Optimization of Newtonian fluid pressure in microcantilever integrated flexible microfluidic channel for healthcare application.

The demand for microfluidic pressure sensors is ever-increasing in various industries due to their crucial role in controlling fluid pressure within microchannels. While syringe pump setups have been traditionally used to regulate fluid pressure in microfluidic devices, they often result in larger setups that increase the cost of the device. To address this challenge and miniaturize the syringe pump setup, the researcher introduced integrated T-microcantilever-based microfluidic devices. In these devices, microcantilevers are incorporated, and their deflections correlate with the microchannel's pressure. When the relative pressure of fluid (plasma) changes, the T-microcantilever deflects, and the extent of this deflection provides information on fluid pressure within the microchannel. In this work, finite element method (FEM) based simulation was carried out to investigate the role of material, and geometric parameters of the cantilever, and the fluid viscosity on the pressure sensing capability of the T-microcantilever integrated microfluidic channel. The T-microcantilever achieves a maximum deflection of 127µm at a 5000µm/s velocity for Young's modulus(E) of 360 kPa of PDMS by employing a hinged structure. On the other hand, a minimum deflection of 4.05 × 10-5µm was attained at 5000µm/s for Young's modulus of 1 TPa for silicon. The maximum deflected angle of the T-cantilever is 20.46° for a 360 kPa Young's modulus while the minimum deflection angle of the T-cantilever is measured at 13.77° for 900 KPa at a fluid velocity of 5000µm s-1. The T-cantilever functions as a built-in microchannel that gauges the fluid pressure within the microchannel. The peak pressure, set at 8.86 Pa on the surface of the cantilever leads to a maximum deflection of 0.096µm (approximately 1µm) in the T-cantilever at a 1:1 velocity ratio. An optimized microfluidic device embedded with microchannels can optimize fluid pressure in a microchannel support cell separation.

Injection of human neuroblastoma cells into neural crest streams in live zebrafish embryos.

Cancer cell behavior is highly microenvironment dependent, but we have a limited understanding of malignant cell-microenvironment interactions in vivo. Here, we describe a protocol for xenotransplanting human neuroblastoma (NB) cells into streams of migrating neural crest stem cells in zebrafish embryos, followed by confocal time-lapse imaging and cell tracking. This high-resolution model system facilitates the quantitative spatiotemporal analysis of cancer cell-cell and cell-environment interactions. For complete details on the use and execution of this protocol, please refer to Treffy et al. (2021).

Machine Learning Technique to Detect and Classify Mental Illness on Social Media Using Lexicon-Based Recommender System.

The emergence of social media has allowed people to express their feelings on products, services, films, and so on. The feeling is the user's view or attitude towards any topic, object, event, or service. Overall, feelings have always influenced people's decision-making. In recent years, emotions have been analyzed intensively in natural language, but many problems still have to be watched. One of the most important problems is the lack of precise classification resources. Most of the research into feeling gradation is concerned with the issue of polarity grading, although, in many practical applications, this relatively grounded feeling measure is insufficient. Design methods are therefore essential, which can accurately classify feelings into a natural language. The principal goal of the research is to develop an overflow of grammatical rules-based classification of Indian language tweets. In this work, three main challenges are identified to classify feelings in Indian language tweets and possible methods for tackling such issues. Firstly, it has been found that the informal nature of tweets is crucial for the classification of feelings. Based on the tweets, the mental illness of the person has been classified. Therefore, to categorize Indian language tweets, a combination of grammar rules based on adjectives and negations is proposed. Secondly, people often express their feelings with slang words, abbreviations, and mixed words. A technique called field tags is used to include nongrammatical arguments such as slang words and diverse words. Thirdly, if a tweet is more complex, the morphological richness of the Indian language results in a loss of performance. The grammar rules are embedded in N-gram techniques and machine learning methods. These methods are grouped into three approaches, which functionally predict Indian language tweets with syntactic words.

Non-cardiac Manifestations in Adult Patients With Mucopolysaccharidosis.

Mucopolysaccharidoses (MPS) are a heterogeneous group of disorders that results in the absence or deficiency of lysosomal enzymes, leading to an inappropriate storage of glycosaminoglycans (GAGs) in various tissues of the body such as bones, cartilage, heart valves, arteries, upper airways, cornea, teeth, liver and nervous system. Clinical manifestations can become progressively exacerbated with age and affect their quality of life. Developments in advanced supportive treatment options such as enzyme replacement therapy (ERT), hematopoietic stem cell transplantation (HSCT) may have improved patients' life span. Adult MPS patients require specialist clinical surveillance long-term. In many cases, in addition to the MPS-related health problems, they may develop age-related complications. Considering the complexity of their clinical manifestations and lack of guidelines on the management of adult MPS disorders, multispecialty and multidisciplinary teams' care is essential to diagnose and treat health problems that are likely to be encountered. This review presents non-cardiac clinical manifestations, their pathophysiology, management and long-term outcomes in adult MPS patients.

A Liver Damage Prediction Using Partial Differential Segmentation with Improved Convolutional Neural Network.

Background: The liver is one of the most significant and most essential organs in the human body. It is divided into two granular lobes, one on the right and one on the left, connected by a bile duct. The liver is essential in the removal of waste products from human food consumption, the creation of bile, the regulation of metabolic activities, the cleaning of the blood by sensitizing digestive management, and the storage of vitamins and minerals. To perform the classification of liver illnesses using computed tomography (CT scans), two critical phases must first be completed: liver segmentation and categorization. The most difficult challenge in categorizing liver disease is distinguishing the liver from the other organs near it. Methodology. Liver biopsy is a kind of invasive diagnostic procedure, widely regarded as the gold standard for accurately estimating the severity of liver disease. Noninvasive approaches for examining liver illnesses, such as blood serum markers and medical imaging (ultrasound, magnetic resonance MR, and CT) have also been developed. This approach uses the Partial Differential Technique (PDT) to separate the liver from the other organs and Level Set Methodology (LSM) for separating the cancer location from the surrounding tissue based on the projected pictures used as input. With the help of an Improved Convolutional Classifier, the categorization of different phases may be accomplished. Results: Several accuracies, sensitivity, and specificity measurements are produced to assess the categorization of LSM using an Improved Convolutional classifier. Approximately, 97.5% of the performance accuracy of the liver categorization is achieved with a 94.5% continuous interval (CI) of [0.6775 1.0000] and an error rate of 2.1%. The suggested method's performance is compared to that of two existing algorithms, and the sensitivity and specificity provide an overall average of 96% and 93%, respectively, with 95% Continuous Interval of [0.7513 1.0000] and [0.7126 1.0000] for sensitivity and specificity, respectively.

Dietary lutein supplementation improves immunity and antioxidant status of captive Indian leopards (Panthera fusca).

Indian leopards kept in zoos are fed solely on carabeef on bone (CBB) diets. Carabeef contains lesser or no carotenoids. Hence, the captive Indian leopard diets are suspected to be deficient in carotenoids while their wild counterparts acquire these pigments from their natural prey. Lutein is a vital carotenoid that plays its role as an antioxidant and immunomodulator. This experiment investigates the effect of lutein supplementation on antioxidant status, immunity, and stress in captive Panthera fusca fed CBB diets. Nine leopards were used based on 3 × 3 replicated Latin square designs in the experiment. Groups CON, LUT20, and LUT40 were supplemented with 0, 20, and 40 ppm of lutein, respectively. Each experiment comprised of 10 days of wash-out period, 11 days of adaptation, and 4 days of collection. Digestibility of crude protein (CP) was higher (p < .01) in groups LUT20 and LUT40. Serum concentration of protein, globulin, urea (p < .05), total carotenoids, total antioxidant capacity (TAC), catalase (CAT) activity, and lymphocyte transformation test (LTT) index were higher (p < .001) in groups LUT20 and LUT40. Activity of superoxide dismutase (SOD) and serum concentration of immunoglobulin were higher (p < .001) in group LUT20. Serum concentration of malonaldehyde (MDA) and fecal concentration of cortisol decreased (p < .001) in groups LUT20 and LUT40. Serum concentration of total immunoglobulin (µg/ml) and LTT were higher in group LUT20. Fecal concentration of cortisol (ng/g) was lower in LUT20 and LUT40. The study concludes that supplementation of lutein at 20 ppm would improve antioxidant status and immunity and alleviate stress in captive Indian leopards.

Neuroblastoma differentiation in vivo excludes cranial tumors.

Neuroblastoma (NB), the most common cancer in the first year of life, presents almost exclusively in the trunk. To understand why an early-onset cancer would have such a specific localization, we xenotransplanted human NB cells into discrete neural crest (NC) streams in zebrafish embryos. Here, we demonstrate that human NB cells remain in an undifferentiated, tumorigenic state when comigrating posteriorly with NC cells but, upon comigration into the head, differentiate into neurons and exhibit decreased survival. Furthermore, we demonstrate that this in vivo differentiation requires retinoic acid and brain-derived neurotrophic factor signaling from the microenvironment, as well as cell-autonomous intersectin-1-dependent phosphoinositide 3-kinase-mediated signaling, likely via Akt kinase activation. Our findings suggest a microenvironment-driven explanation for NB's trunk-biased localization and highlight the potential for induced differentiation to promote NB resolution in vivo.

Health-seeking behavior and the initial economic impact of patients with open globe injuries seeking treatment in a tertiary care center in India.

PURPOSE: The aim of this study was to assess the health-seeking behavior and the costs of primary treatment in the immediate aftermath of open globe trauma among patients seeking care at a tertiary eye care center in South India. METHODS: This prospective observational study was carried out from June to December 2019. Details of the patient's demographic profile, health-seeking behavior immediately following the trauma and total cost incurred till the completion of primary treatment were collected. RESULTS: Eighty-five patients were recruited. Majority of patients were men (53, 82.8%), came from rural areas (65, 76.4%) and were the main breadwinners (44, 68%) of their family. After the initial trauma, a vast majority of the patients (68, 80%) visited the nearest eye care provider to obtain treatment and a majority of them (47, 69.1%) reached there within 3 h. The remaining patients (17, 20%) came directly to our center, the base hospital (BH). The mean distance travelled to the BH was 113.6 km. The mean total cost incurred was INR 20107.6 ± 10441.3. Approximately 84.8% of the patients reported a monthly income less than this amount. CONCLUSION: The economic impact of receiving primary definitive care following open globe injuries is higher than the average monthly income of more than 2/3rd of the patients. To replicate the success stories like the cataract outreach programs, there is a need to implement an integrated program encompassing workplace safety, legal protection to primary eye care providers and providing economic support for definitive treatment for the vulnerable population.

Notch signaling mediates olfactory multiciliated cell specification.

Epithelial multiciliated cells (MCCs) use motile cilia to direct external fluid flow, the disruption of which is associated with human diseases in a broad array of organs such as those in the respiratory, reproductive, and renal systems. While many of the signaling pathways that regulate MCC formation in these organ systems have been identified, similar characterization of MCC differentiation in the developing olfactory system has been lacking. Here, using live cell tracking, targeted cell ablation, and temporally-specific inhibition of the Notch signaling pathway, we identify the earliest time window of zebrafish olfactory MCC (OMCC) differentiation and demonstrate these cells' derivation from peridermal cells. We also describe regionally segregated Notch signaling across time points of rapid OMCC differentiation and show that Notch signaling downregulation yields an increase in OMCCs, suggesting that OMCC fate is normally repressed in a region-specific manner during olfactory development. Finally, we describe Notch signaling's regulation of the differentiation/ciliogenesis-associated genes foxj1a and foxj1b. Taken together, these findings provide new insights into the origins and developmental programming of OMCCs in vivo.

Addition of insoluble fiber to isolation media allows for increased metabolite diversity of lab-cultivable microbes derived from zebrafish gut samples.

There is a gap in measured microbial diversity when comparing genomic sequencing techniques versus cultivation from environmental samples in a laboratory setting. Standardized methods in artificial environments may not recapitulate the environmental conditions that native microbes require for optimal growth. For example, the intestinal tract houses microbes at various pH values as well as minimal oxygen and light environments. These microbes are also exposed to an atypical source of carbon: dietary fiber compacted in fecal matter. To investigate how the addition of insoluble fiber to isolation media could affect the cultivation of microbes from zebrafish intestines, an isolate library was built and analyzed using the bioinformatics pipeline IDBac. While all isolation media encouraged the growth of species from several phyla, the extent of growth was greater with the addition of fiber allowing for easier isolation. Furthermore, fiber addition altered the metabolism of the cultivated gut-derived microbes and induced the production of unique metabolites that were not produced when microbes were otherwise grown on standard isolation media. Addition of this inexpensive carbon source to the media supported the cultivation of a diverse community whose secondary metabolite production may more closely replicate their metabolite production in vivo.

Tracking neural crest cell cycle progression in vivo.

Analysis of cell cycle entry/exit and progression can provide fundamental insights into stem cell propagation, maintenance, and differentiation. The neural crest is a unique stem cell population in vertebrate embryos that undergoes long-distance collective migration and differentiation into a wide variety of derivatives. Using traditional techniques such as immunohistochemistry to track cell cycle changes in such a dynamic population is challenging, as static time points provide an incomplete spatiotemporal picture. In contrast, the fluorescent, ubiquitination-based cell cycle indicator (Fucci) system provides in vivo readouts of cell cycle progression and has been previously adapted for use in zebrafish. The most commonly used Fucci systems are ubiquitously expressed, making tracking of a specific cell population challenging. Therefore, we generated a transgenic zebrafish line, Tg(-4.9sox10:mAG-gmnn(1/100)-2A-mCherry-cdt1(1/190)), in which the Fucci system is specifically expressed in delaminating and migrating neural crest cells. Here, we demonstrate validation of this new tool and its use in live high-resolution tracking of cell cycle progression in the neural crest and derivative populations.

Interferon induced Mx protein from Indian snow trout Schizothorax richardsonii (Gray) lacks critical functional features unlike its mammalian homologues.

Viral attack within host cells triggers the production of type I interferons and leads to the induction of interferon stimulated genes (ISGs). One of the ISG Mx, encodes type I interferon inducible GTPase that is responsible for the establishment of an anti-viral state within cells. Intriguingly, several isoforms of Mx have been reported in fish, but the structural analysis of fish Mx proteins remains unexplored. For the first time, we have identified and unraveled the molecular structure of Mx protein from Indian snow trout, Schizothorax richardsonii (Gray) a Coldwater fish that inhabits the water bodies in the sub-Himalayan region. The snow trout Mx coding region consists of 2518 nucleotides with an open reading frame (ORF) of 1854 nucleotides. It codes for a polypeptide of 617 amino acids with a predicted molecular weight of 70 kDa. In silico analysis of snow trout Mx protein revealed signature of dynamin family (LPRGTGIVTR) along with a tripartite GTP-binding domain (GDQSSGKS, DLPG, and TKPD). Homology modelling established that the Mx protein is an elongated structure with a G domain, bundle signaling element (BSE) and a GTPase effector domain (GED). Moreover, the GED of Mx contains two highly conserved leucine zippers at the COOH-terminal of the protein suggesting its structural similarity with human homologues. However, snow trout Mx lacks the essential features of its mammalian homologues questioning its functional characteristics. Further, a ligand binding site in the said protein has also been predicted adjacent to the GTPase switch within the G domain.

Spontaneous regression of retro-odontoid post traumatic cicatrix following occipitocervical fixation.

This case report describes a patient who presented with myelopathy secondary to a large retro-odontoid post traumatic cicatrix. The objective of this study was to discuss the clinical presentation, pathogenesis, imaging, and surgical management of pseudoarthrosis tissue mass associated with odontoid nonunion. Atlantoaxial subluxation (AAS) has been widely reported in patients with rheumatoid arthritis. AAS leads to repeated cycles of partial tear and repair of ligaments around the altantoaxial complex, resulting in the formation of periodontoid mass (pseudotumor). It is thought that formation of retro-odontoid post traumatic mass (cicatrix), in certain cases of odontoid fracture, is because of similar pathology. This is a retrospective review of case note. Here, the patient underwent posterior decompression through a C1-C2 laminectomy and occipitocervical (C0-C4) fusion with instrumentation, which resulted in dramatic improvement in his symptoms and spontaneous regression of retro-odontoid post traumatic cicatrix. We have described an interesting and a rare case of a large pseudoarthrosis tissue mass associated with odontoid nonunion, which regressed following stand-alone posterior instrumentation. To the best of our knowledge, only a handful of such cases of spontaneous regression of retro-odontoid post traumatic cicatrix following occipitocervical fixation have been described in literature, and our case adds to the growing list of such cases and may help in understanding the natural history of the disease process one day. Although rare, post traumatic cicatrix should be considered as a differential diagnosis of enhancing retro-odontoid mass, especially if there is any history of cervical spine trauma.

Targeted Pth4-expressing cell ablation impairs skeletal mineralization in zebrafish.

Skeletal development and mineralization are essential processes driven by the coordinated action of neural signals, circulating molecules and local factors. Our previous studies revealed that the novel neuropeptide Pth4, synthesized by hypothalamic cells, was involved in bone metabolism via phosphate regulation in adult zebrafish. Here, we investigate the role of pth4 during skeletal development using single-cell resolution, two-photon laser ablation of Pth4:eGFP-expressing cells and confocal imaging in vivo. Using a stable transgenic Pth4:eGFP zebrafish line, we identify Pth4:eGFP-expressing cells as post-mitotic neurons. After targeted ablation of eGFP-expressing cells in the hypothalamus, the experimental larvae exhibited impaired mineralization of the craniofacial bones whereas cartilage development was normal. In addition to a decrease in pth4 transcript levels, we noted altered expression of phex and entpd5, genes associated with phosphate homeostasis and mineralization, as well as a delay in the expression of osteoblast differentiation markers such as sp7 and sparc. Taken together, these results suggest that Pth4-expressing hypothalamic neurons participate in the regulation of bone metabolism, possibly through regulating phosphate balance during zebrafish development.

Neural crest and cancer: Divergent travelers on similar paths.

Neural crest cells are multipotent progenitors that dynamically interpret diverse microenvironments to migrate significant distances as a loosely associated collective and contribute to many tissues in the developing vertebrate embryo. Uncovering details of neural crest migration has helped to inform a general understanding of collective cell migration, including that which occurs during cancer metastasis. Here, we discuss several commonalities and differences of neural crest and cancer cell migration and behavior. First, we focus on some of the molecular pathways required for the initial specification and potency of neural crest cells and the roles of many of these pathways in cancer progression. We also describe epithelial-to-mesenchymal transition, which plays a critical role in initiating both neural crest migration and cancer metastasis. Finally, we evaluate studies that demonstrate myriad forms of cell-cell and cell-environment communication during neural crest and cancer collective migration to highlight the remarkable similarities in their molecular and cell biological regulation.

Cell and small animal models for phenotypic drug discovery.

The phenotype-based drug discovery (PDD) approach is re-emerging as an alternative platform for drug discovery. This review provides an overview of the various model systems and technical advances in imaging and image analyses that strengthen the PDD platform. In PDD screens, compounds of therapeutic value are identified based on the phenotypic perturbations produced irrespective of target(s) or mechanism of action. In this article, examples of phenotypic changes that can be detected and quantified with relative ease in a cell-based setup are discussed. In addition, a higher order of PDD screening setup using small animal models is also explored. As PDD screens integrate physiology and multiple signaling mechanisms during the screening process, the identified hits have higher biomedical applicability. Taken together, this review highlights the advantages gained by adopting a PDD approach in drug discovery. Such a PDD platform can complement target-based systems that are currently in practice to accelerate drug discovery.

A Confirmed Case in the United Kingdom of Hirayama Disease in a Young White Male Presenting with Hand Weakness.

BACKGROUND: We present the first reported case of Hirayama disease in the United Kingdom. A literature review of Hirayama disease in the Western literature shows that this case is unique in being the first in the United Kingdom, constituting one of only a few cases in Europe with supporting magnetic resonance imaging and reported treatment outcome. CASE DESCRIPTION: Our patient was a young Caucasian male who presented with progressive bilateral hand weakness, had confirmatory magnetic resonance imaging findings of Hirayama disease, and experienced improvement of symptoms with cervical collar immobilization. CONCLUSIONS: This case lends further evidence to the flexion-induced myelopathy theory of Hirayama disease and supports the findings of other studies in which avoidance of neck flexion and cervical immobilization helped to attenuate disease progression in this patient group.

Alum adjuvanted rabies DNA vaccine confers 80% protection against lethal 50 LD50 rabies challenge virus standard strain.

Rabies is a serious concern world-wide. Despite availability of rabies vaccines for long; their efficacy, safety, availability and cost effectiveness has been a tremendous issue. This calls for improvement of rabies vaccination strategies. DNA vaccination has immense potential in this regard. The DNA vaccine pgp.LAMP-1 conferred 60% protection to BALB/c mice against 20 LD50 rabies challenge virus standard (CVS) strain challenge. Upon supplementation with Emulsigen-D, the vaccine formulation conferred complete protection against lethal challenge. To assess the feasibility of this vaccine formulation for human use, it was tested along with other FDA approved adjuvants, namely, Alum, Immuvac, Montanide ISA720 VG. Enhanced immune response correlated with high IgG antibody titer, Th2 biased response with a high level of rabies virus neutralizing antibodies (RVNAs) and IgG1/IgG2a ratio >1, observed upon alum supplementation of the rabies DNA vaccine. The total IgG antibody titer was 2IU/ml and total RVNA titer was observed to be 4IU/ml which is eight times higher than the minimum protective titer recommended by WHO. Furthermore, it conferred 80% protection against challenge with 50 LD50 of the rabies CVS strain, conducted in compliance with the potency test for rabies recommended by the National Institutes of Health (NIH), USA. Previously, we have established pre-clinical safety of this vaccine as per the guidelines of Schedule Y, FDA as well as The European Agency for evaluation of Medicinal Products. The vaccine showed no observable toxicity at the site of injection as well as at systemic level in Wistar rats when administered with 10X recommended dose. Therefore, supplementation of rabies DNA vaccine, pgp.LAMP-1 with alum would lead to development of a non-toxic, efficacious, stable and affordable vaccine that can be used to combat high numbers of fatal rabies infections tormenting developing countries.