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Abstract Liquid-like protein condensates are ubiquitous in cellular system and are increasingly recognized for their roles in physiological processes. Condensed phase harbors distinctive chemical microenvironment, markedly different than dilute aqueous phase. Herein, we demonstrate chemoselective modification pattern of nucleophilic canonical amino acid sidechains (namely – cysteine, tyrosine and lysine) of the protein towards 4-chloro-7-nitrobenzofurazan in the dilute and condensed phase. We also delineate how the effect of nucleotides and their in situ enzymatic dissociation temporally modulate the protein condensate’s pH and the protein’s corresponding chemoselective modification. We have shown that the pH of the condensate decreases in the presence of nucleoside triphosphate, whereas it increases in the presence of nucleoside monophosphates or phosphate ion. For instance, we find lysine-specific modification gets inhibited in the presence of adenosine triphosphate (ATP), but significantly enhanced in the presence of monophosphates. This feature enables us to gain temporal control over dynamic change in protein functionalization via enzymatic ATP hydrolysis. Overall, this work substantiates the alteration in pH-responsiveness of Brønsted basicity of a protein’s ε-amine in the condensed phase. Furthermore, this environment sensitivity in chemoselective protein functionalization in condensed phase will be important in adaptable protein engineering to the chemical biology of protein phase separation.

Abstract Background For centuries, plants have been used in the folk medicine of various cultures for their healing properties. It is amazing how nature has provided us with such powerful remedies. Oxalis corniculata (O. corniculata) has always been used traditionally for its medicinal attributes. In Asia, this herbaceous plant is used for treating many gastrointestinal disorders, such as diarrhea and dysentery. In African folk medicine, on the other hand, this herb is used for respiratory diseases, skin diseases, and fever. Main body Scientific research has revealed numerous pharmacological potentials of this plant, including antitumor, antimicrobial, antioxidant, anti-inflammatory, and hepatoprotective properties. In addition, studies have indicated that the extract of this plant protects against oxidative stress, inflammation, and various diseases, including cancer and diabetes. Phytochemical analysis of O. corniculata revealed various relevant compounds, including phenolic acids, flavonoids, and alkaloids, which are responsible for its therapeutic properties. Conclusion The various constituents of this plant have significant ethnomedicinal potential. The plant is a possible source of extracts and chemical compounds with pharmacological activity. In the future, O. corniculata could have an effective role in the development of modern drugs. The objective of this review is to summarize the current knowledge on the medicinal potential of O. corniculata, including its bioactive compounds, mechanisms of action, and therapeutic applications. Graphical abstract

Abstract Fourth order extended Fisher Kolmogorov reaction diffusion equation has been solved numerically using a hybrid technique. The temporal direction has been discretized using Crank Nicolson technique. The space direction has been split into second order equation using twice continuously differentiable function. The space splitting results into a system of equations with linear heat equation and non linear reaction diffusion equation. Quintic Hermite interpolating polynomials have been implemented to discretize the space direction which gives a system of collocation equations to be solved numerically. The hybrid technique ensures the fourth order convergence in space and second order in time direction. Unconditional stability has been obtained by plotting the eigen values of the matrix of iterations. Travelling wave behaviour of dependent variable has been obtained and the computed numerical values are shown by surfaces and curves for analyzing the behaviour of the numerical solution in both space and time directions.

Extreme weather events and global climate change have exacerbated the problem of evaporation rates. Thus, accurately predicting soil moisture evaporation rates affecting soil cracking becomes crucial. However, less is known about how novel feature engineering techniques and machine-learning predictions may account for estimating the soil moisture evaporation rate. This research focuses on predicting the evaporation rate of soil using machine learning (ML) models. The dataset comprised twenty-one ground-based parameters, including temperature, humidity, and soil-related features, used as features to predict evaporation potential. To tackle the high number of features and potential uncorrelated features, a novel guided backpropagation-based feature selection technique was developed to rank the most relevant features. The top-10 features, highly correlated with evaporation rate, were selected for ML model input, alongside the top-5 and all features. Several ML models, including multiple regression (MR), K-nearest neighbor (KNN), multilayer perceptron (MLP), sequential minimal optimization regression (SMOreg), random forest (RF), and a novel K-Nearest Oracles (KNORA) ensemble, were constructed for the purpose of forecasting the evaporation rate. The average error of these models was assessed using the root mean squared error (RMSE). Experimental results showed that the KNORA ensemble model performed the best, achieving a 7.54 mg/h RMSE in testing with the top-10 features. MLP was followed closely by a 25.1 mg/h RMSE in the same testing. An empirical model using all features showed a higher RMSE of 1319.1 mg/h, indicating the superiority of the ML models for accurate evaporation rate predictions. We highlight the implications of our results for climate-induced soil cracking in the real world.

Having an optimum capital structure is crucial as it leads the company to its better operating performance. This paper examines the key determinants affecting the financing decision of start-up firms with reference to the Delhi NCR region in India by using panel data regression models. The hypotheses are formed based on theories of capital structure and existing literature. The financial information data of a final sample of 29 manufacturing start-up firms are taken into consideration for empirical analysis. The results of this research revealed that firm size, growth opportunities, profitability and liquidity are key factors significantly affecting the capital structure decision of start-up firms in India. The relationship found for firm size, profitability and liquidity supported the hypothesis of pecking order theory while growth variable results supported the hypothesis of trade-off theory. Therefore, the pecking order theory is found to be more applicable here in startup firms in India.

Abstract E2F1 induces hundreds of protein-coding genes influencing diverse signaling pathways but much less is known about its non-coding RNA targets. For identifying E2F1-dependent oncogenic long non-coding RNAs (lncRNAs), we carried out genome-wide transcriptome analysis and discovered an lncRNA, EMSLR, which is induced both in lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). EMSLR depletion blocks the cells in G1 phase and inhibits the clonogenic ability indicating that it is essential for the tumor-related phenotypes. We discovered that EMSLR represses the promoter activity of another lncRNA, LncPRESS1, which is located 6.9 kb upstream of EMSLR and they display an inverse expression pattern in lung cancer cell lines. Depletion of C-MYC results in downregulation of EMSLR and simultaneous upregulation of EMSLR target LncPRESS1, exemplifying how C-MYC and E2F1 signal transduction pathways control the network of lncRNA genes to modulate cell proliferation and differentiation.

Fluorescent dyes offer a useful method for the measurement of intracellular lipids. They are inexpensive and require simple optical measurement instrumentation, whilst simultaneously providing high throughput application. Nile Red is a hydrophobic, metachromatic dye which has been widely used for detection of intracellular lipids. However, Nile Red fluorescence depends on its concentration, microenvironment polarity, incubation time and, therefore, requires strain specific optimization. Hence, neutral lipids in Chlorella emersonii and Pseudokirchneriella subcapitata cannot be quantified using existing Nile Red methods developed for other microalgae strains and, therefore an optimised procedure for these strains is required. In this method development, the optimal excitation and emission wavelengths were selected based on the solvent used for Nile Red dissolution. The effect of Nile Red concentration, microalgae cell concentration, incubation time on fluorescence intensity was explored and optimised. Quintuplet assay repeats were executed for increased assay robustness for two microalgae strains, Chlorella emersonii and Pseudokirchneriella subcapitata, with protocol reliability confirmed by fluorescence microscopy. In brief, 20% (v/v) DMSO containing 10μg/ml and 5μg/ml Nile red was found to be ideal concentration for neutral lipid estimation in Chlorella emersonii and Pseudokirchneriella subcapitata respectively when an incubation time of 60mins and 40mins at 40°C was used. This optimised Nile Red protocol is a robust, simple and cost-effective method for neutral lipid quantification in Chlorella emersonii and Pseudokirchneriella subcapitata.

An experiment was conducted at RARI, Durgapura, SKNAU, Jobner during kharif 2020 to analyze the genetic diversity of the 12 Macrophomina phaseolina isolates by using simple sequence repeat (SSR) markers. The SSR primers were tested for amplification of genomic DNA of Macrophomia phaseolina isolates. Total five primers of MB series were screened against twelve Macrophomia phaseolina isolates. All primes amplified scorable banding pattern. Level of polymorphism for SSR analysis ranged between 60–85.7%. The similarity matrix indicated that 12 isolates exhibited 42.82–100% similarity coefficient. The SSR analysis shows high molecular variability among the isolates grouped into five major clusters A, B, C, D and E. In the similarity matrix Mp-DPR was found to have higher value of similarity coefficient (1), whereas Mp-JSM was found to have lower value if similarity coefficient (0.42).

In present study, 32 diverse genotypes of muskmelon (Cucumis melo L.) collected from different parts of the country were evaluated to study the correlation, path analysis and divergence for yield and quality characters using 18 morphological traits. In cluster analysis, genotypes were grouped into eight distinct clusters with a maximum of 13 genotypes in cluster I. The highest inter-cluster distance (355.14) was obtained between cluster V and VIII. Among the traits studied, shelf-life (54.23%), total soluble solids (27.82%) and fruit weight(7.26%) contributed a major share in the divergence of the genotypes. Four genotypes, viz. VRMM-315, VRMM- 12, Durgapura Madhu, and Jodhpur local accommodated into a single cluster, probably share a genetic similarity. The genotypes of cluster V (Punjab Sunehri, VRMM-205, VRMM-194) and cluster VIII (Jodhpur Local) being most divergent from others may serve as potential parents for breeding programmes.

The high cost of soluble enzymes can limit their use for commercial and industrial purposes. Immobilization can enhance enzyme reusability, thereby reducing product isolation costs and overcoming this economic barrier. In the current study, two novel, purified lipases from Pseudomonas sp. (Pseudomonas reinekei and Pseudomonas brenneri) were entrapped in a calcium alginate matrix, with the aim of simultaneously enhancing enzyme reusability and stability. Following entrapment, the retained activity of the enzyme-alginate composite was verified by an enzymatic hydrolysis reaction of a p-nitrophenol palmitate substrate. The effect of the enzyme-alginate entrapment against various physiochemical parameters such as pH, temperature, metal ions, and solvents were subsequently examined. The entrapment was found to have minimal beneficial stability gains. However, enhanced enzyme reusability (up to 3 cycles) and stability (up to 18 days at 4°C) of the calcium alginate entrapped lipase, as indicated by residual hydrolysis of p-nitrophenol palmitate, was observed, suggesting potential roles for calcium alginate entrapped lipases in cost efficient enzyme catalysis. HIGHLIGHTS •Two novel lipases have been entrapped in calcium alginate for the first time. •A statistically enhanced stability in 1M EDTA was observed following entrapment. •The novel entrapped lipases display excellent storage stability and reusability.

Opium poppy is one of the most important medicinal plant species cultivated in several regions of the world for pharmaceutical and food purposes. Although it has been bred and used in many countries for centuries, taxonomical studies on the intraspecific variability are still not numerous. In the course of this study, the genetic diversity of 58 opium poppy accessions were assessed with SSR markers. Out of the 25 tested SSR markers, only 19 showed successful amplification and seven were found to reveal polymorphism. These seven markers produced 2–11 alleles within the size range of 144–280 bp. PIC value varied between 0.284 and 0.767, expected heterozygosity was between 0.136 and 1, and observed heterozygosity was between 0.344 and 0.794. Altogether, a low genetic diversity was found, and several accessions have been characterized with the same SSR allele profile. The clustering in the PCoA co-ordinate system indicates four groups, which is confirmed by the STRUCTURE analysis as well and reflects the known relationships of hybrid cultivars in several cases. The landrace taxa clearly deviate from the majority of cultivars selected for industrial purposes. The applied SSR markers were not able to distinguish the genotypes according to their geographic origin or seed color. However, in several cases, the grouping in the PCoA system reflected a diversification connected to alkaloid content and petal color of the accessions. Our results demonstrate the importance of further molecular marker studies focusing on genotypes with different alkaloid content and useful agronomical traits in order to establish more effective breeding.

Abstract Background Although net ultrafiltration (UFNET) is frequently used for treatment of fluid overload in critically ill patients with acute kidney injury, the optimal intensity of UFNET is unclear. Among critically ill patients with fluid overload receiving renal replacement therapy (RRT), we examined the association between UFNET intensity and risk-adjusted 1-year mortality. Methods We selected patients with fluid overload ≥ 5% of body weight prior to initiation of RRT from a large academic medical center ICU dataset. UFNET intensity was calculated as the net volume of fluid ultrafiltered per day from initiation of either continuous or intermittent RRT until the end of ICU stay adjusted for patient hospital admission body weight. We stratified UFNET as low (≤ 20 ml/kg/day), moderate (> 20 to ≤ 25 ml/kg/day) or high (> 25 ml/kg/day) intensity. We adjusted for age, sex, body mass index, race, surgery, baseline estimated glomerular filtration rate, oliguria, first RRT modality, pre-RRT fluid balance, duration of RRT, time to RRT initiation from ICU admission, APACHE III score, mechanical ventilation use, suspected sepsis, mean arterial pressure on day 1 of RRT, cumulative fluid balance during RRT and cumulative vasopressor dose during RRT. We fitted logistic regression for 1-year mortality, Gray’s survival model and propensity matching to account for indication bias. Results Of 1075 patients, the distribution of high, moderate and low-intensity UFNET groups was 40.4%, 15.2% and 44.2% and 1-year mortality was 59.4% vs 60.2% vs 69.7%, respectively (p = 0.003). Using logistic regression, high-intensity compared with low-intensity UFNET was associated with lower mortality (adjusted odds ratio 0.61, 95% CI 0.41–0.93, p = 0.02). Using Gray’s model, high UFNET was associated with decreased mortality up to 39 days after ICU admission (adjusted hazard ratio range 0.50–0.73). After combining low and moderate-intensity UFNET groups (n = 258) and propensity matching with the high-intensity group (n = 258), UFNET intensity > 25 ml/kg/day compared with ≤ 25 ml/kg/day was associated with lower mortality (57% vs 67.8%, p = 0.01). Findings were robust to several sensitivity analyses. Conclusions Among critically ill patients with ≥ 5% fluid overload and receiving RRT, UFNET intensity > 25 ml/kg/day compared with ≤ 20 ml/kg/day was associated with lower 1-year risk-adjusted mortality. Whether tolerating intensive UFNET is just a marker for recovery or a mediator requires further research.

Abstract Background Most studies of acute kidney injury (AKI) have focused on older adults, and little is known about AKI in young adults (16–25 years) that are cared for in an adult intensive care unit (ICU). We analyzed data from a large single-center ICU database and defined AKI using the Kidney Disease Improving Global Outcomes criteria. We stratified patients 16–55 years of age into four age groups for comparison and used multivariable logistic regression to identify associations of potential susceptibilities and exposures with AKI and mortality. Results AKI developed in 52.6% (n = 8270) of the entire cohort and in 39.8% of the young adult age group (16–25 years). The AUCs for the age categories were similar at 0.754, 0.769, 0.772, and 0.770 for the 16–25-, 26–35-, 36–45-, and 45–55-year age groups, respectively. For the youngest age group, diabetes (OR 1.89; 95% CI 1.09–3.29), surgical reason for admission (OR 1.79; 95% CI 1.44–2.23), severity of illness (OR 1.02; 95% CI 1.02–1.03), hypotension (OR 1.13; 95% CI 1.04–1.24), and certain medications (vancomycin and calcineurin inhibitors) were all independently associated with AKI. AKI was a significant predictor for longer length of stay, ICU mortality, and mortality after discharge. Conclusions AKI is a common event for young adults admitted to an adult tertiary care center ICU with an associated increased length of stay and risk of mortality. Potentially modifiable risk factors for AKI including medications were identified for all stratified age groups.

Typical bioprocess comprises of different unit operations wherein a near optimal environment is required for cells to grow, divide, and synthesize the desired product. However, bioprocess control caters to unique challenges that arise due to non-linearity, variability, and complexity of biotech processes. This article presents a review of modern control strategies employed in bioprocessing. Conventional control strategies (open loop, closed loop) along with modern control schemes such as fuzzy logic, model predictive control, adaptive control and neural network-based control are illustrated, and their effectiveness is highlighted. Furthermore, it is elucidated that bioprocess control is more than just automation, and includes aspects such as system architecture, software applications, hardware, and interfaces, all of which are optimized and compiled as per demand. This needs to be accomplished while keeping process requirement, production cost, market value of product, regulatory constraints, and data acquisition requirements in our purview. This article aims to offer an overview of the current best practices in bioprocess control, monitoring, and automation.

The present investigation was conducted to study the prevalence and molecular characterization of chlamydiae associated with the respiratory infections of equines in the Sub-Himalayan region of India. Equine nasal swab samples (119) from 20 diseased and 99 apparently healthy equines were collected and tested by family Chlamydiaceae specific nested PCR based on ompA gene, besides 89 serum samples for AGPT. The molecular characterization of chlamydial species/strains was done by analyzing variation in VD II region of ompA gene. The chlamydia infections were detected in 48.74% of the nasal swabs (55% in diseased and 47.47% in apparently healthy animals) by nested PCR. Prevalence of two genetically variant strains of Chlamydia abortus and a single strain of Chlamydia psittaci was detected. AGPT showed 4.49% seropositive equines. High prevalence of chlamydiae was found among equines in the Sub-Himalayan region of India in Himachal Pradesh in both healthy and diseased equines.

Lumen-apposing metal stents (LAMS) use in gastrointestinal endoscopy has been on the rise for various indications for the last few years. Currently, LAMS is a well-established treatment for post-pancreatitis peri-pancreatic fluid collections and walled-off necrosis (WON), but it is still not a standard of care in the treatment of post-surgical fluid collections (PSFC). Most of the earlier studies for treatment of PSFC utilized double pigtail plastic stents (DPS). We present a series of 3 cases where LAMS was successfully used for PSFC drainage. The cases include a patient with perigastric abscess after Whipple’s procedure, a case of peri-pancreatic collection after distal pancreatectomy, and a patient with peri-pancreatic fluid collection after right partial hepatectomy and splenectomy due to lacerations from a motor vehicle accident.

Leafy vegetables contributes to the inflation rate in Ambon City due to low productivity in rainy season. Some vegetables are imported from other islands while importantvegetables such as local petsai (Brassica chinensis L.) and chili (Capsicum annum L.) are cultivated in low nitrogen soil, Entisols. Lack of nitrogen could be overcome by using inorganic fertilizeras well as biofertilzer. The soil can be inoculated with rhizobacteria, such as Azotobacter, to increase the nitrogen uptake and improve the quality of vegetables. This research was conducted to isolate and select Azotobacter from rhizosphere of vegetables and to examine the effect of Azotobacter inoculation on chili-seedling growth and nitrogen uptake by using bioassay method. Azotobacter sp. was isolated in nitrogen-free Ashby’s Media. The bioassay was held in the green house with randomized block design experiment, which examined the combination of isolates and population of Azotobacter sp. on chili. Two best isolates which was selected based on pH, nitrogen content and cell viability were s2a10 (from petsai's rhizosphere) and c2a9 (from chili’s rhizosphere). Bioassay showed that Azotobacter inoculation followed by reduced NPK fertilizer doses had no effect on transplant dry weight and nitrogen uptake. All Azotobacter 8 -1inoculation except 10 CFU mL s2a10 maintain soil nitrogen although Azotobacter population in soil was slightly reduced. This showed that Azotobacter sp. potentially reduce the use of inorganic biofertilizer.

Aim/Purpose: This paper focuses on designing and implementing the rubric for objective JAVA programming assessments. An unsupervised learning approach was used to group learners based on their performance in the results obtained from the rubric, reflecting their learning ability. Background: Students' learning outcomes have been evaluated subjectively using a rubric for years. Subjective assessments are simple to construct yet inconsistent and biased to evaluate. Objective assessments are stable, reliable, and easy to conduct. However, they usually lack rubrics. Methodology: In this study, a Top-Down assessment approach is followed, i.e., a rubric focused on the learning outcome of the subject is designed, and the proficiency of learners is judged by their performance in conducting the task given. A JAVA rubric is proposed based on the learning outcomes like syntactical, logical, conceptual, and advanced JAVA skills. A JAVA objective quiz (with multiple correct options) is prepared based on the rubric criteria, comprising five questions per criterion. The examination was conducted for 209 students (100 from the MCA course and 109 from B.Tech. course). The suggested rubric was used to compute the results. K-means clustering was applied to the results to classify the students according to their learning preferences and abilities. Contribution: This work contributes to the field of rubric designing by creating an objective programming assessment and analyzing the learners' performance using machine learning techniques. It also facilitates a reliable feedback approach offering various possibilities in student learning analytics. Findings: The designed rubric, partial scoring, and cluster analysis of the results help us to provide individual feedback and also, group the students based on their learning skills. Like on average, learners are good at remembering the syntax and concepts, mediocre in logical and critical thinking, and need more practice in code optimization and designing applications. Recommendations for Practitioners: The practical implications of this work include rubric designing for objective assessments and building an informative feedback process. Faculty can use this approach as an alternative assessment measure. They are the strong pillars of e-assessments and virtual learning platforms. Recommendation for Researchers: This research presents a novel approach to rubric-based objective assessments. Thus, it provides a fresh perspective to the researchers promising enough opportunities in the current era of digital education. Impact on Society: In order to accomplish the shared objective of reflective learning, the grading rubric and its accompanying analysis can be utilized by both instructors and students. As an instructional assessment tool, the rubric helps instructors to align their pedagogies with the students' learning levels and assists students in updating their learning paths based on the informative topic-wise scores generated with the help of the rubric. Future Research: The designed rubric in this study can be extended to other programming languages and subjects. Further, an adaptable weighted rubric can be created to execute a flexible and reflective learning process. In addition, outcome-based learning can be achieved by measuring and analyzing student improvements after rubric evaluation.

IntroductionAbout one-third of critically ill patients with acute kidney injury (AKI) develop persistently decreased kidney function, known as acute kidney disease (AKD), which may progress to chronic kidney disease (CKD). Although sepsis is the most common cause of AKI, little is known about sepsis-associated AKD.MethodsUsing data from a large randomized trial including 1341 patients with septic shock, we studied patients with stage 2 or 3 AKI on day 1 of hospitalization. We defined AKD as a persistently reduced glomerular filtration rate for >7 days. In addition to clinical data, we measured several urinary biomarkers (tissue inhibitor of metalloproteinases-2 and insulin-like growth factor-binding protein 7 [TIMP-2∗IGFBP7], neutrophil gelatinase-associated lipocalin [NGAL], kidney injury molecule-1 [KIM-1], liver-type fatty acid binding protein, and type 4 collagen) at 0, 6, and 24 hours, to predict AKD.ResultsOf 598 patients, 119 (19.9%) died within 7 days, 318 (53.2%) had early reversal of AKI within the first 7 days, whereas 161 (26.9%) developed AKD. In patients with early reversal, 45 (14.2%) had relapsed AKI after early reversal, and only about one-third of these recovered. Among patients developing AKD, only 15 (9.3%) recovered renal function prior to discharge. Male sex, African American race, and underlying CKD were more predominant in patients developing AKD. None of the biomarkers tested performed well for prediction of AKD, although NGAL modestly increased the performance of a clinical model.ConclusionsAKD is common in patients with septic shock, especially among African American males and those with underlying CKD. Existing AKI biomarkers have limited utility for predicting AKD but might be useful together with clinical variables. Novel predictive biomarkers for renal recovery are needed.