Transcriptomic profiling of sex-specific olfactory neurons reveals subset-specific receptor expression in Caenorhabditis elegans.

The nematode Caenorhabditis elegans utilizes chemosensation to navigate an ever-changing environment for its survival. A class of secreted small-molecule pheromones, termed ascarosides, play an important role in olfactory perception by affecting biological functions ranging from development to behavior. The ascaroside #8 (ascr#8) mediates sex-specific behaviors, driving avoidance in hermaphrodites and attraction in males. Males sense ascr#8 via the ciliated male-specific cephalic sensory (CEM) neurons, which exhibit radial symmetry along dorsal-ventral and left-right axes. Calcium imaging studies suggest a complex neural coding mechanism that translates stochastic physiological responses in these neurons to reliable behavioral outputs. To test the hypothesis that neurophysiological complexity arises from differential expression of genes, we performed cell-specific transcriptomic profiling; this revealed between 18 and 62 genes with at least twofold higher expression in a specific CEM neuron subtype vs both other CEM neurons and adult males. These included two G protein-coupled receptor (GPCR) genes, srw-97 and dmsr-12, that were specifically expressed in nonoverlapping subsets of CEM neurons and whose expression was confirmed by GFP reporter analysis. Single CRISPR-Cas9 knockouts of either srw-97 or dmsr-12 resulted in partial defects, while a double knockout of both srw-97 and dmsr-12 completely abolished the attractive response to ascr#8. Together, our results suggest that the evolutionarily distinct GPCRs SRW-97 and DMSR-12 act nonredundantly in discrete olfactory neurons to facilitate male-specific sensation of ascr#8.

Ultrashort Peptide-Based Hydrogel for the Healing of Critical Bone Defects in Rabbits.

The use of hydrogels as scaffolds for three-dimensional (3D) cell growth is an active area of research in tissue engineering. Herein, we report the self-assembly of an ultrashort peptide, a tetrapeptide, Asp-Leu-IIe-IIe, the shortest peptide sequence from a highly fibrillogenic protein TDP-43, into the hydrogel. The hydrogel was mechanically strong and highly stable, with storage modulus values in MPa ranges. The hydrogel supported the proliferation and successful differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) in its matrix as assessed by cell viability, calcium deposition, alkaline phosphatase (ALP) activity, and the expression of osteogenic marker gene studies. To check whether the hydrogel supports 3D growth and regeneration in in vivo conditions, a rabbit critical bone defect model was used. Micro-computed tomography (CT) and X-ray analysis demonstrated the formation of mineralized neobone in the defect areas, with significantly higher bone mineralization and relative bone densities in animals treated with the peptide hydrogel compared to nontreated and matrigel treatment groups. The ultrashort peptide-based hydrogel developed in this work holds great potential for its further development as tissue regeneration and/or engineering scaffolds.

Quantitative evaluation of mercury adsorption and removal efficacy of Spirulina (Arthrospira platensis) powder in mice.

Spirulina is a blue-green alga, grown in alkaline water and used for detoxification of several toxic metal ions. Apart from its nutritional value, it is also used for the decontamination of toxic metal ions. Therefore, present study was envisaged to evaluate the adsorption and removal efficiency of Spirulina powder for mercury. The adsorption efficiency of Spirulina was evaluated in terms of weight of adsorbent, contact time, simulated gastric (SGF) and intestinal (SIF) fluid, and mercury concentration. In vivo removal efficacy of Spirulina for mercury was evaluated in mice. The mercury content in major tissues, urine and feces was estimated. The whole tissue retention and excretion of mercury after treatment with Spirulina were taken as a measure of its metal ions removal efficacy. Activated charcoal was taken as a standard adsorbent for comparative study. The maximum adsorption capacity of Spirulina and charcoal for mercury was found to be 66.667 and 158.730 mg g-1 in water, 83.33 and 94.340 mg g-1 in SGF and 125.0 and 133.33 mg g-1 in SIF, respectively. In mice, Spirulina and activated charcoal were significantly reduced the mercury deposition in tissues and facilitated their excretion through feces. Spirulina has shown good adsorption and removal efficacy like activated charcoal. Therefore, Spirulina can be used as a potential adsorbent to remove mercury from the body.

EEG based interpretation of human brain activity during yoga and meditation using machine learning: A systematic review.

OBJECTIVES: The present investigation is to study the impact of yoga and meditation on Brain waves concerning physical and mental health. There are mainly three stages (steps) in the brain wave classification:(i) preprocessing, ii) feature extraction, and iii) classification. This work provides a review of interpretation methods of Brain signals (Electroencephalogram (EEG)) EEG during yoga and meditation. Past research has revealed significant mental and physical advantages with yoga and meditation. METHODS: The research topic reviewed focused on the machine learning strategies applied for the interpretation of brain waves. In addressing the research questions highlighted earlier in the general introduction, we conducted a systematic search of articles from targeted scientific and journal online databases that included PubMed, Web of Science, IEEE Xplore Digital Library (IEEE), and Arxiv databases based on their relevance to the research questions and domain topic. The survey topic is relatively nascent, and therefore, the scope of the search period was limited to the 20-year timeline that was deemed representative of the research topic under investigation. The literature search was based on the keywords "EEG", "yoga*" and "meditation*". The key phrases were concatenated using Boolean expressions and applied to search through the selected online databases yielding a total of 120 articles. The online databases were selected based on the relevancy of content with the research title, research questions, and the domain application. The literature review search, process, and classification were carefully conducted guided by two defined measures; 1.) Inclusion criteria; and 2.) Exclusion criteria. These measures define the criteria for searching and extracting relevant articles relating to the research title and domain of interest. RESULTS: Our literature search and review indicate a broad spectrum of neural mechanics under a variety of meditation styles have been investigated. A detailed analysis of various mental states using Zen, CHAN, mindfulness, TM, Rajayoga, Kundalini, Yoga, and other meditation styles have been described by means of EEG bands. Classification of mental states using KNN, SVM, Random forest, Fuzzy logic, neural networks, Convolutional Neural Networks has been described. Superior research is still required to classify the EEG signatures corresponding to different mental states. CONCLUSIONS: Yoga practice may be an effective adjunctive treatment for a clinical and aging population. Advanced research can examine the effects of specific branches of yoga on a designated clinical grouping. Yoga and meditation increased overall healthy brain activity.

Impact of COVID-19 on Indian Pharmaceutical Industry and Way Forward.

BACKGROUND: As countries and industries continue to cope with the unparalleled challenges presented by the novel coronavirus (COVID-19), a specific area of concern has been the uncertainty surrounding the impact of the COVID-19 pandemic on the global and Indian supply chains of the pharmaceutical industry. The COVID-19 crisis has demonstrated the importance of establishing a risk management system that focuses on assessing future risks resulting from the loss of a supply chain among countries. OBJECTIVE: This review focuses on the role of the Indian pharmaceutical industry towards the pandemic. This review investigates the economic effect of COVID-19 across segments and what it implies for the Indian economy. METHODS: The COVID 19 flare-up has additionally commenced the Indian pharmaceutical organizations an opportunity to transform into a supported trade place point for gathering drugs and intermediates. RESULTS: An enormous pharmaceutical industry in India has consistently been a foundation of reasonable human services, and this pattern would now be able to be required to heighten further. CONCLUSION: The activities from COVID-19 are with a need to change the overall impression of Indian pharmaceutical associations and even more altogether, reduce the dependence of the private pharma associations on alone suppliers like China.

An Overview on Various Approaches and Recent Patents on Buccal Drug Delivery Systems.

BACKGROUND: Buccal delivery is an alluring course of organization for fundamental medication conveyance and it leads direct access to the systemic flow through the interior jugular vein sidesteps drugs from the hepatic first-pass digestion gives high bioavailability. OBJECTIVE: This article aims at buccal medication conveyance by discussing the structure and condition of the oral mucosa and the novel strategies utilized in evaluating buccal medication ingestion. METHODS: This review highlights the various pharmaceutical approaches for buccal delivery such as buccal tablets, buccal lozenges, buccal micro/nanoparticle, wafer and semisolid dosage forms like chewing gums, buccal patch, buccal gel or ointment and some buccal liquid dosage forms like buccal solutions and buccal sprays and recent patents filed or granted for these approaches. RESULTS: Recently, some patents are also reported where a combination of various approaches is being employed to achieve very effective mucosal delivery. The various patent search sites were used to collect and analyze the information on buccal drug delivery systems. CONCLUSION: The present study provides valuable information, advantages, limitations and future outlook of various buccal drug delivery systems.

Loss of Ceacam1 promotes prostate cancer progression in Pten haploinsufficient male mice.

OBJECTIVE: PTEN haploinsufficiency plays an important role in prostate cancer development in men. However, monoallelic deletion of Pten gene failed to induce high prostate intraepithelial neoplasia (PIN) until Pten+/- mice aged or fed a high-calorie diet. Because CEACAM1, a cell adhesion molecule with a potential tumor suppression activity, is induced in Pten+/- prostates, the study aimed at examining whether the rise of CEACAM1 limited neoplastic progression in Pten+/- prostates. METHODS: Pten+/- were crossbred with Cc1-/- mice harboring a null deletion of Ceacam1 gene to produce Pten+/-/Cc1-/- double mutants. Prostates from 7-month old male mice were analyzed histologically and biochemically for PIN progression. RESULTS: Deleting Ceacam1 in Pten+/- mice caused an early development of high-grade PIN in parallel to hyperactivation of PI3 kinase/Akt and Ras/MAP kinase pathways, with an increase in cell proliferation, epithelial-to-mesenchymal transition, angiogenesis and inflammation relative to Pten+/- and Cc1-/- individual mutants. It also caused a remarkable increase in lipogenesis in prostate despite maintaining insulin sensitivity. Concomitant Ceacam1 deletion with Pten+/- activated the IL-6/STAT3 signaling pathways to suppress Irf-8 transcription that in turn, led to a decrease in the expression level of promyelocytic leukemia gene, a well characterized tumor suppressor in prostate. CONCLUSIONS: Ceacam1 deletion accelerated high-grade prostate intraepithelial neoplasia in Pten haploinsufficient mice while preserving insulin sensitivity. This demonstrated that the combined loss of Ceacam1 and Pten advanced prostate cancer by increasing lipogenesis and modifying the STAT3-dependent inflammatory microenvironment of prostate.

Short to ultrashort peptide-based hydrogels as a platform for biomedical applications.

Short peptides have attracted significant attention from researchers in the past few years due to their easy design, synthesis and characterization, diverse functionalisation possibilities, low cost, possibility to make a large range of hierarchical nanostructures and most importantly their high biocompatibility and biodegradability. Generally, short peptides are also relatively more stable than their longer variants, non-immunogenic in nature and many of them self-assemble to provide an exciting range of nanostructures, including hydrogels. Thus, the development of short peptide-based hydrogels has become an area of intense investigation. Although these hydrogels have a water content of greater than 90%, they are surprisingly highly stable structures, and thus have been used for various biomedical applications, including cell therapeutics, drug delivery, tissue engineering and regeneration, contact lenses, biosensors, and wound healing, by different researchers. Herein, we review the progress of research in the rapidly expanding field of short to ultrashort peptide-based hydrogels and their possible applications. Special attention is paid to address and review this field with regard to the stability of peptide-based hydrogels, particularly to enzymatic degradation.

A polymorphism in intron I of the human angiotensinogen gene (hAGT) affects binding by HNF3 and hAGT expression and increases blood pressure in mice.

Angiotensinogen (AGT) is the precursor of one of the most potent vasoconstrictors, peptide angiotensin II. Genome-wide association studies have shown that two A/G polymorphisms (rs2493134 and rs2004776), located at +507 and +1164 in intron I of the human AGT (hAGT) gene, are associated with hypertension. Polymorphisms of the AGT gene result in two main haplotypes. Hap-I contains the variants -217A, -6A, +507G, and +1164A and is pro-hypertensive, whereas Hap-II contains the variants -217G, -6G, +507A, and +1164G and does not affect blood pressure. The nucleotide sequence of intron I of the hAGT gene containing the +1164A variant has a stronger homology with the hepatocyte nuclear factor 3 (HNF3)-binding site than +1164G. Here we found that an oligonucleotide containing +1164A binds HNF3ß more strongly than +1164G and that Hap-I-containing reporter gene constructs have increased basal and HNF3- and glucocorticoid-induced promoter activity in transiently transfected liver and kidney cells. Using a knock-in approach at the hypoxanthine-guanine phosphoribosyltransferase locus, we generated a transgenic mouse model containing the human renin (hREN) gene and either Hap-I or Hap-II. We show that transgenic animals containing Hap-I have increased blood pressure compared with those containing Hap-II. Moreover, the transcription factors glucocorticoid receptor, CCAAT enhancer-binding protein ß, and HNF3ß bound more strongly to chromatin obtained from the liver of transgenic animals containing Hap-I than to liver chromatin from Hap-II-containing animals. These findings suggest that, unlike Hap-II variants, Hap-I variants of the hAGT gene have increased transcription rates, resulting in elevated blood pressure.

Bioavailability Enhancement of Polymyxin B With Novel Drug Delivery: Development and Optimization Using Quality-by-Design Approach.

Polymyxin-B (Poly-B) is an effective antibiotic used to treat infections mainly caused due to sensitive gram-negative bacteria. They belong to the group of cyclic peptide antibiotics and are minimally absorbed from the gastrointestinal tract. This arises the need for bioavailability enhancement and is achieved in the present case using niosomes as carrier system. The Poly-B niosomes had been developed using Span 60 and cholesterol while optimization is achieved with quality-by-design (QBD) approach. In this QBD approach, 3 independent variables (Span 60:cholesterol, volume of phosphate-buffered saline [%], and amount of drug [mg]) each at 3 levels were studied. A total of 17 runs were suggested by the model which was further analyzed by optimizing 3 different responses (particle size, zeta potential, and entrapment efficiency [EE%]). The results had clearly shown that the optimum formulation selected by QBD was based on the criteria of attaining the maximum value of EE% and low value of size and zeta potential. Poly-B niosomes were further examined by in vitro antifungal, rat creatinine, and cytotoxicity assay. The pharmacokinetics and scintigraphy studies were also performed for in vivo behavior of Poly-B.

Tuning the structural and mechanical properties in ZrO2-SiO2 binary system through Y3+ inclusions.

The present investigation explores the influence of yttrium (Y3+) inclusions in ZrO2-SiO2 binary system (YSZ) on its structural and mechanical features. The powders were synthesized through sol-gel technique and an exclusive characterization were undertaken to investigate the structural and mechanical features influenced by Y3+ additions. Characterization techniques affirmed the crucial role of Y3+ on the resultant structural and thermal stability of the YZS system. Instrumented indentation inferred the enhanced mechanical properties demonstrated by YZS system in comparison with pure ZrO2-SiO2 binary system (ZS).

Design and fabrication of a magnetically actuated non-invasive reusable drug delivery device.

We present a novel approach of designing and fabricating a noninvasive drug delivery device which is capable of delivering the drug to the target site in a controlled manner. The device utilizes a reservoir which can be reused once the drug has completely diffused from it. This micro-reservoir based fabricated device has been successfully tested using niosomes of insulin drug filled in, which was then sealed with a magnetic membrane of 20 µm thick and was actuated by applying magnetic field. The deflection of the membrane on application of magnetic field results in the drug release from the reservoir. The discharge of the drug solution and the release rates was controlled by external magnetic field. The simulation of the membrane deflection using COMSOL software was carried out to optimize the concentration of the ferrous nanopowder in PDMS matrix. The characterization of the devices was implemented in-vitro on water and in-vivo on Wistar rats. It was also validated using high-performance liquid chromatography (HPLC) by observing characteristic peak of insulin. The blood samples showed the retention time of 2.79 min at λmax of 280 nm which further authenticated the effectiveness of the proposed work. This noninvasive fabricated device provides reusability, precise control and can enable the patient or a physician to actively administrate the drug when required.

Epicardial fat thickness: A surrogate marker of coronary artery disease - Assessment by echocardiography.

OBJECTIVE: Epicardial fat is considered as indicator of cardiovascular risk. Several studies have tested the association between epicardial fat thickness (EFT) and coronary artery disease. The aim of our study is to test the hypothesis that echocardiographic EFT is a marker of coronary artery disease. METHODS: One hundred and ten patients (70 males and 40 females with mean age of 51.5±10.6 and 52.6±9.6, respectively) admitted for coronary angiogram underwent assessment of epicardial fat thickness by echocardiography. Routine clinical examination, evaluation of risk factor profile, and anthropometric variables were also done. Epicardial fat thickness was measured on the free wall of right ventricle in parasternal long- and short-axis views at end-systole for 3 cardiac cycles. RESULTS: Mean epicardial fat thickness in angiographically normal patients and acute coronary syndromes were 4.4±1.2 and 6.9±1.9, respectively. Epicardial fat thickness in males and females were not statistically different. Burden of coronary arterial lesions denoted by Gensini score shows linear association with epicardial fat thickness and the severity of the coronary disease. CONCLUSION: Epicardial fat is independently and linearly associated with CAD and its severity.

Bilosomes: A Novel Approach to Meet the Challenges in Oral Immunization.

Oral immunization with vaccines has proven its worth by eradicating several diseases worldwide, yet only few oral vaccines exist in practice. This is because the oral route is plagued with various challenges for successful delivery of vaccines, such as limited absorption and high degradation in the gastrointestinal tract. Tremendous research in the last decade has made bilosomes a potential carrier system for oral immunization. Bilosomes with its name derived from bile salts (which is one of its major constituent), is a 'niosome-like' colloidal carrier. The collective information addressed through this review, convincingly points out that factors such as size, composition (type of bile salt, and lipid), charge, ligand attached, etc., which show significant influence on the type, and amount of immunity developed following the oral administration of bilosomes. These bile salts containing vesicular carriers have been employed to treat various serious conditions. Some of these methods have been successfully established into patents. Bilosomes loaded with antigens, peptides and other biological materials for the treatment of influenza, heart failure, diuresis, Irritable Bowel Syndrome (IBS) and other gastrointestinal disorders have been discussed in the present article.

Forced Hepatic Overexpression of CEACAM1 Curtails Diet-Induced Insulin Resistance.

Carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1) regulates insulin sensitivity by promoting hepatic insulin clearance. Liver-specific inactivation or global null-mutation of Ceacam1 impairs hepatic insulin extraction to cause chronic hyperinsulinemia, resulting in insulin resistance and visceral obesity. In this study we investigated whether diet-induced insulin resistance implicates changes in hepatic CEACAM1. We report that feeding C57/BL6J mice a high-fat diet reduced hepatic CEACAM1 levels by >50% beginning at 21 days, causing hyperinsulinemia, insulin resistance, and elevation in hepatic triacylglycerol content. Conversely, liver-specific inducible CEACAM1 expression prevented hyperinsulinemia and markedly limited insulin resistance and hepatic lipid accumulation that were induced by prolonged high-fat intake. This was partly mediated by increased hepatic ß-fatty acid oxidation and energy expenditure. The data demonstrate that the high-fat diet reduced hepatic CEACAM1 expression and that overexpressing CEACAM1 in liver curtailed diet-induced metabolic abnormalities by protecting hepatic insulin clearance.

High-calorie diet exacerbates prostate neoplasia in mice with haploinsufficiency of Pten tumor suppressor gene.

OBJECTIVE: Association between prostate cancer and obesity remains controversial. Allelic deletions of PTEN, a tumor suppressor gene, are common in prostate cancer in men. Monoallelic Pten deletion in mice causes low prostatic intraepithelial neoplasia (mPIN). This study tested the effect of a hypercaloric diet on prostate cancer in Pten (+/-) mice. METHODS: 1-month old mice were fed a high-calorie diet deriving 45% calories from fat for 3 and 6 months before prostate was analyzed histologically and biochemically for mPIN progression. Because Pten (+/-) mice are protected against diet-induced insulin resistance, we tested the role of insulin on cell growth in RWPE-1 normal human prostatic epithelial cells with siRNA knockdown of PTEN. RESULTS: In addition to activating PI3 kinase/Akt and Ras/MAPkinase pathways, high-calorie diet causes neoplastic progression, angiogenesis, inflammation and epithelial-mesenchymal transition. It also elevates the expression of fatty acid synthase (FAS), a lipogenic gene commonly elevated in progressive cancer. SiRNA-mediated downregulation of PTEN demonstrates increased cell growth and motility, and soft agar clonicity in addition to elevation in FAS in response to insulin in RWPE-1 normal human prostatic cells. Downregulating FAS in addition to PTEN, blunted the proliferative effect of insulin (and IL-6) in RWPE-1 cells. CONCLUSION: High-calorie diet promotes prostate cancer progression in the genetically susceptible Pten haploinsufficient mouse while preserving insulin sensitivity. This appears to be partly due to increased inflammatory response to high-caloric intake in addition to increased ability of insulin to promote lipogenesis.

Cell-specific proteomic analysis in Caenorhabditis elegans.

Proteomic analysis of rare cells in heterogeneous environments presents difficult challenges. Systematic methods are needed to enrich, identify, and quantify proteins expressed in specific cells in complex biological systems including multicellular plants and animals. Here, we have engineered a Caenorhabditis elegans phenylalanyl-tRNA synthetase capable of tagging proteins with the reactive noncanonical amino acid p-azido-L-phenylalanine. We achieved spatiotemporal selectivity in the labeling of C. elegans proteins by controlling expression of the mutant synthetase using cell-selective (body wall muscles, intestinal epithelial cells, neurons, and pharyngeal muscle) or state-selective (heat-shock) promoters in several transgenic lines. Tagged proteins are distinguished from the rest of the protein pool through bioorthogonal conjugation of the azide side chain to probes that permit visualization and isolation of labeled proteins. By coupling our methodology with stable-isotope labeling of amino acids in cell culture (SILAC), we successfully profiled proteins expressed in pharyngeal muscle cells, and in the process, identified proteins not previously known to be expressed in these cells. Our results show that tagging proteins with spatiotemporal selectivity can be achieved in C. elegans and illustrate a convenient and effective approach for unbiased discovery of proteins expressed in targeted subsets of cells.