Hits and misses with animal models of narcolepsy and the implications for drug discovery.

INTRODUCTION: Narcolepsy is a chronic and rare neurological disorder characterized by disordered sleep. Based on animal models and further research in humans, the dysfunctional orexin system was identified as a contributing factor to the pathophysiology of narcolepsy. Animal models played a larger role in the discovery of some of the pharmacological agents with established benefit/risk profiles. AREAS COVERED: In this review, the authors examine the phenotypes observed in animal models of narcolepsy and the characteristics of clinically used pharmacological agents in these animal models. Additionally, the authors compare the effects of clinically used pharmacological agents on the phenotypes in animal models with those observed in narcolepsy patients. EXPERT OPINION: Research in canine and mouse models have linked narcolepsy to the O×R2mutation and orexin deficiency, leading to new diagnostic criteria and a drug development focus. Advancements in pharmacological therapies have significantly improved narcolepsy management, with insights from both clinical experience and from animal models having led to new treatments such as low sodium oxybate and solriamfetol. However, challenges persist in addressing symptoms beyond excessive daytime sleepiness and cataplexy, highlighting the need for further research, including the development of diurnal animal models to enhance understanding and treatment options for narcolepsy.

A Systematic Review on Deep Learning Model in Computer-aided Diagnosis for Anterior Cruciate Ligament Injury.

INTRODUCTION: In developing Computer-Aided Diagnosis (CAD), a Convolutional Neural Network (CNN) has been commonly used as a Deep Learning (DL) model. Although it is still early, DL has excellent potential in implementing computers in medical diagnosis. METHODS: This study reviews the use of DL for Anterior Cruciate Ligament (ACL) tear diagnosis. A comprehensive search was performed in PubMed, Embase, and Web of Science databases from 2018 to 2024. The included study criteria used MRI images to evaluate ACL tears, and the diagnosis of ACL tears was performed using the DL model. We summarized the paper by reporting their model accuracy, model comparison with arthroscopy, and explainable. RESULTS: AI implementation in tabular format; we conclude that many medical professionals believe that arthroscopic diagnosis is the most reliable method for diagnosing ACL tears. However, due to its intrusive treatment, CAD is projected to be able to produce similar outcomes from MRI scan results. To gain the trust of physicians and meet the demand for reliable knee injury detection systems, an algorithm for CAD should also meet several criteria, such as being transparent, interpretable, explainable, and easy to use. Therefore, future works should consider creating an Explainable DL model for ACL tear diagnosis. It is also essential to evaluate the performance of this Explainable DL model compared to the gold standard of arthroscopy diagnosis. CONCLUSION: There are issues regarding the need for Explainable DL in CAD to increase confidence in its result while also highlighting the importance of the involvement of medical practitioners in system design. There is no funding for this work.

Continuous patient state attention model for addressing irregularity in electronic health records.

BACKGROUND: Irregular time series (ITS) are common in healthcare as patient data is recorded in an electronic health record (EHR) system as per clinical guidelines/requirements but not for research and depends on a patient's health status. Due to irregularity, it is challenging to develop machine learning techniques to uncover vast intelligence hidden in EHR big data, without losing performance on downstream patient outcome prediction tasks. METHODS: In this paper, we propose Perceiver, a cross-attention-based transformer variant that is computationally efficient and can handle long sequences of time series in healthcare. We further develop continuous patient state attention models, using Perceiver and transformer to deal with ITS in EHR. The continuous patient state models utilise neural ordinary differential equations to learn patient health dynamics, i.e., patient health trajectory from observed irregular time steps, which enables them to sample patient state at any time. RESULTS: The proposed models' performance on in-hospital mortality prediction task on PhysioNet-2012 challenge and MIMIC-III datasets is examined. Perceiver model either outperforms or performs at par with baselines, and reduces computations by about nine times when compared to the transformer model, with no significant loss of performance. Experiments to examine irregularity in healthcare reveal that continuous patient state models outperform baselines. Moreover, the predictive uncertainty of the model is used to refer extremely uncertain cases to clinicians, which enhances the model's performance. Code is publicly available and verified at https://codeocean.com/capsule/4587224 . CONCLUSIONS: Perceiver presents a computationally efficient potential alternative for processing long sequences of time series in healthcare, and the continuous patient state attention models outperform the traditional and advanced techniques to handle irregularity in the time series. Moreover, the predictive uncertainty of the model helps in the development of transparent and trustworthy systems, which can be utilised as per the availability of clinicians.

Corrigendum: Sensor-based precision nutrient and irrigation management enhances the physiological performance, water productivity, and yield of soybean under system of crop intensification.

[This corrects the article DOI: 10.3389/fpls.2023.1282217.].

ICEEMDAN-Informer-GWO: a hybrid model for accurate wind speed prediction.

Extensive research has been diligently conducted on wind energy technologies in response to pressing global environmental challenges and the growing demand for energy. Accurate wind speed predictions are crucial for the effective integration of large wind power systems. This study presents a novel and hybrid framework called ICEEMDAN-Informer-GWO, which combines three components to enhance the accuracy of wind speed predictions. The improved complete ensemble empirical mode decomposition with adaptive noise (ICEEMDAN) component improves the decomposition of wind speed data, the Informer model provides computationally efficient wind speed predictions, and the grey wolf optimisation (GWO) algorithm optimises the parameters of the Informer model to achieve superior performance. Three different sets of wind speed prediction (WSP) models and wind farm data from Block Island, Gulf Coast, and Garden City are used to thoroughly assess the proposed hybrid framework. This evaluation focusses on WSP for three specific time horizons: 5 minutes, 30 minutes, and 1 hour ahead. The results obtained from the three conducted experiments conclusively demonstrate that the proposed hybrid framework exhibits superior performance, leading to statistically significant improvements across all three time horizons.

Valorisation of Agri-Food Waste for Bioactive Compounds: Recent Trends and Future Sustainable Challenges.

Worldwide, a massive amount of agriculture and food waste is a major threat to the environment, the economy and public health. However, these wastes are important sources of phytochemicals (bioactive), such as polyphenols, carotenoids, carnitine, coenzymes, essential oils and tocopherols, which have antioxidant, antimicrobial and anticarcinogenic properties. Hence, it represents a promising opportunity for the food, agriculture, cosmetics, textiles, energy and pharmaceutical industries to develop cost effective strategies. The value of agri-food wastes has been extracted from various valuable bioactive compounds such as polyphenols, dietary fibre, proteins, lipids, vitamins, carotenoids, organic acids, essential oils and minerals, some of which are found in greater quantities in the discarded parts than in the parts accepted by the market used for different industrial sectors. The value of agri-food wastes and by-products could assure food security, maintain sustainability, efficiently reduce environmental pollution and provide an opportunity to earn additional income for industries. Furthermore, sustainable extraction methodologies like ultrasound-assisted extraction, pressurized liquid extraction, supercritical fluid extraction, microwave-assisted extraction, pulse electric field-assisted extraction, ultrasound microwave-assisted extraction and high hydrostatic pressure extraction are extensively used for the isolation, purification and recovery of various bioactive compounds from agri-food waste, according to a circular economy and sustainable approach. This review also includes some of the critical and sustainable challenges in the valorisation of agri-food wastes and explores innovative eco-friendly methods for extracting bioactive compounds from agri-food wastes, particularly for food applications. The highlights of this review are providing information on the valorisation techniques used for the extraction and recovery of different bioactive compounds from agricultural food wastes, innovative and promising approaches. Additionally, the potential use of these products presents an affordable alternative towards a circular economy and, consequently, sustainability. In this context, the encapsulation process considers the integral and sustainable use of agricultural food waste for bioactive compounds that enhance the properties and quality of functional food.

A Pillar/Perfusion Plate Enhances Cell Growth, Reproducibility, Throughput, and User Friendliness in Dynamic 3D Cell Culture.

Static three-dimensional (3D) cell culture has been demonstrated in ultralow attachment well plates, hanging droplet plates, and microtiter well plates with hydrogels or magnetic nanoparticles. Although it is simple, reproducible, and relatively inexpensive, thus potentially used for high-throughput screening, statically cultured 3D cells often suffer from a necrotic core due to limited nutrient and oxygen diffusion and waste removal and have a limited in vivo-like tissue structure. Here, we overcome these challenges by developing a pillar/perfusion plate platform and demonstrating high-throughput, dynamic 3D cell culture. Cell spheroids were loaded on the pillar plate with hydrogel by simple sandwiching and encapsulation and cultured dynamically in the perfusion plate on a digital rocker. Unlike traditional microfluidic devices, fast flow velocity was maintained within perfusion wells and the pillar plate was separated from the perfusion plate for cell-based assays. It was compatible with common lab equipment and allowed cell culture, testing, staining, and imaging in situ. The pillar/perfusion plate enhanced cell growth by rapid diffusion, reproducibility, assay throughput, and user friendliness in a dynamic 3D cell culture.

Regenerative human liver organoids (HLOs) in a pillar/perfusion plate for hepatotoxicity assays.

Human liver organoids (HLOs) differentiated from embryonic stem cells (ESCs), induced pluripotent stem cells (iPSCs), and adult stem cells (ASCs) can recapitulate structure and function of human fetal liver tissues, thus, considered as a promising tissue model for liver diseases and predictive compound screening. Nonetheless, there are still several technical challenges to adopt HLOs in the drug discovery process, which include relatively long-term cell differentiation with multiple culture media (3 - 4 weeks) leading to batch-to-batch variation, short-term hepatic function after maturation (3 - 5 days), low assay throughput due to Matrigel dissociation and HLO transfer to a microtiter well plate, and insufficient maturity as compared to primary hepatocytes. To address these issues, expandable HLOs (Exp-HLOs) derived from human iPSCs were generated by optimizing differentiation protocols, which were rapidly printed on a 144-pillar plate with sidewalls and slits (144PillarPlate) and dynamically cultured for up to 20 days into differentiated HLOs (Diff-HLOs) in a 144-perfusion plate with perfusion wells and reservoirs (144PerfusionPlate) for in situ organoid culture and analysis. Dynamically cultured Diff-HLOs were generated robustly and reproducibly in the pillar/perfusion plate with higher maturity as compared to those in statically cultured HLOs by differentiating Exp-HLOs for 10 days. In addition, Diff-HLOs in the pillar/perfusion plate were tested with acetaminophen and troglitazone for 3 days to assess drug-induced liver injury (DILI) and then incubated in an expansion medium for 10 days to evaluate the recovery of the liver from DILI. The assessment of liver regeneration post injury is critical to understand the mechanism of recovery and determine the threshold drug concentration beyond which there will be a sharp decrease in the liver's regenerative capacity. We envision that bioprinted Diff-HLOs in the pillar/perfusion plate could be used for high-throughput screening (HTS) of hepatotoxic compounds due to short-term differentiation of passage-able Exp-HLOs necessary, stable hepatic function after maturation, high reproducibility, and high throughput with capability of in situ organoid culture, testing, staining, imaging, and analysis.

Reproducible generation of human liver organoids (HLOs) on a pillar plate platform via microarray 3D bioprinting.

Human liver organoids (HLOs) hold significant potential for recapitulating the architecture and function of liver tissues in vivo. However, conventional culture methods of HLOs, forming Matrigel domes in 6-/24-well plates, have technical limitations such as high cost and low throughput in organoid-based assays for predictive assessment of compounds in clinical and pharmacological lab settings. To address these issues, we have developed a unique microarray 3D bioprinting protocol of progenitor cells in biomimetic hydrogels on a pillar plate with sidewalls and slits, coupled with a clear bottom, 384-deep well plate for scale-up production of HLOs. Microarray 3D bioprinting, a droplet-based printing technology, was used to generate a large number of small organoids on the pillar plate for predictive hepatotoxicity assays. Foregut cells, differentiated from human iPSCs, were mixed with Matrigel and then printed on the pillar plate rapidly and uniformly, resulting in coefficient of variation (CV) values in the range of 15-18%, without any detrimental effect on cell viability. Despite utilizing 10-50-fold smaller cell culture volume compared to their counterparts in Matrigel domes in 6-/24-well plates, HLOs differentiated on the pillar plate exhibited similar morphology and superior function, potentially due to rapid diffusion of nutrients and oxygen at the small scale. Day 25 HLOs were robust and functional on the pillar plate in terms of their viability, albumin secretion, CYP3A4 activity, and drug toxicity testing, all with low CV values. From three independent trials of in situ assessment, the IC50 values calculated for sorafenib and tamoxifen were 6.2 ± 1.6 µM and 25.4 ± 8.3 µM, respectively. Therefore, our unique 3D bioprinting and miniature organoid culture on the pillar plate could be used for scale-up, reproducible generation of HLOs with minimal manual intervention for high-throughput assessment of compound hepatotoxicity.

Exploring the influence of invasive weed biochar on the sorption and dissipation dynamics of imazethapyr in sandy loam soil.

The management of invasive weeds on both arable and non-arable land is a vast challenge. Converting these invasive weeds into biochar and using them to control the fate of herbicides in soil could be an effective strategy within the concept of turning waste into a wealth product. In this study, the fate of imazethapyr (IMZ), a commonly used herbicide in various crops, was investigated by introducing such weeds as biochar, i.e., Parthenium hysterophorus (PB) and Lantana camara (LB) in sandy loam soil. In terms of kinetics, the pseudo-second order (PSO) model provided the best fit for both biochar-mixed soils. More IMZ was sorbed onto LB-mixed soil compared to PB-mixed soil. When compared to the control (no biochar), both PB and LB biochars (at concentrations of 0.2% and 0.5%) increased IMZ adsorption, although the extent of this effect varied depending on the dosage and type of biochar. The Freundlich adsorption isotherm provided a satisfactory explanation for IMZ adsorption in soil/soil mixed with biochar, with the adsorption process exhibiting high nonlinearity. The values of Gibb's free energy change (ΔG) were negative for both adsorption and desorption in soil/soil mixed with biochar, indicating that sorption was exothermic and spontaneous. Both types of biochar significantly affect IMZ dissipation, with higher degradation observed in LB-amended soil compared to PB-amended soil. Hence, the findings suggest that the preparation of biochar from invasive weeds and its utilization for managing the fate of herbicides can effectively reduce the residual toxicity of IMZ in treated agroecosystems in tropical and subtropical regions.

Recent Advances in Drug Delivery Systems Targeting Insulin Signalling for the Treatment of Alzheimer's Disease.

Alzheimer's disease (AD) is a complex neurodegenerative disorder characterized by the accumulation of neurofibrillary tangles and amyloid-ß plaques. Recent research has unveiled the pivotal role of insulin signaling dysfunction in the pathogenesis of AD. Insulin, once thought to be unrelated to brain function, has emerged as a crucial factor in neuronal survival, synaptic plasticity, and cognitive processes. Insulin and the downstream insulin signaling molecules are found mainly in the hippocampus and cortex. Some molecules responsible for dysfunction in insulin signaling are GSK-3ß, Akt, PI3K, and IRS. Irregularities in insulin signaling or insulin resistance may arise from changes in the phosphorylation levels of key molecules, which can be influenced by both stimulation and inactivity. This, in turn, is believed to be a crucial factor contributing to the development of AD, which is characterized by oxidative stress, neuroinflammation, and other pathological hallmarks. Furthermore, this route is known to be indirectly influenced by Nrf2, NF-κB, and the caspases. This mini-review delves into the intricate relationship between insulin signaling and AD, exploring how disruptions in this pathway contribute to disease progression. Moreover, we examine recent advances in drug delivery systems designed to target insulin signaling for AD treatment. From oral insulin delivery to innovative nanoparticle approaches and intranasal administration, these strategies hold promise in mitigating the impact of insulin resistance on AD. This review consolidates current knowledge to shed light on the potential of these interventions as targeted therapeutic options for AD.

Dynamic culture of cerebral organoids using a pillar/perfusion plate for the assessment of developmental neurotoxicity.

Despite the potential toxicity of commercial chemicals to the development of the nervous system (known as developmental neurotoxicity or DNT), conventional in vitro cell models have primarily been employed for the assessment of acute neuronal toxicity. On the other hand, animal models used for the assessment of DNT are not physiologically relevant due to the heterogenic difference between humans and animals. In addition, animal models are low-throughput, time-consuming, expensive, and ethically questionable. Recently, human brain organoids have emerged as a promising alternative to assess the detrimental effects of chemicals on the developing brain. However, conventional organoid culture systems have several technical limitations including low throughput, lack of reproducibility, insufficient maturity of organoids, and the formation of the necrotic core due to limited diffusion of nutrients and oxygen. To address these issues and establish predictive DNT models, cerebral organoids were differentiated in a dynamic condition in a unique pillar/perfusion plate, which were exposed to test compounds to evaluate DNT potential. The pillar/perfusion plate facilitated uniform, dynamic culture of cerebral organoids with improved proliferation and maturity by rapid, bidirectional flow generated on a digital rocker. Day 9 cerebral organoids in the pillar/perfusion plate were exposed to ascorbic acid (DNT negative) and methylmercury (DNT positive) in a dynamic condition for 1 and 3 weeks, and changes in organoid morphology and neural gene expression were measured to determine DNT potential. As expected, ascorbic acid didn't induce any changes in organoid morphology and neural gene expression. However, exposure of day 9 cerebral organoids to methylmercury resulted in significant changes in organoid morphology and neural gene expression. Interestingly, methylmercury did not induce adverse changes in cerebral organoids in a static condition, thus highlighting the importance of dynamic organoid culture in DNT assessment.

System of wheat intensification (SWI): Effects on lodging resistance, photosynthetic efficiency, soil biomes, and water productivity.

Intense cultivation with narrow row spacing in wheat, a common practice in the Indo-Gangetic plains of South Asia, renders the crop more susceptible to lodging during physiological maturity. This susceptibility, compounded by the use of traditional crop cultivars, has led to a substantial decline in overall crop productivity. In response to these challenges, a two-year field study on the system of wheat intensification (SWI) was conducted. The study involved three different cultivation methods in horizontal plots and four wheat genotypes in vertical plots, organized in a strip plot design. Our results exhibited that adoption of SWI at 20 cm × 20 cm resulted in significantly higher intercellular CO2 concentration (5.9-6.3%), transpiration rate (13.2-15.8%), stomatal conductance (55-59%), net photosynthetic rate (126-160%), and photosynthetically active radiation (PAR) interception (1.6-25.2%) over the existing conventional method (plant geometry 22.5 cm × continuous plant to plant spacing) of wheat cultivation. The lodging resistance capacity of both the lower and upper 3rd nodes was significantly higher in the SWI compared to other cultivation methods. Among different genotypes, HD 2967 demonstrated the highest recorded value for lodging resistance capacity, followed by HD 2851, HD 3086, and HD 2894. In addition, adoption of the SWI at 20 cm × 20 cm enhanced crop grain yield by 36.9-41.6%, and biological yield by 27.5-29.8%. Significantly higher soil dehydrogenase activity (12.06 µg TPF g-1 soil hr-1), arylsulfatase activity (82.8 µg p-nitro phenol g-1 soil hr-1), alkaline phosphatase activity (3.11 n moles ethylene g-1 soil hr-1), total polysaccharides, soil microbial biomass carbon, and soil chlorophyll content were also noted under SWI over conventional method of the production. Further, increased root volumes, surface root density and higher NPK uptake were recorded under SWI at 20×20 cm in comparison to rest of the treatments. Among the tested wheat genotypes, HD-2967 and HD-3086 had demonstrated notable increases in grain and biological yields, as well as improvements in the photosynthetically active radiation (PAR) and chlorophyll content. Therefore, adoption of SWI at 20 cm ×20 cm (square planting) with cultivars HD 2967 might be the best strategy for enhancing crop productivity and resource-use efficiency under the similar wheat growing conditions of India and similar agro-ecotypes of the globe.

Reproducible generation of human liver organoids (HLOs) on a pillar plate platform via microarray 3D bioprinting.

Human liver organoids (HLOs) hold significant potential for recapitulating the architecture and function of liver tissues in vivo. However, conventional culture methods of HLOs, forming Matrigel domes in 6-/24-well plates, have technical limitations such as high cost and low throughput in organoid-based assays for predictive assessment of compounds in clinical and pharmacological lab settings. To address these issues, we have developed a unique microarray 3D bioprinting protocol of progenitor cells in biomimetic hydrogels on a pillar plate with sidewalls and slits, coupled with a clear bottom, 384-deep well plate for scale-up production of HLOs. Microarray 3D bioprinting, a droplet-based printing technology, was used to generate a large number of small organoids on the pillar plate for predictive hepatotoxicity assays. Foregut cells, differentiated from human iPSCs, were mixed with Matrigel and then printed on the pillar plate rapidly and uniformly, resulting in coefficient of variation (CV) values in the range of 15 - 18%, without any detrimental effect on cell viability. Despite utilizing 10 - 50-fold smaller cell culture volume compared to their counterparts in Matrigel domes in 6-/24-well plates, HLOs differentiated on the pillar plate exhibited similar morphology and superior function, potentially due to rapid diffusion of nutrients and oxygen at the small scale. Day 25 HLOs were robust and functional on the pillar plate in terms of their viability, albumin secretion, CYP3A4 activity, and drug toxicity testing, all with low CV values. From three independent trials of in situ assessment, the IC50 values calculated for sorafenib and tamoxifen were 6.2 ± 1.6 µM and 25.4 ± 8.3 µM, respectively. Therefore, our unique 3D bioprinting and miniature organoid culture on the pillar plate could be used for scale-up, reproducible generation of HLOs with minimal manual intervention for high-throughput assessment of compound hepatotoxicity.

A Multicentric, Prospective, Observational, Single-Arm Registry Study to Assess the Clinical Safety and Effectiveness of Thrombophob Ointment (Heparin Sodium + Benzyl Nicotinate) in Indian Patients With Thrombophlebitis.

Purpose Thrombophlebitis is a frequent intravenous (IV) therapy consequence. Topical heparin for seven days is used as a treatment for thrombophlebitis. This study was performed to evaluate the clinical safety and effectiveness of the combination of heparin sodium & benzyl nicotinate (Thrombophob Ointment, manufactured by Zydus Healthcare Ltd., Ahmedabad, India) in thrombophlebitis patients in India. Methods A study carried out by 118 Indian doctors examined 2002 thrombophlebitis patients from 2016-2023, prescribing ointment containing heparin sodium and benzyl nicotinate. Patients were followed up on day three and day seven after starting the treatment, and safety and effectiveness were recorded, including adverse events. Result A total of 2002 patients were included in the study and males were predominant (58.15%). IV fluids (60.58%) were the leading cause of thrombophlebitis. The study found notable improvements in key markers of venous health over time. Compared to baseline, patients experienced significantly reduced severity of phlebitis, shorter venous lesion lengths, and lower pain and tenderness scores by both day 3 and day 7 (p<0.001 for all comparisons). Furthermore, these improvements continued between day 3 and day 7, indicating sustained positive effects (p<0.001 for all comparisons). After the application of the ointment, very few patients experienced adverse effects (0.25% on day three and 0.05% on day seven). Treatment effectiveness was excellent in 72% of patients, and treatment safety was excellent in 93% of patients. Conclusion The ointment containing heparin sodium and benzyl nicotinate was well tolerated and efficacious in the treatment of thrombophlebitis in Indian patients.

Thermostable bacterial L-asparaginase for polyacrylamide inhibition and in silico mutational analysis.

The L-asparaginase (ASPN) enzyme has received recognition in various applications including acrylamide degradation in the food industry. The synthesis and application of thermostable ASPN enzymes is required for its use in the food sector, where thermostable enzymes can withstand high temperatures. To achieve this goal, the bacterium Bacillus subtilis was isolated from the hot springs of Tapovan for screening the production of thermostable ASPN enzyme. Thus, ASPN with a maximal specific enzymatic activity of 0.896 U/mg and a molecular weight of 66 kDa was produced from the isolated bacteria. The kinetic study of the enzyme yielded a Km value of 1.579 mM and a Vmax of 5.009 µM/min with thermostability up to 100 min at 75 °C. This may have had a positive indication for employing the enzyme to stop polyacrylamide from being produced. The current study has also been extended to investigate the interaction of native and mutated ASPN enzymes with acrylamide. This concluded that the M10 (with 10 mutations) has the highest protein and thermal stability compared to the wild-type ASPN protein sequence. Therefore, in comparison to a normal ASPN and all other mutant ASPNs, M10 is the most favorable mutation. This research has also demonstrated the usage of ASPN in food industrial applications.

Differential host responses to COVID-19: Unraveling the complexity.

These diverse outcomes of Covid-19 are influenced by various factors including age, gender, underlying health conditions, immune responses, viral variants, external factors, and overall quality of life. Demographic analysis of patients aged 0-18 years experienced mild to moderate cases, above 55 years with co-morbidities, were more severely affected.COVID-19 incidence was higher in males (58 %) & (42 %) in females. The reduced expression of Toll-like receptors (TLR) in severe and critical patients is a crucial determinant. This reduced TLR expression is primarily attributed to the dominance of the PLpro viral protein of COVID-19. Disease enrichment analysis highlights the long-term impact of COVID-19, which can lead to post-recovery complications such as hypertension, diabetes, cardiac diseases, and brain ischemia in Covid-19 patients. In conclusion, a comprehensive strategy targeting key factors like PLpro, TLR, and inflammatory cytokines such as IL-1 and IL-6 could offer an effective approach to mitigate the devastating effects of COVID-19.

The long intergenic non-coding RNA LINC01140 modulates gastric cancer phenotypes and cancer cell lines aggressiveness.

BACKGROUND: Long-intergenic non-protein coding gene 01140 (LINC01140) a long non-coding RNA is highly expressed in various cancers. However, its biological functions in gastric cancer progression is still unknown. METHOD: To elucidate LINC01140 function, 70 GC tumor samples and 30 normal gastric tissues were collected. LINC01140 expression level were determined by qRT-PCR analysis and correlated with different clinico-pathological parameters. Then we tried to see the impact of LINC01140 on gastric cell line aggressiveness by knocking down the target gene and performing cell viability assay, migration assay and invasive capacity of the cell lines along with immunoblotting to check several protein levels. RESULT: LINC01140 RNA is found to be positively correlated with FGF9 and significantly up regulated in GC tissues. LINC01140 knockdown inhibited the viability, migratory capacity and invasive capacity of AGS cells. LINC01140 targets miR-140-5p, while miR-140-5p targeted FGF9 to form lncRNA-miRNA-mRNA axis. The affect of miR-140-5p inhibition on gastric cancer cell aggressiveness were opposite to those of LINC01140 or FGF9 knockdown. Additionally, inhibition partially reversed the effects of LINC01140 knockdown on FGF9 protein levels, gastric cancer cell phenotypes. CONCLUSION: LINC01140, miR-140-5p and FGF9 form a lncRNA-miRNA-mRNA axis that modulates the gastric cancer phenotypes and in turn affects gastric cancer cell aggressiveness.

Cryo-electron microscopy-based drug design.

Structure-based drug design (SBDD) has gained popularity owing to its ability to develop more potent drugs compared to conventional drug-discovery methods. The success of SBDD relies heavily on obtaining the three-dimensional structures of drug targets. X-ray crystallography is the primary method used for solving structures and aiding the SBDD workflow; however, it is not suitable for all targets. With the resolution revolution, enabling routine high-resolution reconstruction of structures, cryogenic electron microscopy (cryo-EM) has emerged as a promising alternative and has attracted increasing attention in SBDD. Cryo-EM offers various advantages over X-ray crystallography and can potentially replace X-ray crystallography in SBDD. To fully utilize cryo-EM in drug discovery, understanding the strengths and weaknesses of this technique and noting the key advancements in the field are crucial. This review provides an overview of the general workflow of cryo-EM in SBDD and highlights technical innovations that enable its application in drug design. Furthermore, the most recent achievements in the cryo-EM methodology for drug discovery are discussed, demonstrating the potential of this technique for advancing drug development. By understanding the capabilities and advancements of cryo-EM, researchers can leverage the benefits of designing more effective drugs. This review concludes with a discussion of the future perspectives of cryo-EM-based SBDD, emphasizing the role of this technique in driving innovations in drug discovery and development. The integration of cryo-EM into the drug design process holds great promise for accelerating the discovery of new and improved therapeutic agents to combat various diseases.