Three million images and morphological profiles of cells treated with matched chemical and genetic perturbations.

The identification of genetic and chemical perturbations with similar impacts on cell morphology can elucidate compounds' mechanisms of action or novel regulators of genetic pathways. Research on methods for identifying such similarities has lagged due to a lack of carefully designed and well-annotated image sets of cells treated with chemical and genetic perturbations. Here we create such a Resource dataset, CPJUMP1, in which each perturbed gene's product is a known target of at least two chemical compounds in the dataset. We systematically explore the directionality of correlations among perturbations that target the same protein encoded by a given gene, and we find that identifying matches between chemical and genetic perturbations is a challenging task. Our dataset and baseline analyses provide a benchmark for evaluating methods that measure perturbation similarities and impact, and more generally, learn effective representations of cellular state from microscopy images. Such advancements would accelerate the applications of image-based profiling of cellular states, such as uncovering drug mode of action or probing functional genomics.

Discovery of reversible and covalent TEAD 1 selective inhibitors MSC-1254 and MSC-5046 based on one scaffold.

The Transcriptional Enhanced Associated Domain (TEAD) family of transcription factors are key components of the Hippo signalling family which play a crucial role in the regulation of cell proliferation, differentiation and apoptosis. The identification of inhibitors of the TEAD transcription factors are an attractive strategy for the development of novel anticancer therapies. A HTS campaign identified hit 1, which was optimised using structure-based drug design, to deliver potent TEAD1 selective inhibitors with both a reversible and covalent mode of inhibition. The preference for TEAD1 could be rationalised by steric differences observed in the lower pocket of the palmitoylation-site between subtypes, with TEAD1 having the largest available volume to accommodate substitution in this region.

The Cardiac Comeback-Beating Stronger: Exploring the Remarkable Resilience of the Heart in COVID-19 Recovery through Cardiac Autonomic Analysis.

Background and Objectives: Analyzing the cardiac autonomic function in COVID-19 patients can provide insights into the impact of the virus on the heart's regulatory mechanisms and its recovery. The autonomic nervous system plays a crucial role in regulating the heart's functions, such as heart rate, blood pressure, and cardiac output. This study aimed to investigate the impact of COVID-19 on heart rate variability (HRV) during a 6-min walk test (6MWT). Materials and Methods: The study included 74 participants, consisting of 37 individuals who had recovered from mild to moderate COVID-19 and 37 healthy controls. The study assessed heart rate variability (HRV) and blood pressure both before and after a 6-min walk test (6MWT). Results: The study found significant differences in a few time domains (SDNN and pNN50) and all frequency domain measures, whereas there were no significant differences in demographic characteristics or blood pressure between COVID-19-recovered individuals and healthy controls at rest. There were significant 6MWT effects on average HR, time-domain (SDNN and pNN50) measures of HRV, and all frequency domain measures of HRV. A significant group × 6MWT interaction was found for SDNN, pNN50, total power, Ln total power, LF, HF, Ln LF, Ln HF, and LF nu. Conclusions: Cardiac Autonomic analysis through HRV is essential to ensure the continued health and well-being of COVID-19 survivors and to minimize the potential long-term impacts of the disease on their cardiovascular system. This suggests that HRV analysis during the recovery phase following exercise could serve as a valuable tool for evaluating the physiological effects of COVID-19 and monitoring the recovery process.

A Microfluidic 3D Endothelium-on-a-Chip Model to Study Transendothelial Migration of T Cells in Health and Disease.

The recruitment of T cells is a crucial component in the inflammatory cascade of the body. The process involves the transport of T cells through the vascular system and their stable arrest to vessel walls at the site of inflammation, followed by extravasation and subsequent infiltration into tissue. Here, we describe an assay to study 3D T cell dynamics under flow in real time using a high-throughput, artificial membrane-free microfluidic platform that allows unimpeded extravasation of T cells. We show that primary human T cells adhere to endothelial vessel walls upon perfusion of microvessels and can be stimulated to undergo transendothelial migration (TEM) by TNFα-mediated vascular inflammation and the presence of CXCL12 gradients or ECM-embedded melanoma cells. Notably, migratory behavior was found to differ depending on T cell activation states. The assay is unique in its comprehensiveness for modelling T cell trafficking, arrest, extravasation and migration, all in one system, combined with its throughput, quality of imaging and ease of use. We envision routine use of this assay to study immunological processes and expect it to spur research in the fields of immunological disorders, immuno-oncology and the development of novel immunotherapeutics.

Small molecules-Giant leaps for immuno-oncology.

Immuno-oncology therapies are revolutionizing the oncology landscape with checkpoint blockade becoming the treatment backbone for many indications. While inspiring, much work remains to increase the number of cancer patients that can benefit from these treatments. Thus, a new era of immuno-oncology research has begun which is focused on identifying novel combination regimes that lead to improved response rates. This review highlights the significance of small molecules in this approach and illustrates the huge progress that has been made to date.

Comparison of Dimensional Accuracy of Stone Models Fabricated by Three Different Impression Techniques Using Two Brands of Polyvinyl Siloxane Impression Materials.

AIM: The aim of this study is to check the dimensional precision of stone models made by two different brands of polyvinyl siloxane impression materials using the monophase, one-step, and two-step putty/light-body impression techniques. MATERIALS AND METHODS: A metal model, having two crown preparations, was fabricated. With each technique, 40 impressions were made using two types of polyvinyl siloxane impression materials (Aquasil and Virtual). A total of 240 impressions were made with both the polyvinyl siloxane impression materials. Monophase impressions were made with a medium body using an acrylic custom tray. By simultaneous usage of putty and light-body, one-step impressions were made with a perforated metal stock tray. For two-step impressions, a 25-40 microns thick cellophane sheet spacer was used. The stone casts were obtained from the impressions of the stainless steel model. Three different dimensions (height, diameter, and inter-abutment distance) on these resultant stone casts were compared with the standard die. The accuracy of two different brands of impression materials was also compared. The results were then statistically analyzed. RESULTS: The resultant casts obtained from the different impression techniques had significantly larger dimensions in height and diameter, but smaller dimensions were observed for the inter-abutment distance. Larger deviation in resultant casts was observed in the monophase than one-step impression technique and the least deviation was observed in the two-step impression technique. CONCLUSION: The two-step impression technique produced the most accurate results in terms of the resultant casts. Out of the two different brands, Aquasil produced more fare results. CLINICAL SIGNIFICANCE: Adequate marginal adaptation, proper fit and least distortion of the castings, and the final prosthesis can be achieved by using the adequate impression technique and impression material.

Direct visualization of newly synthesized target proteins in situ.

Protein synthesis is a dynamic process that tunes the cellular proteome in response to internal and external demands. Metabolic labeling approaches identify the general proteomic response but cannot visualize specific newly synthesized proteins within cells. Here we describe a technique that couples noncanonical amino acid tagging or puromycylation with the proximity ligation assay to visualize specific newly synthesized proteins and monitor their origin, redistribution and turnover in situ.

Medical students' attitudes towards well-being and welfare: a systematic review protocol.

INTRODUCTION: Medical education poses serious stressors on medical students, as they report overall higher rates of psychiatric comorbidities compared with students enrolled in other university-level courses. The high rates of poor well-being reported should be considered a concern, as students represent the future workforce of healthcare professionals. Although there has been an increased interest towards well-being and burn-out in medical students, there is still much to be elucidated. Indeed, there is an overall lack of understanding of which attitudes medical students have towards well-being and welfare, something that could be used to develop targeted practice to improve medics' quality of life and reduce overall burn-out.This review will aim at evaluating and reporting the current research available on medical students' attitudes towards well-being and welfare. We will take into account which countries have published data on the topic, and we will analyse papers written during the COVID-19 period separately, giving this particular time frame a separate outlook. METHODS AND ANALYSIS: This systematic review protocol has been developed following Preferred Reporting Items for Systematic Review and Meta-Analysis Protocols guidelines. A systematic search including "medical students" AND "wellbeing" AND "perceptions" was carried out on MEDLINE, Embase and Scopus databases. The Mixed Method Appraisal Tool will be used to assess the quality of the available studies and risk of bias. Studies will be screened using predetermined inclusion and exclusion criteria. Data will be summarised narratively and in tabular forms. A narrative subgroup analysis of the COVID-19 period will be carried out, and a country-wise approach will be used to describe which areas have published relevant studies. ETHICS AND DISSEMINATION: This will be the first systematic review looking at the reported attitudes that medical students have towards well-being and welfare. This will provide an account of the available data on the topic, and a starting point to understand where further research is needed in the future. PROSPERO REGISTRATION NUMBER: CRD42023471022.

Understanding the emergence of negative photoconductivity in CVD grown ReS2 thin films by invoking the trion generation mechanism.

Emerging from robust coulombic interactions and diminished dielectric screening, two-dimensional transition metal dichalcogenides (2D TMDs) manifest strongly bound excitons. While these pronounced many-body effects have been observed in 2D TMDs, their impact on the materials' intrinsic photoconductive characteristics remains relatively less explored. In this study, a considerable reduction in photoconductivity in chemical vapor deposited (CVD) ReS2 thin films has been observed. Through experimental and theoretical scrutiny, it is deduced that this anomalous phenomenon stems from the possible many-body interactions within the system, wherein photoexcited electron-hole pairs combine with excess electrons to generate trions. The formation of trions reduces the effective number of carriers contributing to photocurrent, thereby attenuating the photoconductivity. This study shows the profound influence of trions on the photoconductivity behaviour of ReS2 thin films, thereby elucidating their significance in modulating optical and electrical properties crucial for optoelectronic applications.

Cystic presentation of primary hepatic neuroendocrine tumour: a case report with a brief review of literature.

Neuroendocrine tumours (NETs) are a rare type of tumours that arise from the neuroendocrine cells which are distributed throughout the body. Of all the gastrointestinal tumours only 1-2% account for NETs. They have an extremely low incidence of 0.17% arising in the intrahepatic bile duct epithelium. Majority of hepatic NETs occur as a result of metastases from the primary NETs. Most cases of primary hepatic NET (PHNET) present as a solid nodular mass. However, predominantly cystic PHNET is extremely rare which mimics other cystic space-occupying lesions clinically and radiologically as seen in this case.

Recent Progress and Challenges in the Development of Edible Vaccines Produced by Genetically Modified Plants.

Biotechnologists have pioneered the idea of an edible vaccination in recent years. Subunit vaccines, such as those used to create edible vaccines, involve the introduction of certain genes into transgenic plants, which are subsequently coaxed into producing the corresponding protein. Bananas, potatoes, legumes, lettuce, soybeans, corn, and rice are all examples of foods that fall under this category. They have a low unit cost, can be stored conveniently, and are simple to administer to patients of varying ages. There is great hope that the use of edible vaccinations, particularly in underdeveloped countries, could drastically reduce the prevalence of diseases, including measles, cholera, hepatitis B, and diarrhea. The development of effective and widely applicable edible vaccination, however, faces a number of technological and regulatory hurdles. When compared to traditional immunizations, edible vaccines offer significant cost savings, increased productivity, and reduced risk. It raises the possibility of a more efficient approach to illness prevention. This article includes important uses, production, host plants, benefits, drawbacks, mechanism of action, and many regulatory difficulties related to edible vaccines. In this article, we have discussed the most recent developments and successes with edible and intradermal vaccines in terms of the system used for immunogen production, the molecular properties of these vaccines, and their ability to generate a protective systemic and mucosal response.

Functional and Tableting Properties of Alkali-Isolated and Phosphorylated Barnyard Millet (Echinochloa esculenta) Starch.

The functional and tableting properties of barnyard millet starch (Echinochloa esculenta) were investigated in its native (alkali-treated) and chemically modified (phosphorylated) states. The grains were pulverized, soaked, and ground before filtration to separate starch and protein. Multiple NaOH treatments were performed. The starch was washed, neutralized, and dried. Sodium tripolyphosphate (STPP) and sodium sulfate were used to modify the starch, followed by maceration, washing, and drying to remove unreacted chemicals. The amylose content of alkali-treated barnyard millet starch increased by 19.96 ± 3.56% w/w. The amount of protein, the kind of starch used, and the size of the starch granules, all affected the ability of the starch granules to swell up. It was observed that alkali-extracted barnyard millet starch (AZS) has a swelling power of 194.3 ± 0.0064% w/w. The swelling capacity of treated starch was lesser as compared to the native alkali barnyard millet starch. Decrement in swelling power of phosphorylated starch was observed due to tightening of bonds in the molecular structure. The moisture content of the excipients may affect the overall stability of the formulation. The moisture content of the AZS was found to be 15.336 ± 1.012% w/w. Compared to AZS, cross-linked barnyard millet starch had a moisture content that was up to 20% lower than AZS. The Hausner ratio for phosphorylated starch was found to be 1.25, which indicates marked flow property. Similar morphologies could be seen in the alkali-isolated barnyard millet starch and the cross-linked/phosphorylated barnyard millet that was cross-linked using a mixture of sodium sulfate and sodium tripolyphosphate. The modest degree of substitution would have no effect on the surface morphology as shown by the scanning electron microscopic study. The crushing and compacting abilities of modified barnyard millet starch were also improved, but its friability and rate of disintegration were decreased. The whole study revealed that after cross-linking, barnyard millet had good tableting properties and it can be used as an excipient in drug delivery.

Optimizing the Cell Painting assay for image-based profiling.

In image-based profiling, software extracts thousands of morphological features of cells from multi-channel fluorescence microscopy images, yielding single-cell profiles that can be used for basic research and drug discovery. Powerful applications have been proven, including clustering chemical and genetic perturbations on the basis of their similar morphological impact, identifying disease phenotypes by observing differences in profiles between healthy and diseased cells and predicting assay outcomes by using machine learning, among many others. Here, we provide an updated protocol for the most popular assay for image-based profiling, Cell Painting. Introduced in 2013, it uses six stains imaged in five channels and labels eight diverse components of the cell: DNA, cytoplasmic RNA, nucleoli, actin, Golgi apparatus, plasma membrane, endoplasmic reticulum and mitochondria. The original protocol was updated in 2016 on the basis of several years' experience running it at two sites, after optimizing it by visual stain quality. Here, we describe the work of the Joint Undertaking for Morphological Profiling Cell Painting Consortium, to improve upon the assay via quantitative optimization by measuring the assay's ability to detect morphological phenotypes and group similar perturbations together. The assay gives very robust outputs despite various changes to the protocol, and two vendors' dyes work equivalently well. We present Cell Painting version 3, in which some steps are simplified and several stain concentrations can be reduced, saving costs. Cell culture and image acquisition take 1-2 weeks for typically sized batches of ≤20 plates; feature extraction and data analysis take an additional 1-2 weeks.This protocol is an update to Nat. Protoc. 11, 1757-1774 (2016): https://doi.org/10.1038/nprot.2016.105.

Air flow limitation in smokers - A cause of concern.

Introduction: In India smoking is a common habit prevalent in both urban and rural areas irrespective of mode of smoking i.e., cigarettes, bidis, pipes, cigar, hookah etc., Spirometry can be helpful to determine effects of smoking on pulmonary functions. We aimed to study the effect of smoking on the pulmonary function tests. Materials and Methods: This study was conducted on 300 subjects including 150 smokers and 150 non-smokers aged between 25 and 60 years attending a tertiary health care center in northern part of our country. Quantification of tobacco smoking was performed by calculating smoking index. All the study subjects underwent spirometry. Results: All the spirometric variables (FVC, FEV1, PEFR, FEF 25-75%) were lower in smokers as compared to non-smokers and this difference was statistically significant. 76% of the smokers had obstructive, 10.7% had normal, 6.7% had restrictive, and 6.7% had mixed pattern on spirometry. 65.3% of the non-smokers had normal, 28.7% had obstructive and 6% had restrictive pattern on spirometry. Conclusion: Almost all the pulmonary function parameters were significantly reduced in smokers compared to non-smokers and obstructive impairment was common amongst smokers. As early quitting is associated with improved survival, it is important that these asymptomatic smokers are identified early and helped to quit. Primary care physicians being the first point of contact, can play a major role.

Optimization of TEAD P-Site Binding Fragment Hit into In Vivo Active Lead MSC-4106.

The dysregulated Hippo pathway and, consequently, hyperactivity of the transcriptional YAP/TAZ-TEAD complexes is associated with diseases such as cancer. Prevention of YAP/TAZ-TEAD triggered gene transcription is an attractive strategy for therapeutic intervention. The deeply buried and conserved lipidation pocket (P-site) of the TEAD transcription factors is druggable. The discovery and optimization of a P-site binding fragment (1) are described. Utilizing structure-based design, enhancement in target potency was engineered into the hit, capitalizing on the established X-ray structure of TEAD1. The efforts culminated in the optimized in vivo tool MSC-4106, which exhibited desirable potency, mouse pharmacokinetic properties, and in vivo efficacy. In close correlation to compound exposure, the time- and dose-dependent downregulation of a proximal biomarker could be shown.

Large falciform ligament cyst in a child: A rare entity of peritoneal cysts and review of the literature.

Peritoneal cysts are not uncommon in children - mesenteric/omental cysts being the commoner entity. Peritoneal cysts in the falciform ligament are a very rare entity reported in the literature. We present a 5-year-old boy who presented with pain upper central abdomen and few episodes of non-bilious vomiting for 1 year. He was stable on examination, with abdominal examination revealing the fullness of the abdomen with palpable generalised cystic mass which was mobile transversely. Ultrasound and contrast-enhanced computed tomography of the abdomen revealed intra-peritoneal cystic lesion measuring 13 cm × 11.5 cm × 9 cm with septations seen in the abdominal cavity from the epigastric region to the infraumbilical region (D11-L5 level). Laparoscopy showed a large cyst of the abdomen arising from the anterior abdominal wall, along the falciform ligament. The dark yellow fluid was aspirated and the cyst was excised leaving a part which was adherent to the anterior wall. Histopathology revealed cuboidal epithelium lined by the fibrous wall. The patient is doing well at 6 months follow-up. Falciform ligament cysts are very rare and laparoscopy can confirm the diagnosis as well as help in excision of the cyst with good results.

Evaluation of Remote Monitoring Device for Monitoring Vital Parameters against Reference Standard: A Diagnostic Validation Study for COVID-19 Preparedness.

CONTEXT: Vital parameters including blood oxygen level, respiratory rate, pulse rate, and body temperature are crucial for triaging patients to appropriate medical care. Advances in remote health monitoring system and wearable health devices have created a new horizon for delivery of efficient health care from a distance. MATERIALS AND METHODS: This diagnostic validation study included patients attending the outpatient department of the institute. The accuracy of device under study was compared against the gold standard patient monitoring systems used in intensive care units. STATISTICAL ANALYSIS: The statistical analysis involved computation of intraclass correlation coefficient. Bland-Altman graphs with limits of agreement were plotted to assess agreement between methods. P <0.05 was considered statistically significant. RESULTS: A total of 200 patients, including 152 males and 48 females in the age range of 2-80 years, formed the study group. A strong correlation (intraclass correlation coefficient; r > 0.9) was noted between the two devices for all the investigated parameters with significant P value (<0.01). Bland-Altman plot drawn for each vital parameter revealed observations in agreement from both the devices. CONCLUSION: The wearable device can be reliably used for remote health monitoring. Its regulated use can help mitigate the scarcity of hospital beds and reduce exposure to health-care workers and demand of personal protection equipment.

Unconventional secretory processing diversifies neuronal ion channel properties.

N-glycosylation - the sequential addition of complex sugars to adhesion proteins, neurotransmitter receptors, ion channels and secreted trophic factors as they progress through the endoplasmic reticulum and the Golgi apparatus - is one of the most frequent protein modifications. In mammals, most organ-specific N-glycosylation events occur in the brain. Yet, little is known about the nature, function and regulation of N-glycosylation in neurons. Using imaging, quantitative immunoblotting and mass spectrometry, we show that hundreds of neuronal surface membrane proteins are core-glycosylated, resulting in the neuronal membrane displaying surprisingly high levels of glycosylation profiles that are classically associated with immature intracellular proteins. We report that while N-glycosylation is generally required for dendritic development and glutamate receptor surface expression, core-glycosylated proteins are sufficient to sustain these processes, and are thus functional. This atypical glycosylation of surface neuronal proteins can be attributed to a bypass or a hypo-function of the Golgi apparatus. Core-glycosylation is regulated by synaptic activity, modulates synaptic signaling and accelerates the turnover of GluA2-containing glutamate receptors, revealing a novel mechanism that controls the composition and sensing properties of the neuronal membrane.

Estimation of salivary nitric oxide levels in children with congenital heart diseases.

INTRODUCTION: Oral health problems, especially dental caries are known to be common in children with severe heart disease, especially at early ages. The role of nitric oxide (NO) in protection against oral diseases owing to its strong antimicrobial action has been a topic of interest. OBJECTIVE: The current study was carried out to estimate the NO levels in the salivary samples of children diagnosed with congenital heart diseases (CHDs) and to determine whether salivary NO can serve as a potential biomarker for the evaluation of dental caries-risk. METHODOLOGY: This prospective study was performed on 50 children suffering from CHDs who reported to the department of pediatrics and the levels were compared with those of 50 healthy caries-free children. NO levels in the collected salivary levels were determined spectrophotometrically using the Griess reaction method. RESULTS: Subjects between the age group of 6 and 12 years had maximum decayed teeth (20.95%). The mean NO level in children in study group (60.63) was decreased in comparison to the control group (132.40), which was statistically significant (P value of 0.001). The mean age of the subjects in the study group was 7.24 and the salivary NO levels decreased with age, which was statistically significant. CONCLUSION: Children are suffering from medically compromising conditions such as CHDs are at a higher risk for dental caries and decreased salivary NO levels in these patients may prove as a critical indicator of the same. Saliva can be further utilized as an important biomarker for the determination of the relationship between salivary NO levels and development of dental caries.

Perceptions of Indian dental hygiene students toward their profession and its relationship with their explicit self-esteem scores.

BACKGROUND: The purpose of this study was to investigate the perceptions toward the profession, the level of explicit self-esteem (ESE) of Indian students pursuing the course of dental hygienists, to evaluate the relationship between the two and to develop educational strategies to positively influence students' perceptions. We also wished to evaluate the level of satisfaction of the students to the current status of professional employment in the country. MATERIALS AND METHODS: Students in the second year of the dental hygienist 2-year course were asked to participate in a cross-sectional survey study. An instrument was used to obtain students' perceptions about the profession by estimating the dimensions of "Motivation," "Expectation" and "Environment". Their self-esteem was evaluated using the Rosenberg self-esteem scale. Relationship of self esteem scores with perceptions towards profession was then evaluated. RESULTS: Scores for dimensions including "Motivation," "Expectation" and "Environment" were significantly high, as were the self-esteem scores. The level of ESE was positively correlated with their perceptions of the profession. CONCLUSIONS: The perception of the Indian dental hygienist students was significantly high and positively correlated to the ESE scores. We also conclude that environmental factors may be more influential than innate cultural factors for the development of self-esteem.