Further evidence that peritraumatic 17ß-estradiol levels influence chronic posttraumatic pain outcomes in women, data from both humans and animals.

ABSTRACT: Chronic posttraumatic pain (CPTP) is common after traumatic stress exposure (TSE) and disproportionately burdens women. We previously showed across 3 independent longitudinal cohort studies that, in women, increased peritraumatic 17ß-estradiol (E2) levels were associated with substantially lower CPTP over 1 year. Here, we assessed this relationship in a fourth longitudinal cohort and also assessed the relationship between E2 and CPTP at additional time points post-TSE. Furthermore, we used a well-validated animal model of TSE to determine whether exogenous E2 administration protects against mechanical hypersensitivity. Using nested samples and data from the Advancing Understanding of RecOvery afteR traumA study (n = 543 samples, 389 participants), an emergency department-based prospective study of TSE survivors, we assessed the relationship between circulating E2 levels and CPTP in women and men using multivariate repeated-measures mixed modeling. Male and ovariectomized female Sprague Dawley rats were exposed to TSE and administered E2 either immediately after or 3 days post-TSE. Consistent with previous results, we observed an inverse relationship between peritraumatic E2 and longitudinal CPTP in women only (ß = -0.137, P = 0.033). In animals, E2 protected against mechanical hypersensitivity in female ovariectomized rats only if administered immediately post-TSE. In conclusion, peritraumatic E2 levels, but not those at post-TSE time points, predict CPTP in women TSE survivors. Administration of E2 immediately post TSE protects against mechanical hypersensitivity in female rats. Together with previous findings, these data indicate that increased peritraumatic E2 levels in women have protective effects against CPTP development and suggest that immediate post-TSE E2 administration in women could be a promising therapeutic strategy for reducing risk of CPTP.

CRISPR-Cas for genome editing: Classification, mechanism, designing and applications.

Clustered regularly interspersed short pallindromic repeats (CRISPR) and CRISPR associated proteins (Cas) system (CRISPR-Cas) came into light as prokaryotic defence mechanism for adaptive immune response. CRISPR-Cas works by integrating short sequences of the target genome (spacers) into the CRISPR locus. The locus containing spacers interspersed repeats is further expressed into small guide CRISPR RNA (crRNA) which is then deployed by the Cas proteins to evade the target genome. Based on the Cas proteins CRISPR-Cas is classified according to polythetic system of classification. The characteristic of the CRISPR-Cas9 system to target DNA sequences using programmable RNAs has opened new arenas due to which today CRISPR-Cas has evolved as cutting end technique in the field of genome editing. Here, we discuss about the evolution of CRISPR, its classification and various Cas systems including the designing and molecular mechanism of CRISPR-Cas. Applications of CRISPR-Cas as a genome editing tools are also highlighted in the areas such as agriculture, and anticancer therapy. Briefly discuss the role of CRISPR and its Cas systems in the diagnosis of COVID-19 and its possible preventive measures. The challenges in existing CRISP-Cas technologies and their potential solutions are also discussed briefly.

Novel catalytic potential of a hyperthermostable mono­copper oxidase (LPMO-AOAA17) for the oxidation of lignin monomers and depolymerisation of lignin dimer in aqueous media.

Lytic polysaccharide monooxygenase (LPMO) are mono­copper enzymes known for the oxidative cleavage of recalcitrant polysaccharides with their intriguing and unique catalytic chemistry. Such impeccable oxidation potential has made them highly valuable in the enzymatic consortia for the degradation of ligno-cellulosic biomass. Bioinformatic analysis has revealed an unannotated LPMO gene in the genome of A. oryzae. Multiple sequence alignment showed the presence of conserved "histidine brace" of LPMO in the amino acid sequence of the enzyme. The enzyme, named as LPMO-AOAA17 was recombinantly expressed in E. coli BL21 and characterised for its substrate specificity. Recombinant enzyme did not show any characteristic cleavage of polysaccharides. However, it was found to be oxidising broad range of phenolic and non-phenolic monomers of lignin. Biochemical study revealed the optimum activity of LPMO-AOAA17 at pH 7 and was highly stable and active at 100 °C. The enzyme LPMO-AOAA17 was also observed to be stable after autoclaving at 121 °C and 15 psi. Thermal stability of the LPMO-AOAA17 was further confirmed through differential scanning calorimetry. GC-MS analysis has confirmed the catalysis of LPMO-AOAA17 for the depolymerisation of lignin dimer, guaicyl glycerol ß-guaicyl ether into guaiacol. This study has first time documented the identification of a hyperthermostable LPMO for oxidative cleavage of ß-O-4 linkage of lignin compounds to form aromatic products in aqueous media.

Pathogenesis of Celiac Disease and Other Gluten Related Disorders in Wheat and Strategies for Mitigating Them.

Wheat is a major cereal crop providing energy and nutrients to the billions of people around the world. Gluten is a structural protein in wheat, that is necessary for its dough making properties, but it is responsible for imparting certain intolerances among some individuals, which are part of this review. Most important among these intolerances is celiac disease, that is gluten triggered T-cell mediated autoimmune enteropathy and results in villous atrophy, inflammation and damage to intestinal lining in genetically liable individuals containing human leukocyte antigen DQ2/DQ8 molecules on antigen presenting cells. Celiac disease occurs due to presence of celiac disease eliciting epitopes in gluten, particularly highly immunogenic alpha-gliadins. Another gluten related disorder is non-celiac gluten-sensitivity in which innate immune-response occurs in patients along with gastrointestinal and non-gastrointestinal symptoms, that disappear upon removal of gluten from the diet. In wheat allergy, either IgE or non-IgE mediated immune response occurs in individuals after inhalation or ingestion of wheat. Following a life-long gluten-free diet by celiac disease and non-celiac gluten-sensitivity patients is very challenging as none of wheat cultivar or related species stands safe for consumption. Hence, different molecular biology, genetic engineering, breeding, microbial, enzymatic, and chemical strategies have been worked upon to reduce the celiac disease epitopes and the gluten content in wheat. Currently, only 8.4% of total population is affected by wheat-related issues, while rest of population remains safe and should not remove wheat from the diet, based on false media coverage.

A study of risk factors and foot care behavior among diabetics.

BACKGROUND: Diabetic foot results in considerable morbidity and mortality in developing countries and the prevalence of diabetes is expected to increase further in the next decades in these countries. Diabetic ulcers are the most common foot injuries leading to lower extremity amputation. Family physicians have a pivotal role in the prevention or early diagnosis of diabetic foot complications. Patient education regarding foot hygiene, nail care and proper footwear is crucial to reducing the risk of an injury that can lead to ulcer formation. MATERIALS AND METHODS: This is a prospective study carried out from July 2013 to September 2015. Fifty patients of Diabetes with foot ulcer and two hundred without foot ulcers were examined. Risk factors and clinical profile of patients were studied which included age, gender, duration of diabetes, BMI, smoking, random BSLs history, hypertension, glycated haemoglobin levels, lipid profile, history of loss of sensation and history of amputation. MNSI questionnaire and MNSI practical assessment for neuropathy were administered to diabetic patients along with a pre-structured questionnaire regarding foot care practices. RESULTS: In this study significant risk factors were peripheral neuropathy, peripheral vascular disease, gender, loss of sensation, duration of diabetes and smoking. MNSI questionnaire and practical assessment scores were higher in foot ulcer patients. Poor foot care practices were observed in patients with diabetic foot ulcer patients. CONCLUSION: Diabetic foot ulcers were more common in elderly males. Peripheral neuropathy, peripheral vascular disease, Smoking, trauma, duration of diabetes mellitus and high levels of glycated haemoglobin had significant association with occurrence of foot ulcers. MNSI scores had a high predictive value for development of foot ulcers amongst diabetics. Awareness regarding foot care was poor which underlines need to promote practice of foot care amongst diabetic population.

An RNA aptamer with high affinity and broad specificity for zinc finger proteins.

A class of RNA aptamers that demonstrates a high affinity for a large variety of C(2)H(2) zinc finger proteins was isolated from a library of random RNA sequences by the zinc finger protein TFIIIA. These aptamers have one or more copies of the consensus sequence GGGUGGG, which is part of a putative hairpin loop in the proposed structure of the most abundant aptamer, RNA1. Binding of zinc finger proteins to RNA1 relies upon zinc-dependent folding of the protein, the affinity of an individual protein for RNA1 being determined by the number of tandem zinc finger motifs. The properties of RNA1 were compared to the properties of two other aptamers from the same selection experiment: RNA21, which binds to some but not all zinc finger proteins tested, and RNA22, which binds only to the 5S rRNA binding zinc finger proteins TFIIIA and p43. The binding of three different zinc finger proteins to RNA1 was compared, and the results indicate that the RNA1-protein interaction occurs by several distinct mechanisms. Mutagenesis of RNA1 confirmed that the GGGUGGG consensus sequence presented in a hairpin conformation is required for high-affinity binding of zinc finger proteins.

Contribution of individual amino acids to the 5S RNA binding activity of the Xenopus zinc finger protein p43.

Xenopus zinc finger protein p43 binds to 5S RNA in immature oocytes to form a 42S ribonucleoprotein storage particle. To determine the role of individual zinc fingers of the protein in this RNA binding activity, a series of deletion and substitution mutants of p43 were constructed. The effects of the various mutations on the RNA binding activity of p43 were determined using a quantitative equilibrium binding assay. The results indicate that zinc fingers 1 and 4 of p43 are essential for the binding of the protein to 5S RNA. In the case of finger 1, four amino acids key to RNA binding are found on the same face of the alpha-helix, while in the case of finger 4, two key residues are clustered at the start of the alpha-helix. The similarities and differences in the mechanisms by which fingers 1 and 4 of p43 interact with 5S RNA are compared to the interaction of the zinc fingers of Xenopus transcription factor IIIA with 5S RNA.