor-treated mice compared with control mice. miR-451 inhibition increased kidney weight and collagen and glycogen deposition. Blood chemistry revealed significantly higher Na+ and anion gap (relative metabolic acidosis) in inhibitor-treated mice. Western blot analysis and immunohistochemistry of the kidney revealed that the inhibitor increased markers of renal injury and fibrosis, e.g., kidney injury molecule 1, neutrophil gelatinase-associated lipocalin, transforming growth factor-ß, 14-3-3 protein-ζ, mechanistic target of rapamycin, AMP-activated protein kinase-α, calcium-binding protein 39, matrix metallopeptidase-9, and the autophagy receptor sequestosome 1. In contrast, the inhibitor reduced the epithelial cell integrity marker collagen type IV and the autophagy markers microtubule-associated protein 1A/1B light chain 3B and beclin-1. Taken together, these results support a protective role for miR-451 in reducing renal fibrosis by enhancing autophagy in obese mice.

The P2X7 ion channel is dispensable for energy and metabolic homeostasis of white and brown adipose tissues.

Several studies suggest a role of extracellular adenine nucleotides in regulating adipose tissue functions via the purinergic signaling network. Metabolic studies in mice with global deletion of the purinergic receptor P2X7 on the C57BL/6 background indicate that this receptor has only a minor role in adipose tissue for diet-induced inflammation or cold-triggered thermogenesis. However, recent data show that a polymorphism (P451L) present in C57BL/6 mice attenuates P2X7 receptor function, whereas BALB/c mice express the fully functional P451 allele. To determine the potential role of P2rx7 under metabolic and thermogenic stress conditions, we performed comparative studies using male P2rx7 knockout (KO) and respective wild-type controls on both BALB/c and C57BL/6 backgrounds. Our data show that adipose P2rx7 mRNA levels are increased in obese mice. Moreover, P2rx7 deficiency results in reduced levels of circulating CCL2 and IL6 with a moderate effect on gene expression of pro-inflammatory markers in white adipose tissue and liver of BALB/c and C57BL/6 mice. However, P2X7 expression does not alter body weight, insulin resistance, and hyperglycemia associated with high-fat diet feeding on both genetic backgrounds. Furthermore, deficiency of P2rx7 is dispensable for energy expenditure at thermoneutral and acute cold exposure conditions. In summary, these data show that-apart from a moderate effect on inflammatory cytokines-P2X7 plays only a minor role in inflammatory and thermogenic effects of white and brown adipose tissue even on the BALB/c background.

The Synergy between Palmitate and TNF-α for CCL2 Production Is Dependent on the TRIF/IRF3 Pathway: Implications for Metabolic Inflammation.

The chemokine CCL2 (also known as MCP-1) is a key regulator of monocyte infiltration into adipose tissue, which plays a central role in the pathophysiology of obesity-associated inflammation and insulin resistance. It remains unclear how CCL2 production is upregulated in obese humans and rodents. Because elevated levels of the free fatty acid (FFA) palmitate and TNF-α have been reported in obesity, we studied whether these agents interact to trigger CCL2 production. Our data show that treatment of THP-1 and primary human monocytic cells with palmitate and TNF-α led to a marked increase in CCL2 production compared with either treatment alone. Mechanistically, we found that cooperative production of CCL2 by palmitate and TNF-α did not require MyD88, but it was attenuated by blocking TLR4 or TRIF. IRF3-deficient cells did not show synergistic CCL2 production in response to palmitate/TNF-α. Moreover, IRF3 activation by polyinosinic-polycytidylic acid augmented TNF-α-induced CCL2 secretion. Interestingly, elevated NF-κB/AP-1 activity resulting from palmitate/TNF-α costimulation was attenuated by TRIF/IRF3 inhibition. Diet-induced C57BL/6 obese mice with high FFAs levels showed a strong correlation between TNF-α and CCL2 in plasma and adipose tissue and, as expected, also showed increased adipose tissue macrophage accumulation compared with lean mice. Similar results were observed in the adipose tissue samples from obese humans. Overall, our findings support a model in which elevated FFAs in obesity create a milieu for TNF-α to trigger CCL2 production via the TLR4/TRIF/IRF3 signaling cascade, representing a potential contribution of FFAs to metabolic inflammation.

UCP1 deficiency causes brown fat respiratory chain depletion and sensitizes mitochondria to calcium overload-induced dysfunction.

Brown adipose tissue (BAT) mitochondria exhibit high oxidative capacity and abundant expression of both electron transport chain components and uncoupling protein 1 (UCP1). UCP1 dissipates the mitochondrial proton motive force (Δp) generated by the respiratory chain and increases thermogenesis. Here we find that in mice genetically lacking UCP1, cold-induced activation of metabolism triggers innate immune signaling and markers of cell death in BAT. Moreover, global proteomic analysis reveals that this cascade induced by UCP1 deletion is associated with a dramatic reduction in electron transport chain abundance. UCP1-deficient BAT mitochondria exhibit reduced mitochondrial calcium buffering capacity and are highly sensitive to mitochondrial permeability transition induced by reactive oxygen species (ROS) and calcium overload. This dysfunction depends on ROS production by reverse electron transport through mitochondrial complex I, and can be rescued by inhibition of electron transfer through complex I or pharmacologic depletion of ROS levels. Our findings indicate that the interscapular BAT of Ucp1 knockout mice exhibits mitochondrial disruptions that extend well beyond the deletion of UCP1 itself. This finding should be carefully considered when using this mouse model to examine the role of UCP1 in physiology.

Deletion of insulin receptor in the proximal tubule and fasting augment albumin excretion.

The contribution of proximal tubules (PT) to albumin uptake is now well recognized, however, its regulation is understudied area. There are reports suggesting that insulin resistance is associated with the development of albuminuria in nondiabetic individuals. We have previously reported reduced insulin receptor (IR) expression in renal-tubular-epithelial cells, including PT in various models of insulin resistance. However, the effect of a physiological fall in insulin levels and the role for IR in PT in tubular albumin uptake is not clear. To address these gaps in our understanding, we estimated urine excretion and renal uptake of albumin in fasted and fed C57Bl/6 mice injected with fluorescein isothiocyanate (FITC)-albumin (5 µg/mL/kg body weight, intraperitoneal, n = 6 per group). In addition, we compared spot urine analysis from 33 clinically healthy humans after overnight fasting (when insulin levels are lower than in the fed state) and then at 2 hours after 75 g oral glucose challenge (postprandial). Fasted mice had attenuated renal uptake of FITC-albumin and higher excretion in urine, relative to fed mice ( P = 0.04). Moreover, a significant drop in urine albumin-to-creatinine ratio (ACR) and urine albumin concentration (UAC) was observed in the postprandial state in these subjects ( P = 0.001 and P = 0.017, for ACR and UAC, respectively). The drop was negatively associated with postprandial blood glucose levels (ρ = -0.36, P = 0.03 for ΔUAC and ρ = -0.34, P = 0.05 for ΔACR). To test the role of IR in PT, we analyzed 24-hour urine albumin excretion in male mice with targeted deletion of IR from PT (insulin receptor knockout [IRKO]) and their wild-type (WT) littermates ( n = 7 per group). IRKO mice had significantly higher 24-hour urine albumin excretion relative to WT. Moreover, kidneys from KO mice revealed reduced expression of megalin and cubulin proteins in the PT relative to the WT. We also demonstrated insulin (100 nM) induced albumin internalization in human proximal tubule cells (hPT) and this effect of insulin was attenuated in hydroxy-2-naphthalenylmethylphosphonic acid (100 µM), a tyrosine kinase inhibitor, pretreated hPT. Our findings revealed albumin excretion was attenuated by glucose administration to fasting individuals implying a regulatory role for insulin in PT albumin reabsorption. Thus albuminuria associated with insulin resistance/diabetes may relate not only to glomerular dysfunction but also to impairment in insulin-mediated reabsorption.

Bilateral breast metastasis from pancreatic neuroendocrine tumor: A diagnostic challenge.

Metastatic neuroendocrine tumors to breast are very rare with less than 30 cases reported worldwide. The first case was reported in 1957 according to literature. Among these 13 cases reported as breast lump as initial presentation. As the tumor is not so common, the radiological features are not well defined leading to misdiagnosis. Bilateral breast metastasis from pancreatic neuroendocrine tumors has not been mentioned in the literature. Here, we present a case of pancreatic neuroendocrine tumor with metastasis to bilateral breast.

Thoracoscopic oesophagectomy for end-stage achalasia.

Achalasia cardia is an oesophageal motility disorder characterised by aperistalsis and failure of relaxation of the lower oesophageal sphincter. The management is predominantly palliative with focus on addressing the sphincter that involves either pneumatic dilatation or Heller myotomy which relieves dysphagia in the majority of the cases. End-stage achalasia (ESA) is characterised by failed myotomy, massively dilated and tortuous oesophagus with nutritional deterioration due to progressive dysphagia and vomiting. In these subgroups of patients, oesophagectomy may be the last resort. While oesophagectomy has been described for ESA before, thoracoscopic oesophagectomy has not been reported previously. Hereby, we report our experience of performing minimally invasive oesophagectomy (thoracoscopic) with the gastric pull-up.

Genome-wide analysis of miRNAs and Tasi-RNAs in Zea mays in response to phosphate deficiency.

Globally important cereal crop maize provides important nutritions and starch in dietary foods. Low phosphate (LPi) availability in the soil frequently limits the maize quality and yield across the world. Small non-coding RNAs (Snc-RNAs) play crucial roles in growth and adaptation of plants to the environment. Snc-RNAs like microRNAs (miRs) and trans-acting small interfering RNAs (Tasi-Rs) play important functions in posttranscriptional regulation of gene expression, which controls plant development, reproduction, and biotic/abiotic stress responses. In order to identify the miR and Tasi-R alterations in leaf and root of maize in response to sufficient phosphate and LPi at 3LS and 4LS, the snc-RNA population libraries for 0th, 1st, 2nd, 4th, and 8th day were constructed. These libraries were used for genome-wide alignment and RNA-fold analysis for possible prediction of potential miRs and Tasi-Rs. This study reported 174 known and conserved differentially expressed miRs of 27 miR families of maize plant. In addition, leaf and root specific potential novel miRs representing 155 new families were also discovered. Differentially expressed conserved as well as novel miR functions in root and leaf during early stage of Pi starvation were extensively discussed. Leaf and root specific miRs as well as common miRs with their target genes, participating in different biological, cellular, and metabolic processes were explored. Further, four miR390-directed Tasi-Rs which belong to TAS3 gene family along with other orthologs of Tasi-Rs were also identified. Finally, the study provides an insight into the composite regulatory mechanism of miRs in maize in response to Pi deficiency.

Comparative Evaluation of Tensile Strength in Die Stone Incorporated with Sodium and Calcium Hypochlorite as Disinfectants: An in vitro Study.

AIM: The aim of this study was to evaluate the tensile strength of die stone incorporated with sodium and calcium hypochlorite as disinfectants. MATERIALS AND METHODS: Two commercially available type IV die stone (Kalrock: Kalabhai Karson Pvt., Ltd and Pearlstone: Asian Chemicals) and two commercially available disinfectant solutions (sodium hypochlorite and calcium hypochlorite: Beachem Laboratory Chemical Private Limited, Chennai and Leo Chem Private Limited, Bengaluru) were used in this study, and the tensile strength was measured using Lloyd's Universal Testing Machine. RESULTS: The results show that incorporating the disinfecting solutions decreases the tensile strength of both products. The effect of decreasing tensile strength on type IV gypsum product is seen more in calcium hypochlorite when compared with sodium hypochlorite disinfecting solution, and the tensile strength of Kalrock specimens is higher than Pearlstone specimens after disinfecting with sodium hypochlorite and calcium hypochlorite solution. The statistical results also show significant results in all the groups when compared with the control group. CONCLUSION: The incorporation of sodium and calcium hypochlorite disinfecting solutions is not an encouraging method for both die materials as it reduces the tensile strength of type IV gypsum product. Tensile strength of Kalstone® die material is superior than Pearlstone® die material after mixing with sodium hypochlorite and calcium hypochlorite. CLINICAL SIGNIFICANCE: According to the recommendations of Americans with Disability Act (ADA) and the Centers for Disease Control and Prevention, disinfecting the whole cast without or minimal changes in physical and mechanical properties was the motto of the study. The tensile strength in type IV gypsum product plays a most important role in retrieval of cast from impression, especially in narrow tooth preparation. This study reveals that incorporating method of disinfecting solutions is not recommended as it reduces the tensile strength.

Functional cardiac lipolysis in mice critically depends on comparative gene identification-58.

Efficient catabolism of cellular triacylglycerol (TG) stores requires the TG hydrolytic activity of adipose triglyceride lipase (ATGL). The presence of comparative gene identification-58 (CGI-58) strongly increased ATGL-mediated TG catabolism in cell culture experiments. Mutations in the genes coding for ATGL or CGI-58 in humans cause neutral lipid storage disease characterized by TG accumulation in multiple tissues. ATGL gene mutations cause a severe phenotype especially in cardiac muscle leading to cardiomyopathy that can be lethal. In contrast, CGI-58 gene mutations provoke severe ichthyosis and hepatosteatosis in humans and mice, whereas the role of CGI-58 in muscle energy metabolism is less understood. Here we show that mice lacking CGI-58 exclusively in muscle (CGI-58KOM) developed severe cardiac steatosis and cardiomyopathy linked to impaired TG catabolism and mitochondrial fatty acid oxidation. The marked increase in ATGL protein levels in cardiac muscle of CGI-58KOM mice was unable to compensate the lack of CGI-58. The addition of recombinant CGI-58 to cardiac lysates of CGI-58KOM mice completely reconstituted TG hydrolytic activities. In skeletal muscle, the lack of CGI-58 similarly provoked TG accumulation. The addition of recombinant CGI-58 increased TG hydrolytic activities in control and CGI-58KOM tissue lysates, elucidating the limiting role of CGI-58 in skeletal muscle TG catabolism. Finally, muscle CGI-58 deficiency affected whole body energy homeostasis, which is caused by impaired muscle TG catabolism and increased cardiac glucose uptake. In summary, this study demonstrates that functional muscle lipolysis depends on both CGI-58 and ATGL.

To check the antibacterial efficacy against Enterococcus faecalis by various ayurvedic oils used as a solvent in endodontic sealers - An in vitro study.

Aim: The aim of the study is to check the antibacterial efficacy of various ayurvedic oils used as a solvent with zinc oxide for preparing endodontic sealers. Materials and Methods: Forty-five extracted premolars were taken and were cut coronally and apically such that 7 mm of tooth specimen was prepared. Teeth were sterilized by autoclaving inoculated with Enterococcus faecalis and incubated for 24 h. The specimens were divided into three groups of 15 each. Group 1 - ZnO powder + Eugenol, Group 2 - ZnO powder + Aremidadi Oil, and Group 3 - ZnO powder + Dashmool oil. Bacterial growth in each specimen was calculated before and after sealer application and noted as the initial and final colony count. The antimicrobial effect of each sealer was measured by calculating the percentage reduction in colony count (%). One-way analysis of variance and post hoc tests will be used for statistical analysis. Results: The Zn + Arimedadi oil group showed the maximum antibacterial effect among the sealers tested and the Zn + eugenol sealer showed the least antimicrobial effect In comparison, there was a statistically significant difference between all the groups. Conclusion: Ayurvedic oil-based root canal sealers showed better antibacterial efficacy than eugenol-based sealers. Arimedadi oil showed the highest antibacterial activity against E. faecalis and Eugenol showed the least when used as a solvent.

Long-term application of organic manures and chemical fertilizers improve the organic carbon and microbiological properties of soil under pearl millet-wheat cropping system in North-Western India.

An on-going long term field experiment started in Rabi 1995 at the Research Farm of the Department of Soil Science, Chaudhary Charan Singh Haryana Agricultural University, Hisar, Haryana (India) under the pearl millet-wheat cropping system was selected to study the effect of long-term application of organic manures and fertilizers on soil organic carbon and microbiological properties. Highest soil organic carbon (SOC: 1.18 %), dissolved OC (DOC: 64.74 mg kg-1) content, microbial biomass C (MBC: 618.40 mg kg-1), dehydrogenase (DHA: 72.83 µg TPF g-1 24 hr-1), alkaline phosphatase (APA: 685.44 µg PNP g-1 soil hr-1) and aryl sulfatase (ASA: 12.56 µg PNP g-1 soil hr-1) activities were observed with the application of 15 Mg FYM+150 kg N+30 kg P2O5 ha-1. Integrated application of chemical fertilizers with pressmud showed superiority in the improvement of microbial biomass nitrogen (MBN: 73.73 mg kg-1) and urease activity (69.54 µg NH4+ g-1 hr-1) than FYM or poultry manure plus NP. Beneficial impacts of the sole application of organic manures on SOC, DOC, MBC content, DHA, APA, and ASA were found in order as: FYM > pressmud > poultry manure. Impacts of nutrient management practices on soil carbon fractions decreased with depth. Poultry manure application, either alone or in combination with NP fertilizers was inferior to FYM and pressmud. The SOC had a positive relationship with MBC (R2 = 0.95) and MBN (R2 = 0.75) and, also showed a highly positive and significant correlation with microbiological properties of soil. This dynamic equilibrium among soil properties indicated that the nutrient management practices that improve SOC could lead to improve soil fertility and accrued microbiological properties in these soils. This study revealed that conjuctive use of organic manures and chemical fertilizers have positive impact on soil fertility and microbiological properties as compared to sole application of organic manures or fertilizers; and among organic manures, FYM was superior to pressmud followed by poultry manure.

Cardiac-specific overexpression of perilipin 5 provokes severe cardiac steatosis via the formation of a lipolytic barrier.

Cardiac triacylglycerol (TG) catabolism critically depends on the TG hydrolytic activity of adipose triglyceride lipase (ATGL). Perilipin 5 (Plin5) is expressed in cardiac muscle (CM) and has been shown to interact with ATGL and its coactivator comparative gene identification-58 (CGI-58). Furthermore, ectopic Plin5 expression increases cellular TG content and Plin5-deficient mice exhibit reduced cardiac TG levels. In this study we show that mice with cardiac muscle-specific overexpression of perilipin 5 (CM-Plin5) massively accumulate TG in CM, which is accompanied by moderately reduced fatty acid (FA) oxidizing gene expression levels. Cardiac lipid droplet (LD) preparations from CM of CM-Plin5 mice showed reduced ATGL- and hormone-sensitive lipase-mediated TG mobilization implying that Plin5 overexpression restricts cardiac lipolysis via the formation of a lipolytic barrier. To test this hypothesis, we analyzed TG hydrolytic activities in preparations of Plin5-, ATGL-, and CGI-58-transfected cells. In vitro ATGL-mediated TG hydrolysis of an artificial micellar TG substrate was not inhibited by the presence of Plin5, whereas Plin5-coated LDs were resistant toward ATGL-mediated TG catabolism. These findings strongly suggest that Plin5 functions as a lipolytic barrier to protect the cardiac TG pool from uncontrolled TG mobilization and the excessive release of free FAs.

Studies on the substrate and stereo/regioselectivity of adipose triglyceride lipase, hormone-sensitive lipase, and diacylglycerol-O-acyltransferases.

Adipose triglyceride lipase (ATGL) is rate-limiting for the initial step of triacylglycerol (TAG) hydrolysis, generating diacylglycerol (DAG) and fatty acids. DAG exists in three stereochemical isoforms. Here we show that ATGL exhibits a strong preference for the hydrolysis of long-chain fatty acid esters at the sn-2 position of the glycerol backbone. The selectivity of ATGL broadens to the sn-1 position upon stimulation of the enzyme by its co-activator CGI-58. sn-1,3 DAG is the preferred substrate for the consecutive hydrolysis by hormone-sensitive lipase. Interestingly, diacylglycerol-O-acyltransferase 2, present at the endoplasmic reticulum and on lipid droplets, preferentially esterifies sn-1,3 DAG. This suggests that ATGL and diacylglycerol-O-acyltransferase 2 act coordinately in the hydrolysis/re-esterification cycle of TAGs on lipid droplets. Because ATGL preferentially generates sn-1,3 and sn-2,3, it suggests that TAG-derived DAG cannot directly enter phospholipid synthesis or activate protein kinase C without prior isomerization.

Comparative evaluation of canal transportation and centering ability of rotary and reciprocating file systems using cone-beam computed tomography: An in vitro study.

Aim: The aim of the study was to evaluate the canal transportation and centering ability of rotary and reciprocating file systems using cone-beam computed tomography. Materials and Methods: Mesiobuccal canals of sixty mandibular molars were selected for the study. Canals of length 19 mm, curvature of 10°-12°, and uncalcified with fully formed apex were selected. Canals were randomly divided into three groups of 20 teeth, and canal preparation with the WaveOne Gold, TruNatomy, and One Curve systems was performed according to the manufacturers' instructions. Cone-beam computed tomographic images were taken before and after instrumentation in the same position for comparative analysis. Statistical Analysis Used: Apical transportation was calculated at the distances of 2, 3, and 4 mm from the apex. Tukey's post hoc test and unpaired "t"-tests were used to statistically analyze the data. Results: WaveOne Gold caused less canal transportation and better-centering ability than TruNatomy and One Curve at all the three levels; there was a significant difference in canal transportation and centering ability among all the groups as well as all the three levels, i.e., 2, 3, and 4 mm from the apex. Conclusion: WaveOne Gold (Reciprocating) reported less canal transportation and better-centering ability than rotary instruments TruNatomy and One Curve (Rotary) at all the three levels.

Enhanced degradation of Congo-red dye by Cr3+ doped α-Fe2O3 nano-particles under sunlight and industrial wastewater treatment.

Considering the increasing amount of water pollution, nanocomposite advances for the effective elimination of hazardous pollutants are still needed. α-Fe2O3, Cr0·5Fe1·5O3 and CrFeO3 nanoparticles were synthesized via an eco-friendly material synthesis i. e hydrothermal route without using any precipitating agent and were studied to remove congo-red dye using photocatalytic properties. X-ray diffraction (XRD), Transmission electron microscopy (TEM), Field emission scanning electron microscopy (FESEM), Fourier transform infrared spectroscopy (FESEM), Brunauer-Emmett-Teller (BET), X-ray photoelectron spectroscopy (XPS) characterizations have been performed to know about the material structure and properties of synthesized samples. High efficiency (95.2%) of degradation was achieved under sunlight using a very low amount of CrFeO3 catalyst (0.2 g/L) at a 6-pH level of dye and was confirmed using UV spectroscopy, TOC (84%), LC-HRMS. Also, the potential to degrade the CR dye was concluded from the high rate of BOD5/COD. The results showed a significant enhancement in the degradation of α-Fe2O3 from 52.3% to 95.2% in a short duration of 15 min by introducing chromium as a dopant. The doping of chromium influenced the major factors responsible for the photocatalytic activity such as the increase in range of absorbance, increased e--h+ pair separation, improvement in the charge transfer process and active site formation which significantly enhanced the process of degradation. We found that the Cr-doped α-Fe2O3 nanomaterial could effectively remove dyes, such as congo-red, from industrial water-waste.

Rare fatal effect of combined use of sildenafil and alcohol leading to Cerebrovascular Accident.

Sildenafil is the first internationally approved drug for erectile dysfunction. Unsupervised and non-prescribed use of sildenafil among young Indian population has increased in last few years. Sildenafil helps in erection of penis by inhibiting the action of Phosphodiesterase-5 (PDE-5) enzyme, present in the vasculature of corpus cavernosum muscle and lengthens the duration of erection. Documented adverse effects of sildenafil are headache, flushing, nasal congestion, dyspepsia, and slight decrease in systolic and diastolic blood pressure. We present a rare case of sudden death due to cerebrovascular hemorrhage after sildenafil use and concomitant alcohol intake. The history is that a 41-year-old male with no significant past medical and surgical history was staying at a hotel room with a female friend; he had consumed 2 tablets of sildenafil (50 mg each) and alcohol at night. Next morning, he developed uneasiness following which he was taken to the Hospital where he was declared dead on arrival. The important autopsy findings include edematous brain with about 300 g of clotted blood in the right basal ganglia extending to bilateral ventricles, and in pons region. Other significant findings on microscopic examination were hypertrophic ventricular wall of heart, fatty changes in liver and acute tubular necrosis and hypertensive changes in the kidney. The findings are discussed in the light of the literature about the lethal complications of combined use of sildenafil and alcohol including cerebrovascular accidents. As a forensic pathologist it is the duty of the doctor to execute meticulous autopsy along with ancillary investigations including toxicological analysis and to correlate all these findings to determine the possible effects of drugs when present, so as to gather knowledge about potentially fatal drugs and further create public awareness regarding the same.

Unveiling Diversity for Quality Traits in the Indian Landraces of Horsegram [Macrotyloma uniflorum (Lam.) Verdc.].

Horsegram (Macrotyloma uniflorum [Lam.] Verdc.) is an underutilized pulse crop primarily cultivated in South Asian countries like India, Nepal, and Sri Lanka. It offers various nutraceutical properties and demonstrates remarkable resilience to both biotic and abiotic stresses. As a result, it has emerged as a promising crop for ensuring future food and nutritional security. The purpose of this study was to assess the nutritional profile of 139 horsegram germplasm lines obtained from 16 Indian states that were conserved at the National Gene Bank of India. Standard analytical methods, including those provided by the Association of Official Analytical Chemists (AOAC), were used for this investigation. The study revealed substantial variability in essential nutrients, such as protein (ranging from 21.8 to 26.7 g/100 g), starch (ranging from 26.2 to 33.0 g/100 g), total soluble sugars (TSSs) (ranging from 0.86 to 12.1 g/100 g), phenolics (ranging from 3.38 to 11.3 mg gallic acid equivalents (GAEs)/g), and phytic acid content (ranging from 1.07 to 21.2 mg/g). Noteworthy correlations were observed, including a strong positive correlation between sugars and phenols (r = 0.70) and a moderate negative correlation between protein and starch (r = -0.61) among the studied germplasm lines. Principal component analysis (PCA) highlighted that the first three principal components contributed to 88.32% of the total variability, with TSSs, phytates, and phenols emerging as the most significant contributors. The cluster analysis grouped the accessions into five clusters, with cluster III containing the accessions with the most desirable traits. The differential distribution of the accessions from north India into clusters I and III suggested a potential geographical influence on the adaptation and selection of genes. This study identified a panel of promising accessions exhibiting multiple desirable traits. These specific accessions could significantly aid quality breeding programs or be directly released as cultivars if they perform well agronomically.

Conformational mapping and energetics of saccharide-aromatic residue interactions: implications for the discrimination of anomers and epimers and in protein engineering.

Aromatic residues play a key role in saccharide-binding sites. Experimental studies have given an estimate of the energetics of saccharide-aromatic residue interactions. In this study, dependence of the energetics on the mutual position-orientation (PO) of saccharide and aromatic residue has been investigated by geometry optimization of a very large number (164) of complexes at MP2/6-31G(d,p) level of theory. The complexes are of Tyr and Phe analogs with α/ß-D-Glc, ß-D-Gal, α-D-Man and α/ß-L-Fuc. A number of iso-energy POs are found for the complexes of all six saccharides. Stacking and non-stacking modes of binding are found to be of comparable strengths. In general, complexes of p-OHTol are stronger than those of Tol, and those dominated by OH···O interactions are more stable than ones dominated by CH···π interactions. The strengths of OH···O/π interactions, but not those of CH···π, show large variations. Even though an aromatic residue has a large variety of POs to interact with a saccharide, distinct preferences are found due to anomeric and epimeric differences. An aromatic residue can interact from either the a- or b-face of Glc, but only through the b-face with Gal, its C4-epimer. In contrast, stacking interaction with Man (C2-epimer of Glc) requires the participation of the -CH(2)OH group and free rotation of this group, as is observed in solution, precludes all modes of stacking interactions. It is also found that an aromatic residue can be strategically placed either to discriminate or to accommodate (i) anomers of Glc and of Fuc and (ii) Gal/Fuc. Thus, analysis of the optimized geometries of by far the largest number of complexes, and with six different saccharides, at this level of theory has given insights into how Nature cleverly uses aromatic residues to fine tune saccharide specificities of proteins. These are of immense utility for protein engineering and protein design studies.