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Environmental enteric dysfunction (EED), a small intestinal disorder found at a high prevalence in stunted children, is associated with gut mucosal barrier disruption and decreased absorptive capacity due to reduced intact small intestinal villi. To test the hypothesis that intergenerational transmission of a perturbed small intestinal microbiota contributes to undernutrition by inducing EED, we characterized two consortia of bacterial strains cultured from duodenal aspirates from stunted Bangladeshi children with EED, one of which induced local and systemic inflammation in gnotobiotic female mice. Offspring of dams that received this inflammatory consortium exhibited immunologic changes along their gut that phenocopied features of EED in children. Single nucleus plus bulk RNA-sequencing revealed alterations in inter-cellular signaling pathways related to intestinal epithelial cell renewal, barrier integrity and immune function while analyses of cerebral cortex disclosed alterations in glial- and endothelial-neuronal signaling pathways that regulate neural growth/axonal guidance, angiogenesis and inflammation. Analysis of ultrasonic vocalization calls in gnotobiotic P5-P9 pups indicated increased arousal and perturbed neurodevelopment in the offspring of dams harboring the inflammation-inducing consortium. Cohousing experiments and follow-up screening of candidate disease-promoting bacterial isolates identified a strain typically found in the oral microbiota (Campylobacter concisus) as a contributor to enteropathy. Given that fetal growth was also impaired in the dams with the consortium that induced enteropathy, this preclinical model allows the effects of the human small intestinal microbiota on both pre- and postnatal development to be ascertained, setting the stage for identification of small intestinal microbiota-targeted therapeutics for (intergenerational) undernutrition.

Nasal packing is one of the common procedures done following various intranasal surgeries in order to prevent the various complications that can arise post-operatively. Some of the complications noted include bleeding from the nose, septal hematoma, septal abscess and formation of mucosal adhesions. There are different types of nasal packing materials available. Few of the commonly used nasal packing material are ribbon gauzes impregnated with antibiotics, merocel, gel foam, glove fingers and Vaseline gauze. This study compared the effectiveness of Merocel nasal packing with Glove finger nasal packing after Septoplasty and Polypectomy. Merocel, a widely used expandable non-absorbable material, was compared with conventional ribbon gauze packs inserted into glove fingers. Results showed no significant differences in post-operative bleeding, pain, discomfort, mucociliary clearance, or late complications between the two groups. However, a slightly earlier return of mucociliary clearance in patients packed with Glove finger nasal packs was observed. Therefore, it can be concluded that both Merocel and Glove finger nasal packs are equally effective in managing post-operative complications following these procedures., Competing Interests: Conflict of InterestNA., (© Association of Otolaryngologists of India 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Stem cells have lower facultative heterochromatin as defined by trimethylation of histone H3 lysine 27 (H3K27me3) compared to differentiated cells. However, the mechanisms underlying these differential H3K27me3 levels remain elusive. Because H3K27me3 levels are diluted two-fold in every round of replication and then restored through the rest of the cell cycle, we reasoned that the cell cycle length could be a key regulator of total H3K27me3 levels. Here, we propose that a fast cell cycle restricts H3K27me3 levels in stem cells. To test this model, we determined changes to H3K27me3 levels in mESCs globally and at specific loci upon G1 phase lengthening - accomplished by thymidine block or growth in the absence of serum (with the "2i medium"). H3K27me3 levels in mESC increase with G1 arrest when grown in serum and in 2i medium. Additionally, we observed via CUT&RUN and ChIP-seq that regions that gain H3K27me3 in G1 arrest and 2i media overlap, supporting our model of cell cycle length as a critical regulator of the stem cell epigenome and cellular identity. Furthermore, we demonstrate the inverse effect - that G1 shortening in differentiated cells results in a loss of H3K27me3 levels. Finally, in tumor cells with extreme H3K27me3 loss, lengthening of the G1 phase leads to H3K27me3 recovery despite the presence of the dominant negative, sub-stoichiometric H3.1K27M mutation. Our results indicate that G1 length is an essential determinant of H3K27me3 landscapes across diverse cell types.

Acute systemic inflammation critically alters the function of the immune system, often promoting myelopoiesis at the expense of lymphopoiesis. In the thymus, systemic inflammation results in acute thymic atrophy and, consequently, impaired T-lymphopoiesis. The mechanism by which systemic inflammation impacts the thymus beyond suppressing T-cell development is still unclear. Here, we describe how the synergism between TL1A and IL-18 suppresses T-lymphopoiesis to promote thymic myelopoiesis. The protein levels of these two cytokines were elevated in the thymus during viral-induced thymus atrophy infection with murine cytomegalovirus (MCMV) or pneumonia virus of mice (PVM). In vivo administration of TL1A and IL-18 induced acute thymic atrophy, while thymic neutrophils expanded. Fate mapping with Ms4a3-Cre mice demonstrated that thymic neutrophils emerge from thymic granulocyte-monocyte progenitors (GMPs), while Rag1-Cre fate mapping revealed a common developmental path with lymphocytes. These effects could be modeled ex vivo using neonatal thymic organ cultures (NTOCs), where TL1A and IL-18 synergistically enhanced neutrophil production and egress. NOTCH blockade by the LY411575 inhibitor increased the number of neutrophils in the culture, indicating that NOTCH restricted steady-state thymic granulopoiesis. To promote myelopoiesis, TL1A, and IL-18 synergistically increased GM-CSF levels in the NTOC, which was mainly produced by thymic ILC1s. In support, TL1A- and IL-18-induced granulopoiesis was completely prevented in NTOCs derived from Csf2rb -/- mice and by GM-CSFR antibody blockade, revealing that GM-CSF is the essential factor driving thymic granulopoiesis. Taken together, our findings reveal that TL1A and IL-18 synergism induce acute thymus atrophy while  promoting extramedullary thymic granulopoiesis in a NOTCH and GM-CSF-controlled manner., (© 2024. The Author(s).)

Background: Weaning protocols from Non-invasive ventilation (NIV) are scant. We set out to study a protocol that was different from the British Thoracic Society protocol and lay down a weaning protocol from NIV in patients with Acute acidotic hypercapnic respiratory failure (AAHRF)., Materials and Methods: Patients admitted with AAHRF and treated with NIV (baseline pH<7.35, PaCO2 >45 mmHg, and not requiring intubation) at a tertiary care teaching hospital, after taking into consideration the inclusion and the exclusion criteria were randomised in to one of the two group of weaning form NIV and serial ABGs were monitored., Results: The primary outcome of the study shows that there was no significant differences in the success rates of weaning from NIV in both the arms. The secondary outcome shows a few factors such as age, gender, SAPS2 score having an effect on the determination of weaning failure., Conclusion: Our study showed that both weaning by duration reduction and pressure reduction had equal success rates but a point on noting the SAPS II score on admission and the age of a particular patient will help decide on weaning initiation.CTRI/2019/12/022560 [Registered on: 30/12/2019]., (Copyright © 2024 Copyright: © 2024 Indian Chest Society.)

Objective: The aim of this study is to evaluate the presence of laryngopharyngeal reflux in patients with voice disorders thereby aiding in the early management and improving the quality of life., Study Design: Cross Sectional study., Methods: This cross sectional study was carried out in a tertiary care hospital, patients with history of voice change for more than 3 weeks were included, and divided into 4 groups depending upon the level of voice use. Patients were asked to fill Koufmann Reflux Symptom Index questionnaire followed by video laryngoscopy and findings were plotted according to Reflux Finding Score. Based on the scoring, impact of LPRD in patient with voice disorders was analysed., Results: Among the 90 study participants, 74 (82.2%) were found to have LPRD. The mean age was 42.76 ± 10.33 years. Majority (43.2%) belong to the age group of 41-50 years, with female predominance (70.3%). Majority (41.9%) of them were level IV voice users. 59.5% were positive Koufman reflex symptom index, 67.6% were positive Reflex finding score. Hoarseness (58.1%) the most common symptom., Conclusion: Laryngopharyngeal reflux disease has to be considered in patients presenting with hoarseness of voice for more than three weeks. Simple and highly reproducible scores like Reflux Symptom Index and Reflux Finding Score proven useful and valuable tools in diagnosing LPRD thereby aiding in early diagnosis and prompt management and improves the patient's quality of life., Level of Evidence: Level 1., Competing Interests: Conflict of InterestNot applicable., (© Association of Otolaryngologists of India 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Solitary fibrous tumors (SFTs) are rare neoplasms primarily found in the pleural region but have been documented in diverse extrapleural sites, including the nasal cavity and paranasal sinuses, albeit infrequently. Here, we present a case of a 48-year-old female who presented with a right-sided nasal mass and associated ophthalmologic symptoms, ultimately diagnosed with a benign spindle cell lesion localized to the nasal cavity. The patient underwent a comprehensive evaluation involving clinical examination, radiological imaging, and histopathological analysis, leading to the identification of a benign solitary fibrous tumor. Notably, diagnosing SFTs in the nasal cavity presents challenges due to their nonspecific clinical and imaging features, necessitating a multidisciplinary approach for accurate diagnosis and optimal management. Surgical excision, preferably via endoscopic techniques, remains the cornerstone of treatment based on tumor characteristics and extent. This case underscores the importance of recognizing uncommon presentations of sinonasal lesions, navigating diagnostic complexities, and emphasizing the critical role of multidisciplinary collaboration in achieving favorable treatment outcomes for patients with such nasal cavity tumors., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2024, Vijayasundaram et al.)

In the current study, the co-precipitation technique was employed for the synthesis of Cadmium oxide (CdO) and Copper‒doped Cadmium oxide (Cu‒CdO) nanoparticles. The synthesized samples were subjected to powder X-Ray diffraction (P-XRD), Field emission scanning electron microscopy (FE-SEM), Energy-dispersive X-ray (EDX), Fourier transforms Infrared (FT-IR), UV-Vis spectroscopy, photoluminescence (PL), laser-induced fluorescence spectroscopy and antibacterial investigations. According to the P-XRD analysis, both the samples were simple cubic in structure and have average grain sizes of 54 and 28 nm, respectively. FE-SEM was deployed to explore the surface textures of the samples. EDX technique was used to look at the elemental compositions of the samples. The technique of FT-IR was employed to identify the vibrational modes. UV-Vis spectra in diffuse reflectance mode were obtained and the optical bandgaps of the CdO and Cu‒CdO samples were obtained as 4.52 eV and 2.83 eV, respectively. The photoluminescence studies were conducted at an excitation wavelength of 300 nm and emission peaks were red-shifted in both samples. Fluorescence spectroscopy was applied to explore the lifetimes of synthesized nanoparticles. The technique of Agar-well diffusion was applied to assess the antibacterial performance of the generated nanoparticles against Micrococcus Luteus (gram-positive) and Escherichia coli (gram-negative) bacterium at variable concentrations. Both samples in the current study are significantly effective against both bacterial strains., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Background: Indwelling urethral catheters (IDC) are ubiquitous to healthcare settings, and are associated with many familiar risks like haematuria, infections, bladder spasms and stones. However, a less known complication is catheter-associated meatal pressure injury (CAMPI), especially in those with long-term IDCs. The objective of this study was to explore the prevalence, associated features and management of CAMPI in adults with a long-term IDC., Methods: A cross-sectional multi-centre study was undertaken of 200 adults with a long-term IDC across regional south-west Queensland, Australia between June 2019 to June 2021. The prevalence of CAMPI was determined by clinical examination, voluntary surveys completed by participants and documentation in medical records. Key IDC statistics included total duration of IDC, location of IDC changes, IDC size, type and fixation., Results: Out of 200 adults with a long-term IDC, 9% (18/200) had a CAMPI. There was a higher prevalence of male CAMPI (17/169, 10%) compared to female CAMPI (1/31, 3%). The median time to identification of a CAMPI after initial IDC insertion was 12 weeks (2-136 weeks), but occurred as soon as 2 weeks. CAMPI formation was associated with IDC changes in the community, impaired mobility and congestive cardiac failure (CCF). CAMPI were mostly treated by conservative means given the frailty of the population., Conclusions: Poor mobility, community-managed IDCs, and CCF were all found to have statistically significant associations with the development of CAMPI. CAMPI represents an important and underserved iatrogenic complication within urology practice, and greater awareness is needed to prevent it in vulnerable patients with long-term IDCs., Competing Interests: Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at https://tau.amegroups.com/article/view/10.21037/tau-23-445/coif). The authors have no conflicts of interest to declare., (2024 Translational Andrology and Urology. All rights reserved.)

The incorporation of sex and gender variables in medical education can be made more intuitive and accurate with thoughtful intervention. The authors propose a flowchart to make medical education tools more precise and applicable to patients who would benefit from a nuanced approach to sex and gender. The Body Guidelines can empower educators to consider sex and gender in terms of the etiology and epidemiology of any given pathologic process or disease., Competing Interests: The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (© The Author(s) 2023.)

Background and Purpose: Cervical Spondylotic Myelopathy (CSM) is a gradually escalating spinal cord disturbance set in motion by the degenerative narrowing of the vertebral canal. Routine MRI may fail to detect the subtle early alterations of the cord. MRI Diffusion Tensor Imaging (DTI) possesses the potential to detect these changes. This study intends to estimate the potential of the DTI technique in non-stenotic & stenotic spinal canals in individuals affected with CSM., Methodology: Sixty-four subjects who met the requirements of the inclusion criteria were incorporated into the investigation. All subjects underwent routine MRI sequences in addition to DTI of the cervical spine region. Scalars such as Fractional Anisotropy (FA), besides Apparent Diffusion Coefficient (ADC), were computed at each cervical intervertebral fibrocartilaginous disc level for all subjects. DTI fiber tractography was then performed to qualitatively assess the microstructural integrity of the tracts., Results: A noteworthy difference (p<0.05) was seen in the FA parameter and ADC parameter values between the stenotic and non-stenotic groups, with the non-stenotic group having a higher mean FA and a lower ADC than the stenotic group (at the level of stenosis). A significant difference in age was seen between both groups, with most of the patients in the stenotic group belonging to 40 years and above. Tractography helped in demonstrating the morphology of the fiber tracts., Conclusion: DTI parameters, namely FA and ADC, are sensitive to damage to the white matter and can be used to detect microstructural changes in the cord. However, standardization of the protocol is necessary when imaging the spinal canal., (Copyright © 2023. Published by Elsevier Inc.)

Background: 'Regenerative endodontics' using host-derived scaffolds and biomaterials (MTA) is popular in the management of teeth with open apex. Alkaline pH and bioactivity contribute to tissue healing and remineralization. We assessed the influence of PRF and Blood Clot on the pH and Ca + ion release from MTA. Methods: A total of 15 single-rooted human extracted teeth were sectioned at the level of the cementoenamel junction. Based on the type of scaffolds used, samples were divided into three groups. Group 1 (MTA+ PRF), Group 2 (MTA + Blood Clot), Group 3 (control MTA). The prepared specimens were transferred to a fresh falcon tube containing 10mL of distilled water and the collected solutions were analysed for pH and Ca + ion release at 3h, seventh day and 14 th day. Results: It was observed that the mean pH and Ca + ion release were significantly lower in the experimental groups as compared to the control group. Though there was an increase in the pH recorded in Group 1 and 2 at all time periods, the difference was not significant. Ca + ion release peaked at Day 7 (Group3 > Group2 > Group1) and reduced significantly on the 14 th day for all groups. Conclusions: Within the limitations of the study, it can be concluded that PRF and blood clot influence the pH and Ca + ion release from MTA., Competing Interests: No competing interests were disclosed., (Copyright: © 2023 Khatri S et al.)

Introduction: The genus Equisetum (Equisetaceae) is cosmopolitan in distribution, with 41 recognized species. Several species of Equisetum are widely used in treating genitourinary and related diseases, inflammatory and rheumatic problems, hypertension, and wound healing in traditional medicine practices worldwide. This review intends to present information on the traditional uses, phytochemical components, pharmacological activities, and toxicity of Equisetum spp. and to analyze the new insights for further study., Methods: Relevant literature has been scanned and collected via various electronic repositories, including PubMed, Science Direct, Google Scholar, Springer Connect, and Science Online, from 1960 to 2022., Results: Sixteen Equisetum spp. were documented as widely used in traditional medicine practices by different ethnic groups throughout the world. A total of 229 chemical compounds were identified from Equisetum spp. with the major group of constituents being flavonol glycosides and flavonoids. The crude extracts and phytochemicals of Equisetum spp. exhibited significant antioxidant, antimicrobial, anti-inflammatory, antiulcerogenic, antidiabetic, hepatoprotective, and diuretic properties. A wide range of studies have also demonstrated the safety of Equisetum spp., Conclusion: The reported pharmacological properties of Equisetum spp. support its use in traditional medicine, though there are gaps in understanding the traditional usage of these plants for clinical experiments. The documented information revealed that the genus is not only a great herbal remedy but also has several bioactives with the potential to be discovered as novel drugs. Detailed scientific investigation is still needed to fully understand the efficacy of this genus; hence, very few Equisetum spp. were studied in detail for phytochemical and pharmacological investigation. Moreover, its bioactives, structure-activity connection, in vivo activity, and associated mechanism of action ought to be explored further., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Investigating the RNA regulation landscape primarily relies on understanding how RNA-protein interactions are governed in various cell types, including neurons. Analysis of RNA-protein interactions in physiological environments warrants the development of new tools that rely on RNA manipulation. Recently, a CRISPR-based RNA-editing tool (dCas13b-ADAR2 DD ) was developed to mitigate disease-associated point mutations in cell lines. Here, we explored the targeted sequence editing potential of the tool (dCas13b-ADAR2 DD system) by adapting it to manipulate RNA function to visualize RNA editing in primary hippocampal neurons. This two-component system includes a programmable guide RNA (gRNA) complementary to the target RNA and a catalytically dead version of the Cas13b enzyme fused to ADAR. The RNA editing protocol outlined in this article relies on gRNA-dependent targeting of the dCas13b-ADAR fusion protein to the mutant form of the Dendra2 transcript. Dendra2 is not required for intrinsic cellular functioning. It was ectopically expressed for fluorescent detection as a proof-of-principle demonstration of targeted RNA editing. We first abrogated the fluorescence of Dendra2 by introducing a nonsense mutation that precludes the formation of the functional protein. To visualize the efficacy of the RNA editing in neurons, we used the dCas13b-ADAR2 DD system to edit specific nucleotides within the Dendra2 mRNA to restore the amino acid codes critical for Dendra2 fluorescence. This method lays the foundation for future studies on the dynamics of activity-induced RNA-protein interactions in neurons and can be extended to manipulate the endogenous RNome in diverse neuronal subtypes. Furthermore, this methodology will enable investigators to visualize the spatial and temporal resolution of RNA-protein interactions without altering the genomes via conventional methods. © 2023 Wiley Periodicals LLC. Support Protocol: Preparation of mouse primary hippocampal culture Basic Protocol: Targeted editing of RNA., (© 2023 Wiley Periodicals LLC.)

The considerable improvement of technology produced for various applications has resulted in a growth in data sizes, such as healthcare data, which is renowned for having a large number of variables and data samples. Artificial neural networks (ANN) have demonstrated adaptability and effectiveness in classification, regression, and function approximation tasks. ANN is used extensively in function approximation, prediction, and classification. Irrespective of the task, ANN learns from the data by adjusting the edge weights to minimize the error between the actual and predicted values. Back Propagation is the most frequent learning technique that is used to learn the weights of ANN. However, this approach is prone to the problem of sluggish convergence, which is especially problematic in the case of Big Data. In this paper, we propose a Distributed Genetic Algorithm based ANN Learning Algorithm for addressing challenges associated with ANN learning for Big data. Genetic Algorithm is one of the well-utilized bio-inspired combinatorial optimization methods. Also, it is possible to parallelize it at multiple stages, and this may be done in an extremely effective manner for the distributed learning process. The proposed model is tested with various datasets to evaluate its realizability and efficiency. The results obtained from the experiments show that after a specific volume of data, the proposed learning method outperformed the traditional methods in terms of convergence time and accuracy. The proposed model outperformed the traditional model by almost 80% improvement in computational time., (© 2023. The Author(s).)

Here, we report the genome sequence of a Pasteurella multocida strain isolated from the heart blood of a spotted deer (Bareilly, India). The 2.44-Mbp genome has 2,227 coding sequences, with a G+C content of 40.7%., Competing Interests: The authors declare no conflict of interest.

Purpose: To assess the negative predictive value of PSMA PET scan for lymph node staging in patients undergoing robotic radical prostatectomy and pelvic lymph node dissection., Materials and Methods: A retrospective analysis of patients who underwent robotic-assisted radical prostatectomy with pelvic lymph node dissection and had a preoperative negative PSMA PET scan for metastasis was performed. The documented pre-operative variables studied included age, BMI, PSA at diagnosis, Gleason score, and biopsy ISUP grades. Patients were categorised as low, intermediate and high risk according to the D Amico classification. The post-op variables included were number of lymph nodes harvested, number of positive nodes, positivity rate, size of the node metastasis, T staging and ISUP grading., Results: The overall negative predictive value of PSMA PET scan was 71.6%. Further sub-classification according to risk stratification demonstrated a NPV of 58.02%, 92.7% and 90% for high, intermediate and low risk, respectively., Conclusion: Pelvic lymph node dissection cannot be excluded based on a negative preop PSMA PET/CT scan., (© 2023. The Author(s), under exclusive licence to Springer Nature B.V.)

In vitro transcription (IVT) is a DNA-templated process for synthesizing long RNA transcripts, including messenger RNA (mRNA). For many research and commercial applications, IVT of mRNA is typically performed using bacteriophage T7 RNA polymerase (T7 RNAP) owing to its ability to produce full-length RNA transcripts with high fidelity; however, T7 RNAP can also produce immunostimulatory byproducts such as double-stranded RNA that can affect protein expression. Such byproducts require complex purification processes, using methods such as reversed-phase high-performance liquid chromatography, to yield safe and effective mRNA-based medicines. To minimize the need for downstream purification processes, we rationally and computationally engineered a double mutant of T7 RNAP that produces substantially less immunostimulatory RNA during IVT compared with wild-type T7 RNAP. The resulting mutant allows for a simplified production process with similar mRNA potency, lower immunostimulatory content and quicker manufacturing time compared with wild-type T7 RNAP. Herein, we describe the computational design and development of this improved T7 RNAP variant., (© 2022. The Author(s).)

Molecular docking (Mol.Doc) techniques were employed to ascertain the binding affinity of two resorcinol-based acridinedione dyes (ADR1 and ADR2) with the widely studied globular protein Bovine Serum Albumin (BSA) in the presence of site-selective binding drugs by Autodock Vina 4.2 software. Docking of various feasible conformers of ADR1 dye with BSA was found to be energetically more favored than ADR2 dye, even though both these dyes differ in the 9th position of the basic dye structure. Analysis of dyes with BSA establishes the location of dye in all of the binding sites of BSA, predominantly through conventional and nonconventional hydrogen-bonding (HB) interactions. The coexistence of hydrophobic interactions resulted in the stability of various conformers generated. The introduction of site I and site II (Sudlow site binding drugs) into ADR1-BSA and ADR2-BSA complexes effectively destabilizes the dye-protein complex; however, the drugs do not displace ADR dyes completely from their selective binding domains. Site II binding drugs effectively destabilize the binding ability of the dye-protein complex rather than site I drugs. However, docking of site I drug 3-carboxyl-4-methyl-5-propyl-2-furanpropanic acid (CMPF) largely destabilizes the ADR1-protein complex, whereas indomethacin (INDO) enhances the binding affinity of the ADR2-protein complex. Interestingly, simultaneous docking of ADR dyes to the BSA-drug complex results in larger stability of the protein-drug complex through HB interactions rather than hydrophobic interactions. Both ADR1 and ADR2 dyes predominantly occupy the Sudlow binding sites of BSA, and the introduction of either site I or site II binding drugs does not displace the dye efficiently from the corresponding binding sites, rather the drugs are effectively displaced toward other binding domains apart from their specific site-binding domains of BSA. Through Mol.Doc techniques, we authenticate that the interactions in host-guest complex systems involving competing ligands are established in depth, wherein the dye as well as the amino acid (AA) moieties in BSA act as both HB donor and acceptor sites apart from several hydrophobic interactions coexisting toward the stability., Competing Interests: The authors declare no competing financial interest., (© 2023 The Authors. Published by American Chemical Society.)

Enzyme-coupled titanium oxide nanopowder samples were prepared usingdifferent volumes of vermiwash using a cost-effective soft chemical method and their photocatalytic efficiency was studied against Methylene Blue (MB) dye decomposition. The volume of vermiwash used in the starting solution was varied from 50 to 200 mL in steps of 50 mL and the effect of enzymes prevalent in the vermiwash on the photocatalytic activity of titanium oxide (TiO 2 ) was studied. The resultant water obtained after the photocatalytic dye degradation was found to inherit the enzymes from the nanoproduct. This enzyme-activated treated water showed effective seed germination of black gram (Vigna mungo L.). The results suggested that the enzyme-coupled TiO 2 can be used as an effective and eco-friendly material for the treatment of contaminated water and consequently the treated water can also be utilized for enhanced seed germination., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: K. Ravichandran reports financial support was provided by Science and Engineering Research Board. K. Ravichandran reports financial support was provided by DST-FIST., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Background: Panx1 (pannexin 1) forms high conductance channels that secrete ATP upon stimulation. The role of Panx1 in mediating constriction in response to direct sympathetic nerve stimulation is not known. Additionally, it is unknown how the expression level of Panx1 in smooth muscle cells (SMCs) influences α-adrenergic responses. We hypothesized that the amount of Panx1 in SMCs dictates the levels of sympathetic constriction and blood pressure., Methods: To test this hypothesis, we used genetically modified mouse models enabling expression of Panx1 in vascular cells to be varied. Electrical field stimulation on isolated arteries and blood pressure were assessed., Results: Genetic deletion of SMC Panx1 prevented constriction by electric field stimulation of sympathetic nerves. Conversely, overexpression of Panx1 in SMCs using a ROSA26 transgenic model increased sympathetic nerve-mediated constriction. Connexin 43 hemichannel inhibitors did not alter constriction. Next, we evaluated the effects of altered SMC Panx1 expression on blood pressure. To do this, we created mice combining a global Panx1 deletion, with ROSA26-Panx1 under the control of an inducible SMC specific Cre (Myh11). This resulted in mice that could express only human Panx1, only in SMCs. After tamoxifen, these mice had increased blood pressure that was acutely decreased by the Panx1 inhibitor spironolactone. Control mice genetically devoid of Panx1 did not respond to spironolactone., Conclusions: These data suggest Panx1 in SMCs could regulate the extent of sympathetic nerve constriction and blood pressure. The results also show the feasibility humanized Panx1-mouse models to test pharmacological candidates.

The Retention Using Selective Hooks (RUSH) system allows for the synchronized release of one or more proteins of interest from a donor endomembrane compartment, usually the endoplasmic reticulum, and the subsequent monitoring of their traffic toward acceptor compartments. Here we describe the RUSH system applied to cytotoxic T cells to characterize the biogenesis of lytic granules, using as a proof-of-concept granzyme B trafficking to this specialized compartment., (© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Since the workplace has become desk-based and leisure time has become dominated by digital relaxation modes, the world is moving at a pace where physical activity has become a time-bound routine. The negative effects of extended sitting are a global concern since the workforce is becoming more desk based. There is a dearth of reviews that may link the knowledge on the effects of sedentary behaviour on hypertension and its accompanying damage to the brain and blood vessels and provide a future scope for the investigations connected to the relationship between sedentary behaviour and hypertension. Methods: Based on the database search and extensive research we did, we found studies that concentrated on the adverse effects of sedentary behaviour in association with blood pressure, cognitive decline and brain damage on adults. Results: We extracted 12 articles out of 20,625. We identified the potential adverse effects of sedentary behaviour, methods to reduce sedentary behaviour and the positive changes on health due to the interventions introduced. Sedentary lifestyle has shown a decline in human health. However, the visible symptoms presented later in life makes it very important to know the areas of decline and look for ways to curb the decline or procrastinate it.

Background: Atherosclerosis being the keystone in the pathology of coronary artery disease (CAD) is a chronic inflammation of arterial intima mediated by various inflammatory markers. Pentraxin 3 (PTX3) and high-sensitivity C-reactive protein (hs-CRP) are the two important biomarkers of chronic inflammation that causes atherosclerosis., Aims: This study aims to investigate the association of serum PTX3 and hs-CRP with the severity of coronary stenosis in patients undergoing coronary angiogram., Subjects and Methods: A total of 80 patients who underwent elective coronary angiogram were included. Their blood sample was collected for PTX3 and hs-CRP estimation prior to angiogram. Based on the angiogram, the participants were divided into four groups based on the number of arteries affected. PTX3 was estimated using enzyme-linked immunosorbent assay and hs-CRP was assayed using latex-enhanced immunosorbent assay., Statistical Analysis Used: Kruskal-Wallis test was used to find the association of PTX3 and hs-CRP in each group and Pearson's correlation was used to correlate PTX3 and hs-CRP with the extent of stenosis., Results: The mean PTX3 and hs-CRP levels in patients with some lesions in the coronary artery were 231.5 ± 129.9 pg/mL and 2.4 ± 0.4 mg/mL, respectively. The PTX3 levels elevate gradually with the severity of stenosis with P = 0.000 which is highly significant. A strong positive correlation was observed ( R = 0.7929, P < 0.00001) with PTX3 and severity of stenosis. Whereas, for hs-CRP, the correlation was weaker ( R = 0.3011, P = 0.006)., Conclusions: PTX3 and hs-CRP can not only predict the number of arteries affected but also can differentiate between normal coronaries and CAD which can minimize the use of angiography., Competing Interests: There are no conflicts of interest., (Copyright: © 2022 International Journal of Applied and Basic Medical Research.)

Herein, we assessed the stage-specific efficacy of inositol hexaphosphate (IP6, phytic acid), a bioactive food component, on prostate cancer (PCa) growth and progression in a transgenic mouse model of prostate cancer (TRAMP). Starting at 4, 12, 20, and 30 weeks of age, male TRAMP mice were fed either regular drinking water or 2% IP6 in water for ~8-15 weeks. Pathological assessments at study endpoint indicated that tumor grade is arrested at earlier stages by IP6 treatment; IP6 also prevented progression to more advanced forms of the disease (~55-70% decrease in moderately and poorly differentiated adenocarcinoma incidence was observed in advanced stage TRAMP cohorts). Next, we determined whether the protective effects of IP6 are mediated via its effect on the expansion of the cancer stem cells (CSCs) pool; results indicated that the anti-PCa effects of IP6 are associated with its potential to eradicate the PCa CSC pool in TRAMP prostate tumors. Furthermore, in vitro assays corroborated the above findings as IP6 decreased the % of floating PC-3 prostaspheres (self-renewal of CSCs) by ~90%. Together, these findings suggest the multifaceted chemopreventive-translational potential of IP6 intervention in suppressing the growth and progression of PCa and controlling this malignancy at an early stage.

A variety of intrinsic and extrinsic factors contribute to the altered efficiency of CTLs in elderly organisms. In particular, the efficacy of antiviral CD8 + T cells responses in the elderly has come back into focus since the COVID-19 pandemic outbreak. However, the exact molecular mechanisms leading to alterations in T cell function and the origin of the observed impairments have not been fully explored. Therefore, we investigated whether intrinsic changes affect the cytotoxic ability of CD8 + T cells in aging. We focused on the different subpopulations and time-resolved quantification of cytotoxicity during tumor cell elimination. We report a surprising result: Killing kinetics of CD8 + T cells from elderly mice are much faster than those of CD8 + T cells from adult mice. This is true not only in the total CD8 + T cell population but also for their effector (T EM ) and central memory (T CM ) T cell subpopulations. TIRF experiments reveal that CD8 + T cells from elderly mice possess comparable numbers of fusion events per cell, but significantly increased numbers of cells with granule fusion. Analysis of the cytotoxic granule (CG) content shows significantly increased perforin and granzyme levels and turns CD8 + T cells of elderly mice into very efficient killers. This highlights the importance of distinguishing between cell-intrinsic alterations and microenvironmental changes in elderly individuals. Our results also stress the importance of analyzing the dynamics of CTL cytotoxicity against cancer cells because, with a simple endpoint lysis analysis, cytotoxic differences could have easily been overlooked., (© 2022 The Authors. Aging Cell published by Anatomical Society and John Wiley & Sons Ltd.)

Background: Despite constant advances in science, obscurity remains in the efficient removal of pulp stones to aid in successful root canal treatment. In this context, chemical means of dissolving pulp stones were explored., Aim: The aim of this study is to evaluate and to compare the efficacy of decalcifying agents on the dissolution of pulp stones., Materials and Methods: The study was divided into two groups for pulp stone analysis (21 samples) and dentin analysis (54 samples). Twenty-one pulp stones from patients aged 18-70 who underwent root canal treatment were collected and divided into three subgroups ( n = 7) randomly. They were subjected to chemical treatment in a labeled glass container with 5 ml of the respective chemical agents, such as 17% ethylenediaminetetraacetic acid solution (positive control), no treatment (negative control), and newly developed Physiological Simulated Decalcifying Agent (PSDA). At the end of the study period (24 h), the samples were removed, rinsed with deionized water, and subjected to physical analysis, scanning electron microscopy (SEM), and Energy -dispersive X-ray spectroscopy (EDS) analysis. Under dentin analysis, 54 maxillary premolars scheduled for orthodontic extraction without caries or extensive restorations were selected, following which 2-mm thick transverse dentinal sections at the cementoenamel junction level were obtained and randomly divided into two groups for SEM ( n = 21) and microhardness analysis ( n = 33). The samples were subjected to respective chemical treatment groups similar to pulp stones for 24 h and analyzed using SEM, EDS, and microhardness analysis., Results: Postchemical treatment with the newly developed decalcifying solution, the pulp stones showed the absence of nodular crystallites and surface softening under SEM and a decrease in the calcium level under EDS analysis. Concerning the microhardness of dentin, no significant changes could be observed., Conclusion: The newly explored PSDA was found to be efficacious in the decalcification of pulp stones at a clinically relevant time of 24 h, without significantly affecting the structural integrity and the hardness values of dentin., Competing Interests: There are no conflicts of interest., (Copyright: © 2022 Journal of Conservative Dentistry.)

SARS-CoV-2 cause fatal infection in 213 countries accounting for the death of millions of people globally. In the present study, phytochemicals from spices were assessed for their ability to interact with SARS-CoV-2 M Pro . Structure based virtual screening was performed with 146 phytochemicals from spices using Autodock Vina. Phytochemicals with binding energy ≥ -8.0 kcal/mol were selected to understand their interaction with M Pro . Virtual screening was further validated by performing molecular docking to generate favorable docked poses and the participation of important amino acid residues. Molecular dynamics simulation for the docked poses was performed to study thermodynamic properties of the protein, ligand and protein-ligand complexes. The finding shows that cinnamtannin B2 and cyanin showed favorable binding affinity values with SARS-CoV-2 M Pro . The results are comparable in terms of docked poses, important amino acid participation and thermodynamic properties with the standard control drugs remdesivir, benazepril and hydroxychloroquine diphosphate. Prime MM-GBSA was employed for end-point binding energy calculation. Binding to domain I and II of M Pro were mediated through the OH, SH, NH 2 and non-polar side chain of amino acids. Cinnamtannin B2 and cyanin binds to M Pro with many sub sites within the active site with RMSD and RMSF within 4 Å. The results computed using Prime MM-GBSA show that cinnamtannin B2 (-68.54940214 kcal/mol) and cyanin (-62.1902835 kcal/mol) have better binding affinity in comparison to hydroxychloroquine diphosphate (-54.00912412 kcal/mol) and benazepril (-53.70242369 kcal/mol). The results provide a basis for exploiting cinnamtannin B2 and cyanin as a starting point potential candidate for the development of drug against SARS-CoV-2.Communicated by Ramaswamy H. Sarma.

Cytotoxic T lymphocytes (CTL) kill malignant and infected cells through the directed release of cytotoxic proteins into the immunological synapse (IS). The cytotoxic protein granzyme B (GzmB) is released in its soluble form or in supramolecular attack particles (SMAP). We utilize synaptobrevin2-mRFP knock-in mice to isolate fusogenic cytotoxic granules in an unbiased manner and visualize them alone or in degranulating CTLs. We identified two classes of fusion-competent granules, single core granules (SCG) and multi core granules (MCG), with different diameter, morphology and protein composition. Functional analyses demonstrate that both classes of granules fuse with the plasma membrane at the IS. SCG fusion releases soluble GzmB. MCGs can be labelled with the SMAP marker thrombospondin-1 and their fusion releases intact SMAPs. We propose that CTLs use SCG fusion to fill the synaptic cleft with active cytotoxic proteins instantly and parallel MCG fusion to deliver latent SMAPs for delayed killing of refractory targets., (© 2022. The Author(s).)

Antibiotic-resistant Neisseria gonorrhoeae (Ng ) are an emerging public health threat due to increasing numbers of multidrug resistant (MDR) organisms. We identified two novel orally active inhibitors, PTC-847 and PTC-672, that exhibit a narrow spectrum of activity against Ng including MDR isolates. By selecting organisms resistant to the novel inhibitors and sequencing their genomes, we identified a new therapeutic target, the class Ia ribonucleotide reductase (RNR). Resistance mutations in Ng map to the N-terminal cone domain of the α subunit, which we show here is involved in forming an inhibited α 4 β 4 state in the presence of the β subunit and allosteric effector dATP. Enzyme assays confirm that PTC-847 and PTC-672 inhibit Ng RNR and reveal that allosteric effector dATP potentiates the inhibitory effect. Oral administration of PTC-672 reduces Ng infection in a mouse model and may have therapeutic potential for treatment of Ng that is resistant to current drugs., Competing Interests: JN, SL, SJ, AG, JW, MA, GC, JH, MD, GK, AB was employed by PTC Therapeutics when the work was performed and received salary and compensation during their tenure, KR, TL, CC, CD, JS No competing interests declared, (© 2022, Narasimhan et al.)

Enhancing the potency of mRNA therapeutics is an important objective for treating rare diseases, since it may enable lower and less-frequent dosing. Enzyme engineering can increase potency of mRNA therapeutics by improving the expression, half-life, and catalytic efficiency of the mRNA-encoded enzymes. However, sequence space is incomprehensibly vast, and methods to map sequence to function (computationally or experimentally) are inaccurate or time-/labor-intensive. Here, we present a novel, broadly applicable engineering method that combines deep latent variable modelling of sequence co-evolution with automated protein library design and construction to rapidly identify metabolic enzyme variants that are both more thermally stable and more catalytically active. We apply this approach to improve the potency of ornithine transcarbamylase (OTC), a urea cycle enzyme for which loss of catalytic activity causes a rare but serious metabolic disease., (© 2022. The Author(s).)

In the title compound [systematic name: 1-(3-methyl-2,6-diphenylpiperidin-1-yl)ethanone], C 20 H 23 NO, the piperidine ring adopts a distorted boat conformation, while the phenyl rings subtend a dihedral angle 65.1 (2)°. In the crystal, mol-ecules are linked by C-H⋯O hydrogen bonds into chains extending along the b- axis direction. The DFT/B3LYP/6-311 G(d,p) method was used to determine the HOMO-LUMO energy levels. A Hirshfeld surface analysis was conducted to verify the contributions of the different inter-molecular inter-actions, indicating that the important contributions to the crystal packing are from H⋯H (73.2%), C⋯H (18.4%) and O⋯H (8.4%) inter-actions., (© Periyannan et al. 2022.)

The role of cloud services in the data-intensive industry is indispensable. Cision recently reported that the cloud market would grow to 55 billion USD, with an active contribution of the cloud to healthcare around 2025. Inspired by the report, cloud vendors expand their market and the quality of services to seek growth globally. The rapid growth of the cloud sector in the healthcare industry imposes a challenge: making a rational choice of a cloud vendor (CV) out of a diverse set of vendors. Typically, the healthcare industry 4.0 sees the issue as a large-scale group decision-making problem. Previous studies on a CV selection face certain challenges, such as (i) a lack of the ability to handle multiple users' views, as well as experts'/users' complex linguistic views; (ii) the confidence level associated with a view is not considered; (iii) the transformation of multiple users' views into holistic data is lacking; and (iv) the systematic prioritization of CVs with minimum human intervention is a crucial task. Motivated by these challenges and circumventing them, a new big data-driven decision model is put forward in this paper. Initially, the data in the form of complex expressions are collected from multiple cloud users and are further transformed into a holistic decision matrix by adopting probabilistic linguistic information (PLI). PLI represents complex linguistic expressions along with the associated confidence levels. Later, a holistic decision matrix is formed with the missing values imputed by proposing an imputation algorithm. Furthermore, the criteria weights are determined by using a newly proposed mathematical model and partial information. Finally, the evaluation based on the distance from average solution (EDAS) approach is extended to PLI for the rational ranking of CVs. A real-time example of a CV selection for a healthcare center in India is exemplified so as to demonstrate the usefulness of the model, and the comparison reveals the merits and limitations of the model., Competing Interests: Conflict of interestAll authors of this research paper declare that, there is no conflict of interest., (© The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature 2021.)

Coronavirus disease (COVID-19) caused by SARS-CoV-2 was notified from Wuhan city, Hubei province, China in the mid of December 2019. The disease is showing dynamic change in the pattern of confirmed cases and death toll in these low and middle-income countries (LMICs). In this study, exponential growth (EG) method was used to calculate the real-time reproductive number (R t ) for initial and later stage of epidemic in South Asian Association for Regional Cooperation (SAARC) member countries (April 2020 - December 2020). Time dependent (TD) method was used to calculate the weekly real -time reproduction number (R t ). We also presented the observations on COVID-19 epidemiology in relation with the health expenditure, poverty, BCG vaccination, literacy population density and R t for understanding the current scenario, trends, and expected outcome of the disease in SAARC countries. A significant positive correlation was noticed between COVID-19 deaths and health expenditure (% GDP) (r = 0.58, P  < 0.05). The other factors such as population density/sq km, literacy %, adult population %, and poverty % were not significantly correlated with number of COVID-19 cases and deaths. Among SAARC countries, the highest R t was observed in India (R t  = 2.10; 95% CI 2.04-2.17) followed by Bangladesh (R t  = 1.62; 95% CI 1.59-1.64) in initial state of epidemic. A continuous monitoring is necessitated in all countries looking at the medical facilities, available infrastructure and healthcare manpower, constraints which may appear with increased number of critically ill patients if the situation persists longer., Competing Interests: The authors declare that there are no conflicts of interest., (© 2021 Elsevier Ltd. All rights reserved.)

OTULIN is a deubiquitinase that specifically cleaves linear ubiquitin chains. Here we demonstrate that the ablation of Otulin selectively in keratinocytes causes inflammatory skin lesions that develop into verrucous carcinomas. Genetic deletion of Tnfr1, knockin expression of kinase-inactive Ripk1 or keratinocyte-specific deletion of Fadd and Mlkl completely rescues mice with OTULIN deficiency from dermatitis and tumorigenesis, thereby identifying keratinocyte cell death as the driving force for inflammation. Single-cell RNA-sequencing comparing non-lesional and lesional skin reveals changes in epidermal stem cell identity in OTULIN-deficient keratinocytes prior to substantial immune cell infiltration. Keratinocytes lacking OTULIN display a type-1 interferon and IL-1β response signature, and genetic or pharmacologic inhibition of these cytokines partially inhibits skin inflammation. Finally, expression of a hypomorphic mutant Otulin allele, previously shown to cause OTULIN-related autoinflammatory syndrome in humans, induces a similar inflammatory phenotype, thus supporting the importance of OTULIN for restraining skin inflammation and maintaining immune homeostasis., (© 2021. The Author(s).)

Obesity-induced inflammation is a major driving force in the development of insulin resistance, type 2 diabetes (T2D), and related metabolic disorders. During obesity, macrophages accumulate in the visceral adipose tissue, creating a low-grade inflammatory environment. Nuclear factor κB (NF-κB) signaling is a central coordinator of inflammatory responses and is tightly regulated by the anti-inflammatory protein A20. Here, we find that myeloid-specific A20-deficient mice are protected from diet-induced obesity and insulin resistance despite an inflammatory environment in their metabolic tissues. Macrophages lacking A20 show impaired mitochondrial respiratory function and metabolize more palmitate both in vitro and in vivo. We hypothesize that A20-deficient macrophages rely more on palmitate oxidation and metabolize the fat present in the diet, resulting in a lean phenotype and protection from metabolic disease. These findings reveal a role for A20 in regulating macrophage immunometabolism., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 The Author(s). Published by Elsevier Inc. All rights reserved.)

Objective: To determine breastmilk sodium changes in the first 72 hours after birth and to correlate maternal and neonatal variables with maternal breastmilk sodium., Methods: We enrolled 245 mothers and their exclusively breast-fed neonates in this prospective cohort study. Singleton, inborn babies of greater than 34 weeks gestation, who were exclusively breastfed for the first 72 hours were included. Babies who required neonatal intensive care unit (NICU) admission, top up feeds or discharged before 72 hours were excluded. Study outcomes were changes in breast milk sodium in the first 72 hours and association of high maternal breast milk sodium with various maternal and neonatal variables., Results: Mean (SD) breastmilk sodium steadily declined over the first 72 hours [53.5 (19.2), 38.5 (19.0) and 22.2 (10.6) mmol/L at 24, 48, 72 hours, respectively]. Breastfeeding £8 times per 24 hours in the first three days was the only factor significantly associated with high breastmilk sodium (P=0.008). Maternal age, gravida, mode of delivery, significant neonatal weight loss, hypernatremia, neonatal morbidities like fever, irritability, lethargy and poor suck had no significant correlation with high breastmilk sodium., Conclusions: Breastmilk sodium shows a steady decline in the first 72 hours after delivery. Feeding ≤8 times per day is associated high breastmilk sodium at 72 hours of age.

Avian chlamydiosis is one of the important neglected diseases with critical zoonotic potential. Chlamydia psittaci, the causative agent, affects most categories of birds, livestock, companion animals, and humans. It has many obscured characters and epidemiological dimensions, which makes it unique among other bacterial agents. Recent reports on transmission from equine to humans alarmed the public health authorities, and it necessitates the importance of routine screening of this infectious disease. High prevalence of spill-over infection in equines was associated with reproductive losses. Newer avian chlamydial species are being reported in the recent years. It is a potential biological warfare agent and the disease is an occupational hazard mainly to custom officers handling exotic birds. Prevalence of the disease in wild birds, pet birds, and poultry causes economic losses to the poultry industry and the pet bird trade. Interestingly, there are speculations on the 'legal' and 'illegal' bird trade that may be the global source of some of the most virulent strains of this pathogen. The mortality rate generally ranges from 5 to 40% in untreated cases, but it can sometimes be higher in co-infection. The intracellular lifestyle of this pathogen makes the diagnosis more complicated and there is also lack of accurate diagnostics. Resistance to antibiotics is reported only in some pathogens of the Chlamydiaceae family, but routine screening may assess the actual situation in all pathogens. Due to the diverse nature of the pathogen, the organism necessitates the One Health partnerships to have complete understanding. The present review focuses on the zoonotic aspects of avian chlamydiosis with its new insights into the pathogenesis, transmission, treatment, prevention, and control strategies. The review also briefs on the basic understandings and complex epidemiology of avian chlamydiosis, highlighting the need for research on emerging one health perspectives., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

Diabetic nephropathy (DN) is the most common manifestation of high glucose induced diabetes mellitus. In this study, we report the effects of Cassia auriculata ethanol leaf extract (CALE) on DN-associated cell toxicity and complications. The effects of CALE were screened in vitro using RGE cells. Cell viability was assessed using MTT and flow cytometry. Male Sprague-Dawley rats were divided into control, DN and treatment groups (n = 8). The DN and treatment groups received 60 mg/kg/bw of streptozotocin in citrate buffer, while the treatment group was administered 150 mg/kg/bw of CALE for 10 weeks. Biochemical analysis was conducted using spectrophotometry. Kidney tissues were analyzed using hematoxylin and eosin staining and transmission electron microscopy. CD365-KIM-1 expression was assessed using flow cytometry and signalling proteins were detected using western blotting. Treatment with 30-mM glucose reduced the viability of RGE cells in a time-dependent manner and increased the population of dead RGE cells. Cotreatment with CALE reduced cell death and glucose induced protein expression of LC3-II, RIP-1 and RIP-3 in a dose-dependent manner. In addition, CALE improved the biochemical complications, renal dysfunction and pathophysiology of rats with DN and partially or fully restored the expression of key DN-associated signalling proteins, such as KIM-1 LC3-II, RIP-1, RIP-3 and p-p38MAPK in kidney cells. CALE showed protective effects, and improved DN-associated complications in RGE cells under high glucose stress conditions, potentially by inhibiting autophagic-necroptosis signals. Additionally, CALE improved the biochemical and pathological features of kidney injury while reducing autophagic-necroptosis in rat renal cells via the LC3-II-RIP-p38MAPK pathway. PRACTICAL APPLICATIONS: Results from the current investigation will add information to the literature on glucose induced renal toxicity and the protective effects of CALE over the complications of diabetic nephropathy (DN). The mechanistic investigations of the study will add light on the autophagic/necroptosis signals in DN and open new routes of investigations to study the efficacy of CALE in diabetes-related complications., (© 2021 Wiley Periodicals LLC.)

The whole world is presently under threat from Coronavirus Disease 2019 (COVID-19), a new disease spread by a virus of the corona family, called a novel coronavirus. To date, the cases due to this disease are increasing exponentially, but there is no vaccine of COVID-19 available commercially. However, several antiviral therapies are used to treat the mild symptoms of COVID-19 disease. Still, it is quite complicated and uncertain decision to choose the best antiviral therapy to treat the mild symptom of COVID-19. Hesitant Fuzzy Sets (HFSs) are proven effective and valuable structures to express uncertain information in real-world issues. Therefore, here we used the hesitant fuzzy decision-making (DM) method. This study has chosen five methods or medicines to treat the mild symptom of COVID-19. These alternatives have been ranked by seven criteria for choosing an optimal method. The purpose of this study is to develop an innovative Additive Ratio Assessment (ARAS) approach to elucidate the DM problems. Next, a divergence measure based procedure is developed to assess the relative importance of the criteria rationally. To do this, a novel divergence measure is introduced for HFSs. A case study of drug selection for COVID-19 disease is considered to demonstrate the practicability and efficacy of the developed idea in real-life applications. Afterward, the outcome shows that Remdesivir is the best medicine for patients with mild symptoms of the COVID-19. Sensitivity analysis is presented to ensure the permanence of the introduced framework. Moreover, a comprehensive comparison with existing models is discussed to show the advantages of the developed framework. Finally, the results prove that the introduced ARAS approach is more effective and reliable than the existing models., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2021 Elsevier B.V. All rights reserved.)

IGF2 is essential in breast differentiation, lactation, tumor growth, and in breast cancer (BC) development and progression. This growth factor also inhibits apoptosis and promotes metastasis and chemoresistance, contributing to more aggressive tumors. We previously demonstrated that IGF2 protein levels are higher in BC tissues from African American women than in Caucasian women. We also showed that high IGF2 protein levels are expressed in normal breast tissues of African American women while little or no IGF2 was detected in tissues from Caucasian women. Others showed that decreased DNA methylation of the IGF2 gene leads to different BC clinical features. Thus, we designed this study to determine if differentially methylated regions of the IGF2 gene correspond to IGF2 protein expression in paired (Normal/Tumor) breast tissues and in BC cell lines. Methylation analysis was performed using Sodium Bisulphite Analysis and Methylation Sensitive Restriction Enzyme digestion methods. Our results show that a unique site in the INS-IGF2 region is hypermethylated in normal breast and hypomethylated in breast cancer. We designated this region the DVDMR. Furthermore, the methylation levels in the DVDMR significantly correlated with IGF2 protein levels. This novel DMR consists of 257bp localized in the INS-IGF2 gene. We propose that methylation of DVDMR represents a novel epigenetic biomarker that determines the levels of IGF2 protein expression in breast cancer. Since IGF2 promotes metastasis and chemoresistance, we propose that IGF2 levels contribute to BC aggressiveness. Validation of IGF2 as a biomarker will improve diagnosis and treatment of BC patients., Competing Interests: CONFLICTS OF INTEREST The authors declare no conflict of interest., (Copyright: © 2020 Radhakrishnan et al.)

Competing Interests: There are no conflicts of interest.

Objective: The objective of this study is to determine the clinical predictors of unsatisfactory Pap smears., Methodology: This was a case-control study done in a tertiary care institute. All unsatisfactory conventional pap (CP) smears between January 2015 and June 2017 were retrieved, and the slides were viewed. Clinical details were recorded from request forms and case files. Simple and multiple logistic regression analyses were used to identify the predictors of unsatisfactory CP smears., Results: In this study, we have included 314 unsatisfactory Pap smears and 541 controls with satisfactory Pap smears. Clinical parameters such as older age and cervical erosion proved to be important predictors of unsatisfactory pap smears. The most common reason for unsatisfactory pap smears was due to a paucity of epithelial elements (66.6%), followed by obscuration of smear details by blood/inflammatory cells/mucus (9.9%) and air drying artifacts (4.4%). There were multiple reasons in 19.1% of cases with unsatisfactory pap smears., Conclusion: Our study shows that older age groups and cervical erosion are predictors of unsatisfactory pap smears. Incidence of unsatisfactory pap smears can be reduced by education and retraining of health-care workers and doctors., Competing Interests: There are no conflicts of interest., (Copyright: © 2021 Journal of Mid-life Health.)

Understanding T cell function in vivo is of key importance for basic and translational immunology alike. To study T cells in vivo, we developed a new knock-in mouse line, which expresses a fusion protein of granzyme B, a key component of cytotoxic granules involved in T cell-mediated target cell-killing, and monomeric teal fluorescent protein from the endogenous Gzmb locus. Homozygous knock-ins, which are viable and fertile, have cytotoxic T lymphocytes with endogeneously fluorescent cytotoxic granules but wild-type-like killing capacity. Expression of the fluorescent fusion protein allows quantitative analyses of cytotoxic granule maturation, transport and fusion in vitro with super-resolution imaging techniques, and two-photon microscopy in living knock-ins enables the visualization of tissue rejection through individual target cell-killing events in vivo. Thus, the new mouse line is an ideal tool to study cytotoxic T lymphocyte biology and to optimize personalized immunotherapy in cancer treatment., Competing Interests: PC, HC, PM, CH, KR, EK, CS, TL, FB, JR No competing interests declared, MA works as a consultant for Biocrine, an unlisted biotech company that is using the ACE technique as a research tool. PB is cofounder and CEO of Biocrine, an unlisted biotech company that is using the ACE technique as a research tool. NB Reviewing editor, eLife, (© 2020, Chitirala et al.)

In a clinically-relevant model of 4 week, low-dose cisplatin-induced AKI, mice were injected subcutaneously with non small cell lung cancer (NSCLC) cells that harbor an activating Kirsten rat sarcoma viral oncogene homolog (KRAS) G12V mutation. Phospho extracellular signal-regulated kinase1/2 (pERK1/2) expression in kidney and tumors was decreased by the MEK1/2 inhibitors, U0126 and trametinib, that potently inhibit pERK1/2. U0126 resulted in a significant improvement in kidney function, acute tubular necrosis (ATN) and tubular cell apoptosis in mice with AKI. Genes that were significantly decreased by U0126 were heat shock protein 1, cyclin-dependent kinase 4 (CDK4) and stratifin (14-3-3σ). U0126 resulted in a significant decrease in tumor weight and volume and significantly increased the chemotherapeutic effect of cisplatin. Trametinib, a MEK1/2 inhibitor that is FDA-approved for the treatment of cancer, did not result in functional protection against AKI or worse AKI, but dramatically decreased tumor growth more than cisplatin. Smaller tumors in cisplatin or MEK1/2 inhibitor-treated mice were not related to changes in microtubule-associated proteins 1A/1B light chain 3B (LC3-II), p62, cleaved caspase-3, granzyme B, or programmed death-ligand 1 (PD-L1). In summary, despite ERK inhibition by both U0126 and trametinib, only U0126 protected against AKI suggesting that the protection against AKI by U0126 was due to an off-target effect independent of ERK inhibition. The effect of U0126 to decrease AKI may be mediated by inhibition of heat shock protein 1, CDK4 or stratifin (14-3-3σ). Trametinib was more effective than cisplatin in decreasing tumor growth, but unlike cisplatin, trametinib did not cause AKI., (Copyright © 2020 Elsevier Inc. All rights reserved.)

CTLs release cytotoxic proteins such as granzymes and perforin through fusion of cytotoxic granules (CG) at the target cell interface, the immune synapse, to kill virus-infected and tumorigenic target cells. A characteristic feature of these granules is their acidic pH inside the granule lumen, which is required to process precursors of granzymes and perforin to their mature form. However, the role of acidic pH in CG maturation, transport, and fusion is not understood. We demonstrate in primary murine CTLs that the a3-subunit of the vacuolar-type (H + )-adenosine triphosphatase is required for establishing a luminal pH of 6.1 inside CG using ClopHensorN(Q69M), a newly generated CG-specific pH indicator. Knockdown of the a3-subunit resulted in a significantly reduced killing of target cells and a >50% reduction in CG fusion in total internal reflection fluorescence microscopy, which was caused by a reduced number of CG at the immune synapse. Superresolution microscopy revealed a reduced interaction of CG with the microtubule network upon a3-subunit knockdown. Finally, we find by electron and structured illumination microscopy that knockdown of the a3-subunit altered the diameter and density of individual CG, whereas the number of CG per CTL was unaffected. We conclude that the a3-subunit of the vacuolar adenosine triphosphatase is not only responsible for the acidification of CG, but also contributes to the maturation and efficient transport of the CG to the immune synapse., (Copyright © 2020 by The American Association of Immunologists, Inc.)

This study focused on the synthesis, photocatalytic degradation of organic dyes and biological activity of novel N, N, O-donor tridentate water soluble 4-hydroxy benzohydrazide-grafted biopolymer Schiff base Cu(ii) complexes. The eco-friendly catalysts were designed for potential application in the degradation of organic dyes. The photocatalytic degradation of methylene blue was investigated with various irradiation times (30, 60, 90 and 120 min), catalytic dosages (5, 10, 15 and 20 mg) and pH (3, 7 and 12). The as-prepared compounds were characterised via various techniques including FT-IR and FT-NMR spectroscopy; TGA-DTA, XRD, SEM-EDAX; ESR and UV-vis spectroscopy; photoluminescence, magnetic moment, and conductivity measurements; and elemental and thermal analysis. The crystallinity of the Schiff base ligands, chitosan, and their Cu(ii) complexes was analysed via X-ray diffraction (XRD) studies. The XRD patterns revealed that the polymer chitosan was more crystalline than the Schiff base ligands and their complexes. The surface morphological analysis by scanning electron microscopy (SEM) revealed that the Cu(ii) complexes were amorphous in nature compared to chitosan and the ligands. The anti-inflammatory and anti-diabetic studies of the biopolymer Cu(ii) complexes were performed using the albumin denaturation technique and McCue and Shetty method, respectively. The as-synthesized 4-hydroxy benzohydrazide-grafted O -carboxymethyl chitosan Schiff base ligands and their Cu(ii) complexes showed a good anti-inflammatory and antidiabetic effect. The photocatalytic activity proved that the aryl-substituted complex was more efficient than the aliphatic-substituted complex., Competing Interests: There are no conflicts of interest, (This journal is © The Royal Society of Chemistry.)

A new series of lanthanum(iii) complexes was synthesized using a p -anisidine-appended 1-hydroxy-2-acetonapthanone (3) Schiff base and characterized via spectroscopic methods. The ligand was synthesized via sonication and the crystalline product was characterized using X-ray crystallography. The genotoxicity of the compound was assessed primarily by the bacterial reverse mutation (Ames) test and the in vitro mammalian chromosome aberration test; in both cases, the samarium complex 5 was found to be non-mutagenic. The anti-tumor activity of complexes 4, 5, and 6 was assayed against HeLa tumor cells and screened using the MTT assay. The IC 50 value of complex 5 was found to be 34 ± 1.2 μg mL -1 and this compound exhibited superior activity towards the cells compared to 4 and 6. These results were further confirmed by Hoechst 33258 staining and AO/EI dual staining, which indicated that the cells underwent an apoptosis mechanism in a dose-dependent manner. The apoptosis was further confirmed by the formation of ladders in the DNA fragmentation assay, and the western blot analysis of complex 5 suggested that the cells underwent the caspase-3-dependent pathway with PARP cleavage. Furthermore, the docking studies of complex 5 with HSA showed that it was situated in a hydrophilic cavity held by the electrostatic attraction of four hydrogen-bonding interactions. PDB ID:1BNA binds with complex 5 via strong π-π stacking interactions, which facilitate binding with the major grooves of DNA strands. The above-mentioned results illustrate that for complex 5, mitochondrion-mediated apoptosis occurs via caspase-3 activation. Complex 5 binds with DNA via intercalation because of S-phase cell cycle arrest in the HeLa cells., Competing Interests: All authors declare no conflict of interest., (This journal is © The Royal Society of Chemistry.)