Von Hippel Lindau tumor suppressor controls m6A-dependent gene expression in renal tumorigenesis.

N6-Methyladenosine (m6A) is the most abundant posttranscriptional modification, and its contribution to cancer evolution has recently been appreciated. Renal cancer is the most common adult genitourinary cancer, approximately 85% of which is accounted for by the clear cell renal cell carcinoma (ccRCC) subtype characterized by VHL loss. However, it is unclear whether VHL loss in ccRCC affects m6A patterns. In this study, we demonstrate that VHL binds and promotes METTL3/METTL14 complex formation while VHL depletion suppresses m6A modification, which is distinctive from its canonical E3 ligase role. m6A RNA immunoprecipitation sequencing (RIP-Seq) coupled with RNA-Seq allows us to identify a selection of genes whose expression may be regulated by VHL-m6A signaling. Specifically, PIK3R3 is identified to be a critical gene whose mRNA stability is regulated by VHL in a m6A-dependent but HIF-independent manner. Functionally, PIK3R3 depletion promotes renal cancer cell growth and orthotopic tumor growth while its overexpression leads to decreased tumorigenesis. Mechanistically, the VHL-m6A-regulated PIK3R3 suppresses tumor growth by restraining PI3K/AKT activity. Taken together, we propose a mechanism by which VHL regulates m6A through modulation of METTL3/METTL14 complex formation, thereby promoting PIK3R3 mRNA stability and protein levels that are critical for regulating ccRCC tumorigenesis.

Microwave-assisted ZnO-decorated carbon urchin resembling 'shish-kebab' morphology with self-healing and enhanced electromagnetic shielding properties.

Herein, inspired by Acacia auriculiformis fruit, the shish-kebab-like growth of ZnO on carbon urchin (ZnO@CU) was designed using microwave radiation, thus leading to a hierarchal 3D structure that can promote multiple internal reflections through polarization centers. This hierarchal structure was then dispersed in a designer polyetherimide (PEI) matrix containing dynamic covalent bonds that can undergo metathesis, triggered by temperature, to harness self-healing properties in the composite. Such key attributes are required for their potential use in EMI shielding applications where frequent repairs are indispensable. Morphological investigation revealed that the ZnO flower was periodically nucleated like 'shish-kebab' structures on CU surfaces. CU was designed from short carbon fibers using a facile modified method. The EMI shielding performance of the resulting composites was investigated in the X-band, illustrating a shielding effectiveness of -40.6 dB for 2 wt% of ZnO@CU loading, and the composite can be preserved after the self-healing procedure. The ZnO 'kebabs' on 'CU shish' facilitated multiple scattering and numerous polarization centers to improve the EMI shielding performances at extremely low filler contents. In addition, the mechanical and thermal properties of the composite showed improved structural integrity and superior resistance to extreme temperatures, respectively. Overall, the proposed ZnO@CU/PEI composite has great potential to fulfill the increasing demands for lightweight EMI shielding materials in many fields.

Advances in RNA Interference for Plant Functional Genomics: Unveiling Traits, Mechanisms, and Future Directions.

RNA interference (RNAi) is a conserved molecular mechanism that plays a critical role in post-transcriptional gene silencing across diverse organisms. This review delves into the role of RNAi in plant functional genomics and its applications in crop improvement, highlighting its mechanistic insights and practical implications. The review begins with the foundational discovery of RNAi's mechanism, tracing its origins from petunias to its widespread presence in various organisms. Various classes of regulatory non-coding small RNAs, including siRNAs, miRNAs, and phasiRNAs, have been uncovered, expanding the scope of RNAi-mediated gene regulation beyond conventional understanding. These RNA classes participate in intricate post-transcriptional and epigenetic processes that influence gene expression. In the context of crop enhancement, RNAi has emerged as a powerful tool for understanding gene functions. It has proven effective in deciphering gene roles related to stress resistance, metabolic pathways, and more. Additionally, RNAi-based approaches hold promise for integrated pest management and sustainable agriculture, contributing to global efforts in food security. This review discusses RNAi's diverse applications, such as modifying plant architecture, extending shelf life, and enhancing nutritional content in crops. The challenges and future prospects of RNAi technology, including delivery methods and biosafety concerns, are also explored. The global landscape of RNAi research is highlighted, with significant contributions from regions such as China, Europe, and North America. In conclusion, RNAi remains a versatile and pivotal tool in modern plant research, offering novel avenues for understanding gene functions and improving crop traits. Its integration with other biotechnological approaches such as gene editing holds the potential to shape the future of agriculture and sustainable food production.

Dataset for boiling acoustic emissions: A tool for data driven boiling regime prediction.

Boiling is used for the thermal management of high-energy-density devices and systems. However, sudden thermal runaway at boiling crisis often results in catastrophic failures. Machine learning is a promising tool for in-situ monitoring of boiling-based systems for preemptive control of boiling crisis. A carefully acquired and well-labeled dataset is a primary requirement for utilizing any data-driven learning framework to extract valuable descriptors. Here, we present a comprehensive dataset of boiling acoustics presented in our recent work [1]. We collect the audio files through meticulously controlled near-saturated pool boiling experiments under steady-state conditions. To this end, we connect a high-sensitivity hydrophone to a pre-amplifier and a data acquisition unit for accurate and reliable acquisition of acoustic signals. We organize the audio files into four categories as per the respective boiling regimes: background or natural convection (BKG, 2-5W/cm2), nucleate boiling (NB, 8-140W/cm2), excluding those at higher heat flux values preceding the onset of boiling crisis or the critical heat flux (Pre-CHF, ≈145W/cm2), and transition boiling (TB, uncontrolled). Each audio file label provides explicit information about the heat flux value and the experimental conditions. This dataset, consisting of 2056 files for BKG, 13367 files for NB, 399 files for Pre-CHF, and 460 files for TB, serves as the foundation for training and evaluating a deep learning strategy to predict boiling regimes. The dataset also includes acoustic emission data from transient pool boiling experiments conducted with varying heating strategies, heater surface, and boiling fluid modifications, creating a valuable dataset for developing robust data-driven models to predict boiling regimes. We also provide the associated MATLAB® codes used to process and classify these audio files.

Investigating the Vapor-Phase Adsorption of Aroma Molecules on the Water-Vapor Interface using Molecular Dynamics Simulations.

Surfactants are amphiphilic additives primarily used to reduce the surface tension of water and manipulate its wettability on various surfaces. Recent reports suggest that volatile surfactants, such as aroma molecules, diffuse more quickly to the interface from the vapor-phase than conventional surfactants typically used in the aqueous phase. The ability to adsorb from the vapor phase, in addition to their use as cosurfactants, expands the potential applications of volatile surfactants, particularly in situations where adding surfactants from the liquid phase is difficult. Here, we present a molecular level understanding of the adsorption kinetics of linalool, a common aroma molecule, on the water interface using molecular dynamics simulations. We note that the value of surface tension while adsorption from vapor and liquid phases is dependent only on the surface coverage. A minimum surface tension of 32 ± 1.8 mN/m is obtained in both cases at a maximum surface coverage of 4.88 µmol/m2 at 300 K. We observe the extent of decrease of the H-bonds between linalool-water and linalool-linalool molecules at various surface coverages to explain the mechanism of surface tension reduction. We solve Gibb's adsorption equation to establish a correlation between the surface coverage of linalool and the corresponding bulk concentration in experiments. We investigate the free energy profile of linalool's adsorption behavior at different surface coverages and temperatures. Our report suggests that linalool adsorption onto the water interface is an enthalpy-driven process primarily dependent on the strength of the interaction between the hydroxyl group of linalool and water molecules. These insights are crucial for selecting a suitable aroma molecule for various applications that target the vapor-phase adsorption mechanism.

Gut microbiome supplementation as therapy for metabolic syndrome.

The gut microbiome is defined as an ecological community of commensal symbiotic and pathogenic microorganisms that exist in our body. Gut microbiome dysbiosis is a condition of dysregulated and disrupted intestinal bacterial homeostasis, and recent evidence has shown that dysbiosis is related to chronic inflammation, insulin resistance, cardiovascular diseases (CVD), type 2 diabetes mellitus (T2DM), and obesity. It is well known that obesity, T2DM and CVD are caused or worsened by multiple factors like genetic predisposition, environmental factors, unhealthy high calorie diets, and sedentary lifestyle. However, recent evidence from human and mouse models suggest that the gut microbiome is also an active player in the modulation of metabolic syndrome, a set of risk factors including obesity, hyperglycemia, and dyslipidemia that increase the risk for CVD, T2DM, and other diseases. Current research aims to identify treatments to increase the number of beneficial microbiota in the gut microbiome in order to modulate metabolic syndrome by reducing chronic inflammation and insulin resistance. There is increasing interest in supplements, classified as prebiotics, probiotics, synbiotics, or postbiotics, and their effect on the gut microbiome and metabolic syndrome. In this review article, we have summarized current research on these supplements that are available to improve the abundance of beneficial gut microbiota and to reduce the harmful ones in patients with metabolic syndrome.

AutoML accurately predicts endovascular mechanical thrombectomy in acute large vessel ischemic stroke.

Background and objective: Automated machine learning or autoML has been widely deployed in various industries. However, their adoption in healthcare, especially in clinical settings is constrained due to a lack of clear understanding and explainability. The aim of this study is to utilize autoML for the prediction of functional outcomes in patients who underwent mechanical thrombectomy and compare it with traditional ML models with a focus on the explainability of the trained models. Methods: A total of 156 patients of acute ischemic stroke with Large Vessel Occlusion (LVO) who underwent mechanical thrombectomy within 24 h of stroke onset were included in the study. A total of 34 treatment variables including clinical, demographic, imaging, and procedure-related data were extracted. Various conventional machine learning models such as decision tree classifier, logistic regression, random forest, kNN, and SVM as well as various autoML models such as AutoGluon, MLJAR, Auto-Sklearn, TPOT, and H2O were used to predict the modified Rankin score (mRS) at the time of patient discharge and 3 months follow-up. The sensitivity, specificity, accuracy, and AUC for traditional ML and autoML models were compared. Results: The autoML models outperformed the traditional ML models. For the prediction of mRS at discharge, the highest testing accuracy obtained by traditional ML models for the decision tree classifier was 74.11%, whereas for autoML which was obtained through AutoGluon, it showed an accuracy of 88.23%. Similarly, for mRS at 3 months, the highest testing accuracy of traditional ML was that of the SVM classifier at 76.5%, whereas that of autoML was 85.18% obtained through MLJAR. The 24-h ASPECTS score was the most important predictor for mRS at discharge whereas for prediction of mRS at 3 months, the most important factor was mRS at discharge. Conclusion: Automated machine learning models based on multiple treatment variables can predict the functional outcome in patients more accurately than traditional ML models. The ease of clinical coding and deployment can assist clinicians in the critical decision-making process. We have developed a demo application which can be accessed at https://mrs-score-calculator.onrender.com/.

Lung function trajectories in patients with idiopathic pulmonary fibrosis.

BACKGROUND: Idiopathic pulmonary fibrosis (IPF) is a progressive fibrosing interstitial lung disease characterised by decline in lung function. We evaluated trajectories of forced vital capacity (FVC) and diffusing capacity (DLco) in a cohort of patients with IPF. METHODS: Patients with IPF that was diagnosed or confirmed at the enrolling centre in the previous 6 months were enrolled into the IPF-PRO Registry between June 2014 and October 2018. Patients were followed prospectively, with lung function data collected as part of routine clinical care. Mean trajectories of FVC and DLco % predicted in all patients and in subgroups by characteristics assessed at enrolment were estimated using a joint model that accounted for factors such as disease severity and visit patterns. RESULTS: Of 1002 patients in the registry, 941 had ≥ 1 FVC and/or DLco measurement after enrolment. The median (Q1, Q3) follow-up period was 35.1 (18.9, 47.2) months. Overall, mean estimated declines in FVC and DLco % predicted were 2.8% and 2.9% per year, respectively. There was no evidence that the mean trajectories of FVC or DLco had a non-linear relationship with time at the population level. Patients who were male, white, had a family history of ILD, were using oxygen, or had prior/current use of antifibrotic therapy at enrolment had greater rates of decline in FVC % predicted. Patients who were male or white had greater rates of decline in DLco % predicted. CONCLUSIONS: Data from the IPF-PRO Registry suggest a constant rate of decline in lung function over a prolonged period, supporting the inexorably progressive nature of IPF. A graphical abstract summarising the data in this manuscript is available at: https://www.usscicomms.com/respiratory/IPF-PRORegistry_LungFunctionTrajectories . TRIAL REGISTRATION: NCT01915511.

A double-blind, placebo-controlled, randomized withdrawal trial of sarilumab for the treatment of glucocorticoid-dependent sarcoidosis.

OBJECTIVES: Effective steroid-sparing therapies for the treatment of sarcoidosis are lacking; interleukin-6 (IL-6) antagonists may reduce sarcoidosis disease activity. This study assessed the safety and efficacy of the IL-6 receptor antagonist, sarilumab, in subjects with glucocorticoid-dependent sarcoidosis. METHODS: This phase II, double-blind, placebo-controlled, randomized withdrawal trial enrolled 15 subjects with biopsy-proven sarcoidosis at Stanford University from November 2019 to September 2022. In Period 1, subjects were treated with open-label sarilumab 200mg subcutaneously every two weeks for 16 weeks, with predefined tapering of prednisone. Subjects who completed Period 1 without a sarcoidosis flare entered Period 2 and were randomized to continue sarilumab or to receive matching placebo for 12 weeks. Endpoints included flare-free survival, as well as changes in pulmonary function tests, chest imaging, patient reported outcomes, and laboratory values. RESULTS: Fifteen subjects were enrolled in the study (median age 57 years, 80% male, 73.3% White), and 10 subjects successfully completed Period 1. During Period 1, 4 of 15 subjects (26.7%) discontinued due to worsening of their sarcoidosis, and CT chest imaging worsened in 5 of 15 subjects (35.7%). During Period 2, 0 of 2 subjects in the sarilumab group and 1 of 8 subjects (12.5%) in the placebo group had a flare. Treatment with sarilumab 200 mg was generally well tolerated in subjects with sarcoidosis. CONCLUSION: In this double-blind, placebo-controlled, randomized withdrawal trial, a meaningful signal for improvement in subjects with sarcoidosis treated with sarilumab was not observed. Given the small numbers in this study, no definitive conclusions can be drawn. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT04008069.

Electrophoretically Deposited Multiscale Graphene Oxide/Carbon Nanotube Construct Mediated Interfacial Engineering in Carbon Fiber Epoxy Composites.

Fiber-reinforced polymer composites as a structural material have garnered tremendous interest over the past few decades. In particular, carbon fiber-reinforced epoxy (CFRE) laminates have seen extensive use in the aircraft and aerospace industry. The role of the interface between the matrix and fiber is critical and dictates the overall structural properties of the CFRE laminate. Herein, we attempt to use a commercially viable, green, and facile approach, electrophoretic deposition (EPD), to deposit covalently coupled multiscale graphene oxide (GO)/carbon nanotube (CNT) nanoconstructs onto carbon fiber (CF) fabric. The rationale behind using these hybrid conjugates is to exploit the positive synergistic effect of combining two-dimensional (2D) GO and one-dimensional (1D) CNT nanoparticles, which provide strengthening through different mechanisms resulting in a stronger matrix/fiber interface. The modified laminate with just 0.1 wt % GO/CNT content exhibited an improvement in flexural strength (FS) by 24% and interlaminar shear strength (ILSS) by 30% compared to the neat CFRE. Scanning electron microscope (SEM) micrographs confirmed uniform and homogeneous GO and GO/CNT deposition on CF. Raman, Fourier-transform infrared spectroscopy (FTIR), and X-ray photoelectron spectroscopy (XPS) analyses validate the successful functionalization of CNT and covalent coupling of GO and CNT. Atomic force microscope (AFM) and contact angle analyses indicate improved interaction between the CF and matrix. The deposition of the GO/CNT nanoconstruct on the CF improved the performance of CFREs owing to enhanced wettability, surface free energy, and surface roughness, leading to increased mechanical interlocking between the epoxy and CF at the interface. Dynamic mechanical analysis showed decreased segmental motion of epoxy chains due to improved interfacial adhesion following modification. Interesting observations were made in SEM fractography, which showed considerably different failure mechanisms in the modified CFREs. Electromagnetic interference (EMI) shielding effectiveness of -45 dB was achieved in the case of the GO/CNT-CFRE system. Electrothermal heating and de-icing performance of the modified system were also explored in this study. This versatile approach can open up new avenues for CFRE modification leading to considerably improved performance.

AIMS65, Glasgow-Blatchford bleeding score and modified Glasgow-Blatchford bleeding score in predicting outcomes of upper gastrointestinal bleeding: An accuracy and calibration study.

BACKGROUND: Albumin, international normalized ratio (INR), mental status, systolic blood pressure, age >65 years (AIMS65), Glasgow-Blatchford bleeding score (GBS) and modified GBS (mGBS) are three pre-endoscopy scoring systems used in the risk stratification of upper gastrointestinal bleeding (UGIB). The utility of such scoring systems in a population is estimated by their accuracy and calibration in the population. We aimed at validating and comparing the accuracy of the three scoring systems in predicting clinical outcomes including in-hospital mortality, need for blood transfusion, endoscopic treatment and rebleeding risk. METHOD: We conducted a single-center, retrospective cohort study on patients with UGIB at a tertiary care center in India over 12 months. Clinical and laboratory data was collected from all patients admitted with UGIB. All patients were risk stratified using AIMS65, GBS and mGBS. The clinical outcome examined were: in-hospital mortality, requirement of blood transfusion, need for endoscopic treatment and rebleeding during hospital stay. The area under receiver-operating curve (AUROC) was calculated to assess the performance and calibration curves (Hosmer-Lemeshow goodness of fit curve) were plotted to examine how accurately the model describes the data of all three scoring systems. RESULTS: Total 260 patients were included in the study, of which 236 (90.8%) were males. As many as 144 (55.4%) patients required blood transfusion and 64 (30.8%) required endoscopic treatment. While the incidence of rebleeding was 7.7%, in hospital mortality was 15.4%. Of 208 who underwent endoscopy, the most common causes identified were varices (49%) and gastritis (18.2%), followed by ulcer (11%), Mallory-Weiss tear (8.1%), portal hypertensive gastropathy (6.7%), malignancy (4.8%) and esophageal candidiasis (1.9%). The median AIMS65 score was 1, GBS 7 and mGBS 6. The area under curve (AUROC) for AIMS65, GBS and mGBS was (0.77, 0.73,0.70), (0.75, 0.82,0.83), (0.56, 0.58,0.83), (0.81, 0.94,0.53) for in-hospital mortality, blood transfusion requirement, endoscopic treatment and rebleeding prediction, respectively. CONCLUSION: GBS and mGBS are superior to AIMS65 in predicting the requirement of blood transfusion and rebleeding risk, whereas in-hospital mortality was better predicted by AIMS 65. Both scores performed poorly in predicting the need of endoscopic treatment. An AIMS65 of 0,1 and a GBS of ≤ 1 are not associated with significant adverse events. A poor calibration of the scores in our population points to the lack of generalizability of these scoring systems.

Integrative multi-omics analysis reveals novel idiopathic pulmonary fibrosis endotypes associated with disease progression.

BACKGROUND: Idiopathic pulmonary fibrosis (IPF) is characterized by the accumulation of extracellular matrix in the pulmonary interstitium and progressive functional decline. We hypothesized that integration of multi-omics data would identify clinically meaningful molecular endotypes of IPF. METHODS: The IPF-PRO Registry is a prospective registry of patients with IPF. Proteomic and transcriptomic (including total RNA [toRNA] and microRNA [miRNA]) analyses were performed using blood collected at enrollment. Molecular data were integrated using Similarity Network Fusion, followed by unsupervised spectral clustering to identify molecular subtypes. Cox proportional hazards models tested the relationship between these subtypes and progression-free and transplant-free survival. The molecular subtypes were compared to risk groups based on a previously described 52-gene (toRNA expression) signature. Biological characteristics of the molecular subtypes were evaluated via linear regression differential expression and canonical pathways (Ingenuity Pathway Analysis [IPA]) over-representation analyses. RESULTS: Among 232 subjects, two molecular subtypes were identified. Subtype 1 (n = 105, 45.3%) and Subtype 2 (n = 127, 54.7%) had similar distributions of age (70.1 +/- 8.1 vs. 69.3 +/- 7.6 years; p = 0.31) and sex (79.1% vs. 70.1% males, p = 0.16). Subtype 1 had more severe disease based on composite physiologic index (CPI) (55.8 vs. 51.2; p = 0.002). After adjusting for CPI and antifibrotic treatment at enrollment, subtype 1 experienced shorter progression-free survival (HR 1.79, 95% CI 1.28,2.56; p = 0.0008) and similar transplant-free survival (HR 1.30, 95% CI 0.87,1.96; p = 0.20) as subtype 2. There was little agreement in the distribution of subjects to the molecular subtypes and the risk groups based on 52-gene signature (kappa = 0.04, 95% CI= -0.08, 0.17), and the 52-gene signature risk groups were associated with differences in transplant-free but not progression-free survival. Based on heatmaps and differential expression analyses, proteins and miRNAs (but not toRNA) contributed to classification of subjects to the molecular subtypes. The IPA showed enrichment in pulmonary fibrosis-relevant pathways, including mTOR, VEGF, PDGF, and B-cell receptor signaling. CONCLUSIONS: Integration of transcriptomic and proteomic data from blood enabled identification of clinically meaningful molecular endotypes of IPF. If validated, these endotypes could facilitate identification of individuals likely to experience disease progression and enrichment of clinical trials. TRIAL REGISTRATION: NCT01915511.

Nitrogen use efficiency-a key to enhance crop productivity under a changing climate.

Nitrogen (N) is an essential element required for the growth and development of all plants. On a global scale, N is agriculture's most widely used fertilizer nutrient. Studies have shown that crops use only 50% of the applied N effectively, while the rest is lost through various pathways to the surrounding environment. Furthermore, lost N negatively impacts the farmer's return on investment and pollutes the water, soil, and air. Therefore, enhancing nitrogen use efficiency (NUE) is critical in crop improvement programs and agronomic management systems. The major processes responsible for low N use are the volatilization, surface runoff, leaching, and denitrification of N. Improving NUE through agronomic management practices and high-throughput technologies would reduce the need for intensive N application and minimize the negative impact of N on the environment. The harmonization of agronomic, genetic, and biotechnological tools will improve the efficiency of N assimilation in crops and align agricultural systems with global needs to protect environmental functions and resources. Therefore, this review summarizes the literature on nitrogen loss, factors affecting NUE, and agronomic and genetic approaches for improving NUE in various crops and proposes a pathway to bring together agronomic and environmental needs.

Adverse Events Related to Tirzepatide.

Context: Tirzepatide is a dual glucose-dependent insulinotropic peptide (GIP) and glucagon-like peptide-1 receptor agonist (GLP-1 RA) approved by the US Food and Drug Administration in May 2022 for patients with type 2 diabetes mellitus (T2DM). Objective: We aimed to determine the rates of individual adverse events (AEs) related to 3 studied doses of tirzepatide. Methods: We performed a systematic review with meta-analysis including 5 databases (PubMed, Embase, CINAHL, Scopus, and Web of Science) for all clinical trials reporting AEs related to tirzepatide. The safety data from individual studies were extracted and analyzed through meta-regression to assess rates of individual AEs. Study quality assessment was performed using the National Heart, Lung, and Blood Institute Quality Assessment Tool for Observational Cohort and Cross-Sectional Studies. Results: Ten trials (6836 participants) were included. Gastrointestinal (GI) AEs were the most commonly reported AEs and were dose dependent 39% (95% CI, 35%-43%), 46% (95% CI, 42%-49%), and 49% (95% CI, 38%-60%) for the 5, 10, and 15 mg dose, respectively. Among all GI AEs, nausea and diarrhea were most frequent at any dose of tirzepatide. Drug discontinuation due to AEs was highest with the 15 mg dose of tirzepatide (10%). Incidence of mild hypoglycemia (blood glucose < 70 mg/dL) was highest with tirzepatide 10 mg dose 22.6% (9.2%-39.8%). Rates of fatal AEs, severe hypoglycemia, acute pancreatitis, cholelithiasis, and cholecystitis were extremely low (≤ 1%) across all doses of tirzepatide. Conclusion: Tirzepatide is associated with a dose-dependent increase in incidence of GI AEs and AEs leading to drug discontinuation. Severe hypoglycemia, fatal AEs, acute pancreatitis, cholelithiasis, and cholecystitis are rare with this medication.

Structures and mechanisms of tRNA methylation by METTL1-WDR4.

Specific, regulated modification of RNAs is important for proper gene expression1,2. tRNAs are rich with various chemical modifications that affect their stability and function3,4. 7-Methylguanosine (m7G) at tRNA position 46 is a conserved modification that modulates steady-state tRNA levels to affect cell growth5,6. The METTL1-WDR4 complex generates m7G46 in humans, and dysregulation of METTL1-WDR4 has been linked to brain malformation and multiple cancers7-22. Here we show how METTL1 and WDR4 cooperate to recognize RNA substrates and catalyse methylation. A crystal structure of METTL1-WDR4 and cryo-electron microscopy structures of METTL1-WDR4-tRNA show that the composite protein surface recognizes the tRNA elbow through shape complementarity. The cryo-electron microscopy structures of METTL1-WDR4-tRNA with S-adenosylmethionine or S-adenosylhomocysteine along with METTL1 crystal structures provide additional insights into the catalytic mechanism by revealing the active site in multiple states. The METTL1 N terminus couples cofactor binding with conformational changes in the tRNA, the catalytic loop and the WDR4 C terminus, acting as the switch to activate m7G methylation. Thus, our structural models explain how post-translational modifications of the METTL1 N terminus can regulate methylation. Together, our work elucidates the core and regulatory mechanisms underlying m7G modification by METTL1, providing the framework to understand its contribution to biology and disease.

The comparison of Canopeo and samplepoint for measurement of green canopy cover for forage crops in India.

Canopy covers can be measured using destructive (visual) and non-destructive methods (spectral indices, photogrammetry, visual assessment, and quantum sensor). The precision of crop cover estimation, however, is dependent on the selection of appropriate methods. Studies were conducted at the Indian Grassland and Fodder Research Institute, Jhansi to compare the forage crops canopy cover estimated using photogrammetry software (Canopeo and SamplePoint) and visual assessments. Assessments were performed in three summer crops (corn, cowpea, and sorghum), two winter crops (Egyptian clover, and oats), and bare ground condition. For each plot, three nadir images (directly above the canopy) were captured using digital cameras from a height of 1.5 m above the soil surface between 10 AM to 2 PM on bright sunny days. The results indicated that the relationships between visual assessment and Canopeo (regression coefficient, (R2 = 0.96), visual assessment and SamplePoint (0.96), and Canopeo and SamplePoint (0.98) were linear when data were pooled across all the crops. SamplePoint and Canopeo is further, appropriate for cowpea (Pearson coefficient (R = 0.99 and 0.94), oats (0.92 and 0.97), and sorghum (0.46 and 0.51), respectively. SamplePoint and Canopeo are not suitable for berseem (-0.15) and corn (-0.61), respectively, due to dead residues after the first harvest in berseem and taller corn might have influenced the image quality. Therefore, the stage of the crop, the height of the crop, and dead residues around the plants can greatly influence the estimation of crop cover. In conclusion, the results indicated that this photogrammetry software can be used for non-destructive crop canopy measurement with the above-mentioned precautions in the forage crops tested. •Forage canopy cover is estimated generally by visual scoring, and the outcome varies widely from person to person.•Photogrammetry methods (Canopeo and SamplePoint) were positiviely correlated with visual scoring for cowpea, oats, and sorghum.•However, Canopeo and SamplePoint may not suitable for taller crops like corn and ratoon crops like berseem.

Water, sanitation, and hygiene (WaSH) practices among people living with HIV/AIDS (PLHA) - need to relook, in the era of COVID-19 pandemic.

Background: Water, sanitation, and hygiene (WaSH) practices always have been neglected among HIV/AIDS (Human immunodeficiency virus/ Acquired immunodeficiency syndrome) programs, even when HIV and WaSH services have robust bearing on each other. With COVID-19 pandemic on the go, it is utmost necessary for the people living with HIV/AIDS (PLHA) to ensure adequate WaSH practices. Objective: This study was carried out with an objective to assess baseline WaSH practices among PLHA and to find out if any association between nutritional status and WaSH parameters so as to identify the shortcomings and highlight the importance of WaSH practices among PLHAs and give suitable recommendations to program managers. Methodology: A cross-sectional study was carried out among PLHA registered in ART centres of western Maharashtra. A sample size of 378 consented to be part of the study were included in the study, by means of systematic random sampling. Data were collected by means of pretested questionnaire prepared from guidelines and previous studies. Institutional ethical clearance was obtained and informed consent was taken from study participants before data collection. Strict confidentiality was maintained throughout the study period. Results: When asked about the water processing method, 76% of them told they do not treat/process the water supplied to them. In contrast, only a few PLHA told they would filter (17%), boil (7%), and use aquaguard (1%). The majority (67%) had their latrines, and while 29% of them were using public latrines and its hygienic sanitation was a concern for them and 4% were still practising open-air defecation in rural areas. Almost all study participants were handwashing after toilet use and handwashing before consuming food. The majority (79%) told they used soap and water, while 20% were using only water, soil and water (1%) and ash and water (1%) which was a concern. Of all the households, 87% cooked their food twice and consumed it thrice a day, while 4% prepared thrice. 10% of them cooked only once and consumed thrice a day; 56% practised consumption of leftover food of the previous night which was a concern. A majority (54%) did not consume street food while remaining said that they used to consume street food. But majority (59%) said they did not follow the habit of checking hygiene or sanitation of hotel where they used to consume food while the remaining 41% did not check the hotel before ordering the food; 50% of them bought packaged milk while 40.9% brought from unpasteurized dairy. On analysis, hygiene and sanitation factors had no statistically significant association with the nutritional status of study participants. Conclusion: WaSH factors act synergistically with other factors to affect the holistic health of PLHA. Information, Education and Communication (IEC) activities (continuous and regular), that focus on improving awareness level on WaSH practices, need to be promoted and integrated into HIV program, including providing basic care packages to PLHA like household water treatment products and soap, etc., Such measures will go a long way in maintaining health of PLHA even during ongoing COVID-19 pandemic scenario.

Variability in Platelet-Rich Plasma Preparations Used in Regenerative Medicine: A Comparative Analysis.

Background: Platelet-rich plasma (PRP) and its derivatives are used in several aesthetic, dental, and musculoskeletal procedures. Their efficacy is primarily due to the release of various growth factors (GF), interleukins, cytokines, and white blood cells. However, the PRP preparation methods are highly variable, and studies lack consistency in reporting complete procedures to prepare PRP and characterize PRP and its derivatives. Also, all the tissue-specific (in vivo and in vitro) interactions and functional properties of the various derivatives/factors of the PRP have not been taken into consideration by any study so far. This creates a potential space for further standardization of the PRP preparation methods and customization of PRP/PRP derivatives targeted at tissue-specific/pathology specific requirements that would enable efficacious and widely acceptable usage of PRP as main therapy, rather than being used as adjuvant therapy. The main objective of our study was to investigate the variability in PRP preparation methods and to analyze their efficacy and reliability. Method: This study considered articles published in the last 5 years, highlighting the variability in their PRP preparation methods and characterization of PRP. Following the PRISMA protocol, we selected 13 articles for the study. The selected articles were assessed using NHLBI quality assessment tool. Results: We noted differences in (1) approaches to producing PRP, (2) extent of characterization of PRP, (3) small scale and large-scale preparation methods, (4) in vitro and in vivo studies. Conclusion: We identified two studies describing the procedures which are simple, reproducible, economical, provide a good yield of platelets, and therefore can be considered methods for further tissue-specific and pathology-specific standardizations of PRP and its derivatives. We recommend further randomized studies to understand the full therapeutic potential of the constituents of PRP and its derivatives.