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Some gene polymorphisms can lead to monogenic diseases, whereas other polymorphisms may confer beneficial traits. A well-characterized example is congenital erythrocytosis-the non-pathogenic hyper-production of red blood cells-that is caused by a truncated erythropoietin receptor. Here we show that Cas9-mediated genome editing in CD34+ human haematopoietic stem and progenitor cells (HSPCs) can recreate the truncated form of the erythropoietin receptor, leading to substantial increases in erythropoietic output. We also show that combining the expression of the cDNA of a truncated erythropoietin receptor with a previously reported genome-editing strategy to fully replace the HBA1 gene with an HBB transgene in HSPCs (to restore normal haemoglobin production in cells with a β-thalassaemia phenotype) gives the edited HSPCs and the healthy red blood cell phenotype a proliferative advantage. Combining knowledge of human genetics with precise genome editing to insert natural human variants into therapeutic cells may facilitate safer and more effective genome-editing therapies for patients with genetic diseases.

Abstract Adaptive Laboratory Evolution (ALE) of microorganisms can improve the efficiency of sustainable industrial processes important to the global economy. However, stochasticity and genetic background effects often lead to suboptimal outcomes during laboratory evolution. Here we report an ALE platform to circumvent these shortcomings through parallelized clonal evolution at an unprecedented scale. Using this platform, we evolved 104 yeast populations in parallel from many strains for eight desired wine fermentation-related traits. Expansions of both ALE replicates and lineage numbers broadened the evolutionary search spectrum leading to improved wine yeasts unencumbered by unwanted side effects. At the genomic level, evolutionary gains in metabolic characteristics often coincided with distinct chromosome amplifications and the emergence of side-effect syndromes that were characteristic of each selection niche. Several high-performing ALE strains exhibited desired wine fermentation kinetics when tested in larger liquid cultures, supporting their suitability for application. More broadly, our high-throughput ALE platform opens opportunities for rapid optimization of microbes which otherwise could take many years to accomplish.

Abstract Deletion of the maternal UBE3A allele causes Angelman syndrome (AS); because paternal UBE3A is epigenetically silenced by a long non-coding antisense (UBE3A-ATS) in neurons, this nearly eliminates UBE3A protein in the brain. Reactivating paternal UBE3A holds promise for treating AS. We previously showed topoisomerase inhibitors can reactivate paternal UBE3A, but their therapeutic challenges prompted our search for small molecule unsilencers with a different mechanism of action. Here, we found that (S)-PHA533533 acts through a novel mechanism to significantly increase paternal Ube3a mRNA and UBE3A protein levels while downregulating Ube3a-ATS in primary neurons derived from AS model mice. Furthermore, peripheral delivery of (S)-PHA533533 in AS model mice induces widespread neuronal UBE3A expression. Finally, we show that (S)-PHA533533 unsilences paternal UBE3A in AS patient-derived neurons, highlighting its translational potential. Our findings provide a lead for developing a small molecule treatment for AS that could be safe, non-invasively delivered, and capable of brain-wide unsilencing of paternal UBE3A.

Introduction: Chitosan nanoparticles (CSN) are widely used for endodontic disinfection; however, their combination with other intracanal medicaments is least studied; therefore, this study aims to evaluate the effect of various intracanal medicaments incorporated with 0.2% CSN on root dentin microhardness and fracture resistance. Materials and Methods: Fifty extracted human single-rooted premolars were cleaned, decoronated, and standardized to 13 mm root length. After access opening and working length determination, specimens were instrumented with ProTaper rotary files till apical size F3 and alternatively irrigated with 5 ml of 2.5% sodium hypochlorite and normal saline (NS). Final irrigation was done with 5 ml of 1% phytic acid. The specimens were allocated randomly into five groups (n = 10), followed by placement of intracanal medicament for 1 week as follows. Group 1 – 0.2% CSN, Group 2 – Calcium hydroxide (Ca[OH] 2) + 0.2% CSN, Group 3 – Triple antibiotic paste (TAP) + 0.2% CSN, Group 4 – Propolis + 0.2% CSN, and Group 5 – Chlorhexidine gluconate (CHX) + 0.2% CSN. The specimens were then rinsed with NS and 3 mm coronal portion of each specimen was horizontally sectioned to create dentin blocks that were further split vertically and tested for Vickers microhardness (n = 10). The remaining root segment of each specimen was then vertically embedded in an acrylic block and evaluated for fracture resistance (n = 10) under a universal testing machine. The data acquired were statistically assessed using one-way ANOVA and posthoc Tukey tests (P < 0.05). Results: Propolis incorporated with 0.2% CSN showed the highest microhardness and fracture resistance values followed by CHX + 0.2% CSN, TAP + 0.2% CSN, Ca(OH)2 + 0.2% CSN, and least in 0.2% CSN alone. Conclusion: Intracanal medicaments incorporated with 0.2% CSN exhibited a synergetic effect on microhardness and fracture resistance of endodontically treated teeth; however, propolis with 0.2% CSN exhibited higher microhardness and fracture resistance of root dentin.

Abstract Cheese fermentation and flavour formation are the result of complex biochemical reactions driven by the activity of multiple microorganisms. Here, we studied the roles of microbial interactions in flavour formation in a year-long Cheddar cheese making process, using a commercial starter culture containing Streptococcus thermophilus and Lactococcus strains. By using an experimental strategy whereby certain strains were left out from the starter culture, we show that S. thermophilus has a crucial role in boosting Lactococcus growth and shaping flavour compound profile. Controlled milk fermentations with systematic exclusion of single Lactococcus strains, combined with genomics, genome-scale metabolic modelling, and metatranscriptomics, indicated that S. thermophilus proteolytic activity relieves nitrogen limitation for Lactococcus and boosts de novo nucleotide biosynthesis. While S. thermophilus had large contribution to the flavour profile, Lactococcus cremoris also played a role by limiting diacetyl and acetoin formation, which otherwise results in an off-flavour when in excess. This off-flavour control could be attributed to the metabolic re-routing of citrate by L. cremoris from diacetyl and acetoin towards α-ketoglutarate. Further, closely related Lactococcus lactis strains exhibited different interaction patterns with S. thermophilus, highlighting the significance of strain specificity in cheese making. Our results highlight the crucial roles of competitive and cooperative microbial interactions in shaping cheese flavour profile.

Custard apple (Annona squamosa L.) is commercially grown for its sweetness, delicacy, and nutraceutical benefits across the world. Fruit must be harvested from trees before the climacteric stage begins for stabilizing the maturation process and perishability, which otherwise restricts longer shelf–life. Therefore, maturity–based grading or classifying custard apple fruit in accordance with their ripening is required for commercial marketing and industrial storage purposes. This is a crucial task that agriculturists and the food processing industries must have to perform manually, which is ineffective and susceptible to human errors. In this research, we have proposed an image processing–based maturity stages detection prototype device for fruit classification of custard apple (Cv. Balanagar) based on changes in their physical, chemical and imaging features (skin color of fruit areoles). The newly developed handheld device comprises of a Raspberry Pi board, camera module, LCD touch screen, and image processing algorithms to identify five essential maturity stages i.e. 0 %, 25 %, 50 %, 75 % and 100 % areoles opening of custard apple. The images were pre–processed and segmented, after which imaging aspects of custard apple fruit including relative R, G and B components were extracted and their relationship with chemical properties such as TSS was established. For classification of captured images, K–means clustering and SVM algorithms were performed on Raspberry Pi Platform with model training and classifier codes in Python (3.9.6) to get desired results. The statistical analysis revealed a substantial difference between the maturity stages in terms of R, G and TSS. Overall, 100 % accuracy was accomplished with respect to the grading of custard apple fruit at varied maturity. This could be a low–cost device useful for farm–level custard apple fruit classification, minimising economic losses caused by faulty supply entering the marketing chain. With slight modifications in algorithms, this device could also be used to assess the maturity of other fruit crops. It could be also an alternative to several expensive approaches being used for automating the maturity grading process that is not feasible, particularly for farmers working in field conditions.

Background: Bladder cancer is the sixth most common malignancy in the United States (US). Despite its high prevalence and the significant potential benefits of early detection, no reliable, cost-effective screening algorithm exists for asymptomatic patients at risk. Nonetheless, reports of incidentally identified early bladder cancer on CT/MRI scans performed for other indications are emerging in the literature. This represents a new opportunity for early detection, with over 80 million CT scans performed in the US yearly, 40% of which are abdominopelvic CTs. This investigation aims to define the imaging features of early bladder cancer, with the mission of facilitating early diagnosis. Methods: Following IRB approval with a waiver of informed consent, a retrospective review was performed, identifying 624 patients with non-muscle-invasive bladder cancer diagnosed at Johns Hopkins Hospital between 2000 and 2019. Of these patients, 99 patients underwent pelvic CT within the 5 years preceding pathologic diagnosis. These imaging studies were reviewed retrospectively to evaluate for the presence and features of any focal bladder wall abnormality. Results: Median age at the time of pathologic diagnosis was 70 years (range: 51–88 years), and 82% (81/99) of patients were male. A total of 226 CT studies were reviewed. The number of studies per patient ranged from 1 to 33. Median time interval between all available imaging and pathologic diagnosis was 14 months. A total of 62% (141/226) of the scans reviewed were performed for indications other than suspected urinary tract cancer (UTC). A bladder wall mass was visualized in 67% (66/99) of patients and on 35% (78/226) of scans performed before diagnosis. The majority (84%, 67/80) of masses were intraluminal. Mean transverse long- and short-axis measurements were 24 mm and 17 mm, respectively, with long dimension measurements ranging between 5 and 59 mm. Conclusions: Early bladder cancer was visualized on CT preceding pathologic diagnosis in more than 2/3 of patients, and the majority of scans were performed for indications other than suspected urinary tract cancer/UTC symptoms. These results suggest that cross-sectional imaging performed for other indications can serve as a resource for opportunistic bladder cancer screening, particularly in high-risk patients.

Abstract Age-associated B cells (ABC) accumulate with age and in individuals with different immunological disorders, including cancer patients treated with immune checkpoint blockade and those with inborn errors of immunity. Here, we investigate whether ABCs from different conditions are similar and how they impact the longitudinal level of the COVID-19 vaccine response. Single-cell RNA sequencing indicates that ABCs with distinct aetiologies have common transcriptional profiles and can be categorised according to their expression of immune genes, such as the autoimmune regulator (AIRE). Furthermore, higher baseline ABC frequency correlates with decreased levels of antigen-specific memory B cells and reduced neutralising capacity against SARS-CoV-2. ABCs express high levels of the inhibitory FcγRIIB receptor and are distinctive in their ability to bind immune complexes, which could contribute to diminish vaccine responses either directly, or indirectly via enhanced clearance of immune complexed-antigen. Expansion of ABCs may, therefore, serve as a biomarker identifying individuals at risk of suboptimal responses to vaccination.

Tomato spotted wilt (TSW) disease caused by tomato spotted wilt orthotospovirus (TSWV, Orthotospovirus tomatomaculae) poses a significant threat to specialty and staple crops worldwide by causing over a billion dollars in crop losses annually. Current strategies for TSWV diagnosis heavily rely on nucleic acid or protein-based techniques which require significant technical expertise, and are invasive, time-consuming, and expensive, thereby catalyzing the search for better alternatives. In this study, we explored the potential of Raman spectroscopy (RS) in early detection of TSW in a non-invasive and non-destructive manner. Specifically, we investigated whether RS could be used to detect strain specific TSW symptoms associated with four TSWV strains infecting three differentially resistant tomato cultivars. In the acquired spectra, we observed notable reductions in the intensity of vibrational peaks associated with carotenoids. Using high-performance liquid chromatography (HPLC), we confirmed that TSWV caused a substantial decrease in the concentration of lutein that was detected by RS. Finally, we demonstrated that Partial Least Squares-Discriminant Analysis (PLS-DA) could be used to differentiate strain-specific TSW symptoms across all tested cultivars. These results demonstrate that RS can be a promising solution for early diagnosis of TSW, enabling timely disease intervention and thereby mitigating crop losses inflicted by TSWV.

The β-lactamase enzyme is a leading cause of drug resistance developed by bacteria. The combination of β-lactam (BL) antibiotics with an appropriate β-lactamase inhibitor (BLI) is a fundamental approach used to circumvent the problem of β-lactamase mediated resistance. In the present study, non-repeated eight clinical E. coli strains were used in the study. Prior to the screening of synthesized compounds, E. coli strains were phenotypically and genotypically characterized on the basis of MIC and PCR amplification respectively. All newly synthesized thirty-five 1, 2, 3-triazole benzoate derivatives were screened against extended-spectrumtrum β-lactamase class A, class B and class C producing clinical E. coli strains. A zone of inhibition was observed against a class C ESBL trait-bearing isolate with a combination disc containing PB 4 and Cefoxitin indicated the inhibitory activity of PB 4. Similarly, inhibitions by PB 6, PB 7, PB 8, PB 9, PB 15, PB 30 and Cefotaxime against class-A ESBL trait-bearing isolates are indicative of class A ESBL inhibitor capacity of these compounds. In conclusion, cefoxitin in combination with PB 4 and cefotaxime in combination PB 6, PB 7, PB 8, PB 9, PB 15, and PB 30 overcomes the class C and class A β-lactamase enzymatic activity respectively in E. coli strains.

Citrus yellow-vein disease (CYVD) was first reported in California in 1957. We now report that CYVD is associated with a virus-like agent, provisionally named citrus yellow-vein associated virus (CYVaV). The CYVaV RNA genome has 2,692 nucleotides and codes for two discernable open reading frames (ORFs). ORF1 encodes a protein of 190 amino acid (aa) whereas ORF2 is presumably generated by a -1 ribosomal frameshifting event just upstream of the ORF1 termination signal. The frameshift product (717 aa) encodes the RNA-dependent RNA polymerase (RdRp). Phylogenetic analyses suggest that CYVaV is closely related to unclassified virus-like RNAs in the family Tombusviridae. Bio-indexing and RNA-seq experiments indicate that CYVaV can induce yellow vein symptoms independently of known citrus viruses or viroids.

Tomato spotted wilt orthotospovirus (TSWV) is one of the most successful pandemic agricultural pathogens transmitted by several species of thrips in a persistent propagative manner. Current management strategies for TSWV heavily rely on growing single-gene resistant cultivars of tomato (“Sw-5b” gene) and pepper (“Tsw” gene) deployed worldwide. However, the emergence of resistance-breaking strains (RB) in recent years has compounded the threat of TSWV to agricultural production worldwide. Despite this, an extensive study on the thrips transmission biology of RB strains is currently lacking. It is also unclear whether mutualistic TSWV-thrips interactions vary across different novel strains with disparate geographical origins. To address both critical questions, we studied whether and how four novel RB strains of TSWV (two sympatric and two allopatric), along with a non-RB strain, impact western flower thrips (WFT) fitness and whether this leads to differences in TSWV incidence, symptom severity (virulence), and virus accumulation in two differentially resistant tomato cultivars. Our findings show that all RB strains increased WFT fitness by prolonging the adult period and increasing fecundity compared to non-RB and non-viruliferous controls, regardless of the geographical origin of strains or the TSWV titers in individual thrips, which were substantially low in allopatric strains. TSWV accumulation in thrips varied at different developmental stages and was unrelated to the infected tissues from which thrips acquired the virus. However, it was significantly positively correlated to that in WFT-inoculated susceptible plants, but not the resistant ones. The TSW incidences were high in tomato plants infected with all RB strains, ranging from 80% to 90% and 100% in resistant and susceptible plants, respectively. However, TSW incidence in the non-RB-infected susceptible tomato plants was 80%. Our findings provide new insights into how novel strains of TSWV, by selectively offering substantial fitness benefits to vectors, modulate transmission and gain a potential epidemiological advantage over non-RB strains. This study presents the first direct evidence of how vector-imposed selection pressure, besides the one imposed by resistant cultivars, may contribute to the worldwide emergence of RB strains.

Abstract Multi‐omics analyses are used in microbiome studies to understand molecular changes in microbial communities exposed to different conditions. However, it is not always clear how much each omics data type contributes to our understanding and whether they are concordant with each other. Here, we map the molecular response of a synthetic community of 32 human gut bacteria to three non‐antibiotic drugs by using five omics layers (16S rRNA gene profiling, metagenomics, metatranscriptomics, metaproteomics and metabolomics). We find that all the omics methods with species resolution are highly consistent in estimating relative species abundances. Furthermore, different omics methods complement each other for capturing functional changes. For example, while nearly all the omics data types captured that the antipsychotic drug chlorpromazine selectively inhibits Bacteroidota representatives in the community, the metatranscriptome and metaproteome suggested that the drug induces stress responses related to protein quality control. Metabolomics revealed a decrease in oligosaccharide uptake, likely caused by Bacteroidota depletion. Our study highlights how multi‐omics datasets can be utilized to reveal complex molecular responses to external perturbations in microbial communities.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Aim: The aim of the study was to evaluate the antimicrobial activity of 1% phytic acid and ethylenediaminetetraacetic acid (EDTA) incorporated with 0.2% of chitosan nanoparticles (Csnps) against Enterococcus faecalis (E. faecalis) by agar diffusion and biofilm model. Materials and Methods: Antimicrobial efficacy was evaluated in two stages: (i) agar well-diffusion method and (ii) biofilm model. E faecalis (American Type Culture Collection 29212) strain was subcultured and grown on the blood agar plates, followed by placement of test solutions in punched holes (5 mm diameter) and incubated for 24 h at 37°C as follows (n = 10): Group 1: normal saline (control), Group 2: EDTA, Group 3: EDTA +0.2% Csnps, Group 4: 1% phytic acid, and Group 5: 1% phytic acid + 0.2% Csnps followed by measurement of inhibition zone. For biofilm model, standardized dentin blocks were prepared from fifty human-extracted single-rooted premolars and sterilized and contaminated for 2 weeks by transferring 50 μl of bacterial inoculum to sterilized test tubes containing 1 mL of Mueller–Hinton agar broth (MHA) with alternate day replenishment of the broth. The specimens were cleaned and subjected to test solutions as mentioned above. Serial decimal dilutions were prepared, followed by inoculation on MHA plates and incubation for 24 h, accompanied by counting of the colonies per milliliter (colony-forming unit per milliliter). Results were statistically analyzed using one-way analysis of variance and Post hoc Tukey's tests (P < 0.05). Results: Phytic acid incorporated with Csnps exhibited a greater zone of inhibition and least colony count against E. faecalis biofilm compared to all groups. Conclusion: Combination of Csnps and phytic acid exhibits effective antimicrobial activity against E. faecalis.

Background: Amid the COVID-19 period and lockdowns, medical education has witnessed major changes. Shifting from face-to-face offline education to online education in such an important branch of education dealing with human lives and serving mankind is really very challenging. In this distance learning process, student satisfaction is important and prediction of their satisfaction and other important parameters will help in improving the online system of medical education. Objective: To evaluate students' perception about distance learning education based on different domains given in the Distance learning Education Environment survey scale. Materials and Methods: A cross-sectional study was conducted among first-year and final-year MBBS students of the Institute of Medical Sciences, BHU. The total number of students who participated in the study was 123. The Distance Education Learning Environments Survey (DELES) scale which was validated for higher education by Scott L was used. The domain represents instructor support (D1), student interaction and collaboration (D2), personal relevance (D3), authentic learning (D4), active learning (D5), student autonomy (D6), and student satisfaction in distance learning process (D7). Results: The mean and standard deviation for each domain of the DELES were calculated. For instructor support, the mean was 25.9 for first-year students and 21.3 for final-year students. For the domain of authentic learning, the mean score was low, i.e. 15.7, for the first year and 13.3 for the final year. In the learning domain for the question about students satisfaction, the mean score was 19.8 for first-year students and 17.9 for final-year students. Conclusion: The finding of this study indicated that the majority of students were satisfied with instructor's support and personal relevance of the online learning classes, but were unsatisfied with opportunities of student interaction and group discussion, which is essential in medical education.

Nucleic acid-based therapeutics that control gene expression have been steadily progressing towards achieving their full clinical potential throughout the last few decades. Rapid progress has been achieved in RNAi-based therapy by optimizing high specificity and gene silencing efficiency using chemically modified siRNAs. Since 2018, four siRNA drugs – patisiran, givosiran, lumasiran, and inclisiran, were approved by the US FDA, providing a testament to the promise of RNAi therapeutics. Despite these promising results, safe and efficient siRNA delivery at the target site remains a major obstacle for efficient siRNA-based therapeutics. In this review, we have outlined the synergistic effects of emerging dual ribose modifications, including 2’,4’- and 2’,5’-modifications, 5’-E/Z-vinylphosphonate, and northern methanocarbacyclic (NMC) modifications that have contributed to drug-like effects in siRNA. These modifications enhance nuclease stability, prolong gene silencing efficiency, improve thermal stability, and exhibit high tissue accumulation. We also highlight the current progress in siRNA clinical trials. This review will help to understand the potential effects of dual ribose modifications and provides alternative ways to use extensive 2’-modifications in siRNA drugs. Moreover, the minimal number of these dual ribose modifications could be sufficient to achieve the desired therapeutic effect. In future, detailed in vivo studies using these dual ribose modifications could help to improve the therapeutic effects of siRNA. Rational design could further open doors for the rapid progress in siRNA therapeutics.

Abstract Adaptive evolution under controlled laboratory conditions has been highly effective in selecting organisms with beneficial phenotypes such as stress tolerance. The evolution route is particularly attractive when the organisms are either difficult to engineer or the genetic basis of the phenotype is complex. However, many desired traits, like metabolite secretion, have been inaccessible to adaptive selection due to their trade‐off with cell growth. Here, we utilize genome‐scale metabolic models to design nutrient environments for selecting lineages with enhanced metabolite secretion. To overcome the growth‐secretion trade‐off, we identify environments wherein growth becomes correlated with a secondary trait termed tacking trait. The latter is selected to be coupled with the desired trait in the application environment where the trait manifestation is required. Thus, adaptive evolution in the model‐designed selection environment and subsequent return to the application environment is predicted to enhance the desired trait. We experimentally validate this strategy by evolving Saccharomyces cerevisiae for increased secretion of aroma compounds, and confirm the predicted flux‐rerouting using genomic, transcriptomic, and proteomic analyses. Overall, model‐designed selection environments open new opportunities for predictive evolution.

Pathogenic bacteria proliferating inside mammalian host cells need to rapidly adapt to the intracellular environment. How they achieve this and scavenge essential nutrients from the host has been an open question due to the difficulties in distinguishing between bacterial and host metabolites in situ. Here, we capitalized on the inability of mammalian cells to metabolize mannitol to develop a stable isotopic labeling approach to track Salmonella enterica metabolites during intracellular proliferation in host macrophage and epithelial cells. By measuring label incorporation into Salmonella metabolites with liquid chromatography-mass spectrometry (LC-MS), and combining it with metabolic modeling, we identify relevant carbon sources used by Salmonella, uncover routes of their metabolization, and quantify relative reaction rates in central carbon metabolism. Our results underline the importance of the Entner-Doudoroff pathway (EDP) and the phosphoenolpyruvate carboxylase for intracellularly proliferating Salmonella. More broadly, our metabolic labeling strategy opens novel avenues for understanding the metabolism of pathogens inside host cells.

Introduction: Early identification of Left Ventricular Hypertrophy (LVH), a powerful and independent predictor of Cardiovascular Disease (CVD), is a key element for preventing Cardiovascular Events (CVE). High-Normal Blood Pressure (HNBP) was significantly associated with a new-onset Electrocardiogram (ECG) diagnosed LVH. The demographic and personal lifestyle characteristics could be related to HNBP in ECG diagnosed LVH. This relation, if found, can be useful as a factor for early identification of HNBP in relation to ECG diagnosed LVH. Aim: To compare demographic and personal lifestyle characteristics with HNBP in relation to ECG diagnosed LVH in healthy adults. Materials and Methods: The cross-sectional study was conducted in AIIMS Raipur (CG) from April 2021 to March 2022 among 95 healthy adult males between 20 to 39 years with a Blood Pressure (BP) of 130-139/85-89 mm Hg and no anti-hypertensive medications. Permission was obtained from the Ethics committee of AIIMS Raipur (CG). Data was collected from the individuals using health questionnaire, Personal Lifestyle Questionnaire (PLQ), validated anthropometric equipment stature meter for measuring height and weighing machine, a working electronic BP measuring instrument, and ECG. The present study included demographic factors like age, height, weight, Body Mass Index (BMI), educational level, marital status, occupation, and Socio-Economic Status (SES) classified based on Kuppuswamy’s classification. The personal lifestyle characteristics included physical activity, use of substances (alcohol, smoking, tobacco, gutkha), and diet. The unpaired t-test, Chi-square test and regression analysis were applied for the analysis of the collected data. Results: Representation of demographic factors were age (31.9±5.08), height (1.69±0.06), weight (68.3±10.2), BMI (23.8±3.05) in terms of (mean±SD) and SES in terms of n (%) were upper I-7 (7.4), upper middle II-31 (32.6), lower middle III-30 (31.6), upper lower IV-25 (26.3), lower V-2 (2.1). SES classes of modified Kuppuswamy’s SES scale had a significant relationship with LVH {p-value

Sleep is a complex and plastic behavior regulated by multiple brain regions and influenced by numerous internal and external stimuli. Thus, to fully uncover the function(s) of sleep, cellular resolution of sleep-regulating neurons needs to be achieved. Doing so will help to unequivocally assign a role or function to a given neuron or group of neurons in sleep behavior. In the Drosophila brain, neurons projecting to the dorsal fan-shaped body (dFB) have emerged as a key sleep-regulating area. To dissect the contribution of individual dFB neurons to sleep, we undertook an intersectional Split-GAL4 genetic screen focusing on cells contained within the 23E10-GAL4 driver, the most widely used tool to manipulate dFB neurons. In this study, we demonstrate that 23E10-GAL4 expresses in neurons outside the dFB and in the fly equivalent of the spinal cord, the ventral nerve cord (VNC). Furthermore, we show that 2 VNC cholinergic neurons strongly contribute to the sleep-promoting capacity of the 23E10-GAL4 driver under baseline conditions. However, in contrast to other 23E10-GAL4 neurons, silencing these VNC cells does not block sleep homeostasis. Thus, our data demonstrate that the 23E10-GAL4 driver contains at least 2 different types of sleep-regulating neurons controlling distinct aspects of sleep behavior.

Whitefly, Bemisia tabaci Gennadius (B cryptic species), transmits cucurbit leaf crumple virus (CuLCrV) in a persistent fashion. CuLCrV affects several crops such as squash and snap bean in the southeastern United States. CuLCrV is often found as a mixed infection with whitefly transmitted criniviruses, such as cucurbit yellow stunting disorder virus (CYSDV) in hosts such as squash, or as a single infection in hosts such as snap bean. The implications of different host plants (inoculum sources) with varying infection status on CuLCrV transmission/epidemics is not clear. This study conducted a series of whitefly mediated CuLCrV transmission experiments. In the first experiment, three plants species: squash, snap bean, and tobacco were inoculated by whiteflies feeding on field-collected mixed-infected squash plants. In the second experiment, three plant species, namely squash, snap bean, and tobacco with varying infection status (squash infected with CuLCrV and CYSDV and snap bean and tobacco infected with CuLCrV), were used as inoculum sources. In the third experiment, squash plants with differential CuLCrV accumulation levels and infection status (either singly infected with CuLCrV or mixed infected with CuLCrV and CYSDV) were used as inoculum sources. Irrespective of plant species and its infection status, CuLCrV accumulation in whiteflies was dependent upon the CuLCrV accumulation in the inoculum source plants. Furthermore, differential CuLCrV accumulation in whiteflies resulted in differential transmission, CuLCrV accumulation, and disease phenotype in the recipient squash plants. Overall, results demonstrate that whitefly mediated CuLCrV transmission between host plants follows a virus density dependent phenomenon with implications for epidemics.

High school students find reaction stoichiometry and the concept of the mole hard to comprehend. Teachers are well aware of the importance of teaching the mole concept and also the difficulty associated with it. In an attempt to improve the understanding of the mole concept with reduced difficulty, an atomic model kit that provides a hands-on learning experience has been developed and implemented in a class of 40 students. Each model atom consists of spherical neodymium magnets (acting as nucleons) of appropriate number (1 for hydrogen, 12 for carbon, 14 for nitrogen, and 16 for oxygen) enclosed in a hollow spherical Styrofoam casing (acting as an atom) that can be opened when required. These model atoms can be easily weighed using a laboratory balance, to obtain mass ratios of 1:12:14:16. This model kit provides an opportunity to have a hands-on learning experience of abstract concepts such as mass conservation in a balanced chemical reaction, excess and limiting reagents, and the mole and its relation to gram atomic/molecular mass. Herein, we provide a detailed description of the design criteria and the construction of the model atoms. We also provide the activity sheets with the necessary instructions to carry out the hands-on activities and the assessment sheets used for evaluation.

Aim: The study aims to evaluate the antimicrobial properties and push-out bond and compressive strength of mineral trioxide aggregate (MTA) mixed with different ratios of triple antibiotic paste (TAP) against Enterococcus faecalis. Methodology: Antimicrobial effect was evaluated using agar well diffusion method. Materials were divided into three groups. Group 1: MTA, Group 2: MTA + TAP (2:1 ratio), and Group 3: MTA + TAP (1:1 ratio) zone of inhibition were determined after 24 h of incubation at 37°C. To evaluate push-out bond strength, 30 extracted teeth were decoronated to a length of 13 mm, followed by root canal treatment, the root resected at 3 mm from the apex, and root-end cavity was prepared and filled. Cylindrical specimens (n = 10 per group) of size 3 mm × 6 mm were prepared of MTA, MTA plus triple antibiotic 2:1 and 1:1 ratio and tested for compressive strength using Instron Universal testing machine. All the data were statistically analysed using Krusal–Wallis test (P < 0.05). Results: MTA + TAP (1:1 ratio) exhibited highest antimicrobial activity than MTA + TAP (2:1 ratio) and MTA alone. The push-out bond and compressive strength of Group 1 (MTA alone) was better compared to two groups containing MTA in combination with TAP; however, no statistical significant difference was found between the three groups. Conclusion: Combination of TAP with MTA increases the antimicrobial activity against E. faecalis without compromising much of the compressive and push-out bond strength, hence can be advocated for root-end filling.

Most of the small-molecule drugs approved for the treatment of cancer over the past 40 years are based on natural compounds. Bacteria provide an extensive reservoir for the development of further anti-cancer therapeutics to meet the challenges posed by the diversity of these malignant diseases. While identifying cytotoxic compounds is often easy, achieving selective targeting of cancer cells is challenging. Here we describe a novel experimental approach (the Pioneer platform) for the identification and development of 'pioneering' bacterial variants that either show or are conduced to exhibit selective contact-independent anti-cancer cytotoxic activities. We engineered human cancer cells to secrete Colicin M that repress the growth of the bacterium Escherichia coli, while immortalised non-transformed cells were engineered to express Chloramphenicol Acetyltransferase capable of relieving the bacteriostatic effect of Chloramphenicol. Through co-culturing of E. coli with these two engineered human cell lines, we show bacterial outgrowth of DH5α E. coli is constrained by the combination of negative and positive selection pressures. This result supports the potential for this approach to screen or adaptively evolve 'pioneering' bacterial variants that can selectively eliminate the cancer cell population. Overall, the Pioneer platform demonstrates potential utility for drug discovery through multi-partner experimental evolution.

Introduction: Remdesivir is a nucleotide analogue prodrug that perturbs viral replication. Remdesivir has been used in various trials previously for the treatment of Coronavirus Disease 2019 (COVID-19). Some clinical trials found that remdesivir had a mortality benefit, while other studies did not. It was also seen that remdesivir shortened the duration of hospital stay among COVID-19 patients in some trials while in other trials remdesivir did not have any influence on the duration of hospital stay. Aim: To study the clinical outcomes of Remdesivir in moderate and severe cases of COVID-19. Materials and Methods: The present retrospective, cohort study was conducted in the Department of General Medicine, SDM Medical College and Hospital, Dharwad, Karnataka, India, from September 2021 to May 2022, in which 400 case records of patients admitted with moderate to severe COVID-19 were studied. Among these 314 patients received remdesivir and 86 patients did not receive remdesivir. Categorical variables, nominal variables were represented as proportions and frequencies and continuous variables were represented as mean±SD. Statistical analysis was done using Chi-square test. Results: A total of 400 COVID-19 patients were included in the study, among which 314 received remdesivir and 86 did not receive remedisivir. Those who received remdesivir had a mortality rate of 13.69% as compared to 11.63% among those who did not receive remdesivir (p-value=0.6170). In the remdesivir group, 36.62% had a hospital stay of >11 days, while it was 30.23% among the non remedisivir group (p-value=0.3060). Conclusion: In this study, remdesivir did not reduce mortality nor did it reduce the duration of hospital stay among moderate to severe COVID-19 patients.

Tomato (Solanum lycopersicum L.) is an important crop that possesses about 35,000 genes. The treatment of plants with elicitors or pathogen attacks causes a cascade of defense reactions. We investigated tomato responses to the BamFXTM solution containing Zn and Cu elicitors and report the results of comparative transcriptome analysis of tomato seeds treated with Zn and Cu elicitors. The seeds were treated with optimum concentrations of Bam-FX solutions and subjected to cold methanolic extraction methods to obtain the secondary metabolites produced within them at different time intervals post-Bam-FX treatment. The metabolite mixture was analyzed using gas chromatography-mass spectrometry (GCMS). In transcriptome sequencing, GO and KEGG analyses revealed that the majority of the DEGs in BamFx-treated tomato was associated with primary and secondary metabolism, plant hormone signal transduction, TF regulation, transport, and responses to stimuli.The secondary metabolites found in the BamFX treated tomato seedlings – Esters of Fumaric acid, Succinic acid etc. The transcript levels of most auxin transporter-encoding genes changed significantly in the BamFX-treated seedlings (e.g., Solyc01g007010.3, a RING-type E3 ubiquitin transferase). The gene Solyc07g061720.3 for Gibberellin 2-oxidase and the Phorbol-ester/DAG-type domain-containing protein (Solyc02g068680.1) associated with the intracellular signaling genes were found upregulated in the BamFx-treated seeds. The time-dependent effect of the BamFX (1:500 for 60 min) was found to be regulating Abscisic acid signaling pathway genes (Solyc09g015380.1). This study identified many candidate genes for future functional analyses and laid a theoretical foundation for an improved understanding of the molecular mechanisms involved in the BamFx treatment of tomatoes to improve stress resistance.