Quantitative PCR (qPCR) is a highly sensitive, cost effective, and routinely used molecular assay for analyzing the gene expression patterns of specific target genes across tissues, pathological conditions, treatment regimes, and physiological states. However, normalization of expression profiles of target genes using stable reference genes (RGs) is a critical step to ensure the accuracy of relative quantification using qPCR. In this study, we evaluated the stability of fourteen potential existing reference genes (ACTB, BACH1, B2M, GAPDH, HMBS, PGK1, PPIA, PPIB, RPLP0, RPL19, RPS9, RPS15, RPS28, and UXT) in the peripheral blood mononuclear cells (PBMCs) of healthy sheep and goats to determine the most stable RGs. These candidate genes belong to different functional classes and were chosen based on published literature on commonly used RGs in different livestock species. Four different analytical approaches (geNorm, NormFinder, BestKeeper, and ΔCt analysis) as well as RefFinder, an online tool which integrates the geometric means of these four prominent stability algorithms were utilized to determine a comprehensive ranking of the investigated genes. Our data indicates that PPIB, BACH1, ACTB, and PPIA are the most suitable RGs, while RPLP0, GAPDH and RPS15 are the most variable and unsuitable genes for normalization of qPCR data in the PBMCs of sheep and goats. The results of this study provide useful resource for researchers engaged in unravelling the transcriptional landscape of PBMCs of small ruminants for various scientific investigations.

Structural insight eludes on how full-length gelsolin depolymerizes and caps filamentous (F-)actin, while the same entity can nucleate polymerization of G-actins. Analyzing small angle X-ray scattering (SAXS) data, we deciphered assemblies which enable these contrasting processes. Mixing Ca2+-gelsolin with F-actin in high salt F-buffer resulted in depolymerization of ordered F-actin rods to smaller sized species which became monodispersed upon dialysis with low salt G-buffer. These entities were the ternary (GA2) and binary (GA) complexes of gelsolin and actin with radius of gyration and maximum linear dimension of 4.55 and 4.68 nm, and 15 and 16 nm, respectively. Using size exclusion chromatography in-line with SAXS, we confirmed that initially GA and GA2 species are formed as seen upon depolymerization of F-actin followed by dialysis. Interestingly, while GA2 could seed formation of native-like F-actin in both G- and F-buffer, GA failed in G-buffer. Thus, GA2 and GA are the central species formed via depolymerization or towards nucleation. SAXS profile referenced modeling revealed that: 1) in GA, actin is bound to the C-terminal half of gelsolin, and 2) in GA2, second actin binds to the open N-terminal half accompanied by dramatic rearrangements across g1-g2 and g3-g4 linkers.

Background Quantitative real-time PCR (qPCR) is a highly reliable method for validating gene expression data in molecular studies due to its sensitivity, specificity, and efficiency. To ensure accurate qPCR results, it’s essential to normalize the expression data using stable reference genes. Methods This study aimed to identify suitable reference genes for qPCR studies in goats by evaluating 18 candidate reference genes ( ACTB, BACH1, B2M, GAPDH, HMBS, HPRT1, PGK1, PPIA, PPIB, RPLP0, RPL19, RPS9, RPS15, RPS28, SDHA, TBP, UXT , and YWHAZ ) in 10 different caprine tissues (heart, intestine, kidney, liver, lung, muscle, rumen, skin, spleen, and testis). An integrated tool called RefFinder, which incorporates various algorithms like NormFinder, GeNorm, BestKeeper, and ΔCt, was used to assess the stability of expression among these genes. Results After thorough analysis, ACTB, PPIB , and B2M emerged as the most stable reference genes, while RPL19, RPS15 , and RPS9 were found to be the least stable. The suitability of the selected internal control genes was further validated through target gene analysis, confirming their efficacy in ensuring accurate gene expression profiling in goats. Conclusion The study determined that the geometric average of ACTB, PPIB , and B2M creates an appropriate normalization factor for gene expression studies in goat tissues.

BACKGROUND: Osteoarthritis (OA) is a chronic disease in which the cartilage in the joints deteriorates and the bones rub against each other, causing pain, stiffness, and restricted range of motion. This is an age-related condition that affects initially isolated joints or joints on one aspect of the body. The aim of the study is to identify quality of life and self-reported disability in patients with osteoarthritis. MATERIALS AND METHODS: A cross-sectional descriptive study was conducted in Orthopedic O.P.D. of tertiary care hospital. Study was conducted on 150 samples with convenience sampling at orthopedic O.P.D. Data were collected with standardized tools SF-36 with domains physical functioning (PF), role physical (RP), vitality (VT), mental health (MH), role emotional (RE), social functioning (SF), bodily pain (BP) general health (GH), and Western Ontario and McMaster Universities Arthritis Index (WOMAC) questionnaires with domains pain, stiffness, and functional disability. Descriptive statistics and inferential statistics were used for data analysis such as mean, frequency, percentage, standard deviation, and Chi-square test. RESULTS: Out of 150 samples, 103 were females, 114 were Hindu, and 131 were married. Highest mean score in RE domain of SF-36 was 60 with SD 38.43 indicating that patients had low impact on quality of life, whereas lowest mean score was 35.33 with SD 32.67 indicating that patients had severe impact in RP domain. In WOMAC index, patients had highest pain in climbing stairs, stiffness during morning, and functional difficulty during doing heavy domestic work, whereas lowest pain in resting, stiffness in evening, and functional difficulty during lying in bed. CONCLUSION: Patients with OA had poorer quality of life in domains PF, RP, VT, BP, and GH. Patients with osteoarthritis showed highest self-reported disability in terms of pain in climbing stairs, stiffness during morning, and functional difficulties in doing heavy domestic duties.

Normalization of gene expression data using appropriate reference genes is critical to diminish any technical bias in an experiment involving quantitative real-time PCR (qPCR). To the best of our knowledge, this is the first report offering a systematic assessment of 14 potential reference genes (RPLP0, ACTB, RPS28, YWHAZ, SDHA, PPIA, RPS9, RPS15, UXT, GAPDH, B2M, BACH1, HMBS, and PPIB) for the identification of the most stable normalizers for qPCR of target genes in peripheral blood mononuclear cells (PBMCs) of bovines for vector-borne haemoparasitic diseases such as anaplasmosis, babesiosis, theileriosis, and trypanosomiasis. A total of 38 blood samples were collected from healthy as well as diseased cattle and buffaloes representing different haemoparasitic diseases. RNA isolated from the PBMCs was subjected to qPCR for the 14 prospective internal control genes. The comprehensive ranking of the genes was accomplished by the RefFinder tool that integrates the results of three algorithms (geNorm, NormFinder, and BestKeeper) and the comparative CT method. RPS15, B2M, and GAPDH were ranked to be the most stable genes, whereas, PPIA and HMBS emerged to be the least suitable genes. Validation of the selected reference genes by the qPCR analysis of two immunity genes, ISG15 and GPX7 was congruent with the observations of this study. We recommend that a panel of three reference genes including RPS15, B2M, and GAPDH could prove useful in delineating the transcriptional landscape of PBMCs for vector-borne haemoparasitic diseases in bovines.

Bovine anaplasmosis caused by Anaplasma marginale is a tick-borne disease of livestock with widespread prevalence and huge economic implications. In order to get new insights into modulation of host gene expression in response to natural infections of anaplasmosis, this study is the first attempt that compared the transcriptome profiles of peripheral blood mononuclear cells (PBMCs) of A. marginale infected and healthy crossbred cattle. Transcriptome analysis identified shared as well as unique functional pathways in the two groups. Translation and structural constituent of ribosome were the important terms for the genes abundantly expressed in the infected as well as healthy animals. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of the differentially expressed genes revealed that immunity and signal transduction related terms were enriched for the up-regulated genes in the infected animals. The over-represented pathways were cytokine-cytokine receptor interaction and signaling pathways involving chemokines, Interleukin 17 (IL17), Tumour Necrosis Factor (TNF), Nuclear Factor Kappa B (NFKB) etc. Interestingly, many genes previously associated with parasite-borne diseases such as amoebiasis, trypanosomiasis, toxoplasmosis, and leishmaniasis were profusely expressed in the dataset of the diseased animals. High expression was also evident for the genes for acute phase response proteins, anti-microbial peptides and many inflammatory cytokines. Role of cytokines in mediating communication between immune cells was the most conspicuous gene network identified through the Ingenuity Pathway Analysis. This study provides comprehensive information about the crosstalk of genes involved in host defense as well as parasite persistence in the host upon infection with A. marginale.

The molecular changes occurring in the host in response to in vivo Theileria annulata parasitic infection are not well understood. Therefore, the present study investigated the differential expression profiles of peripheral blood mononuclear cells (PBMCs) across Theileria annulata-infected and non-infected crossbred cows. The differential expression profiles from PBMCs of infected and non-infected crossbred cows were generated by RNA sequencing. A marked difference in the expression of genes associated with innate immunity (FTH1, ACTB, ISG15) was observed between the two groups. The over-represented pathways in Theileria annulata-infected cows were associated with the immune system and regulation of the mitotic cycle. Enriched genes and pathways in non-infected animals were associated with the maintenance of chromatin integrity and cell structure. The highly connected genes identified in this study form potential candidates for further investigation into host-parasite interactions in cattle. An improved understanding of the transcriptomic dynamics during theileriosis would lead to underpinning molecular level differences related to the health status of cattle.

Quantitative PCR (qPCR) is a widely-used technique for quantifying the expression of target genes across various tissues, as well as under different pathological and physiological conditions. One of the challenges associated with this method is the need to identify optimal reference genes (RGs) that maintain consistent expression levels under diverse experimental settings, thereby ensuring accurate biological interpretation. In this study, we conducted a thorough analysis of 18 candidate RGs (ACTB, BACH1, B2M, GAPDH, HMBS, HPRT1, PGK1, PPIA, PPIB, RPLP0, RPL19, RPS9, RPS15, RPS28, SDHA, TBP, UXT, and YWHAZ) across 10 ovine tissues (muscle, skin, kidney, liver, intestine, rumen, lung, testis, heart, and spleen) obtained from five individual sheep. We aimed to identify genes with stable expression across these tissues. A literature-based survey helped us shortlist candidate genes representing various functional classes from multiple livestock species. We employed four algorithms: geNorm, NormFinder, BestKeeper, and Delta Ct (ΔCt), to rank these genes based on their stability. A consistent trend in the rankings was observed across these different algorithms. RefFinder was then used for a comprehensive ranking, integrating the outputs from the various methods. ACTB, PPIB, BACH1, and B2M emerged as the most stable RGs, while RPS9, RPS15, and PGK1 displayed variable expression. We validated our findings through qPCR analysis of four target genes (ACTN2, CRYAB, DLK1, and TRIM54) in the skin samples from two different sheep breeds. Based on these results, we recommend ACTB, PPIB, BACH1, and B2M as reliable internal control genes for qPCR experiments involving diverse ovine tissues.

The present study investigated the differential expression profiles across Theileria annulata infected and non-infected crossbred cows. RNA sequencing from peripheral blood mononuclear cells (PBMCs) of four biological replicates of infected and non-infected crossbred cows generated more than 81 million reads on average across the samples. A marked difference in the expression of genes associated with innate immunity (FTH1, ACTB, ISG15) was observed between the two groups. The over-represented pathways in Theileria annulata infected cows were associated with immune system and regulation of mitotic cycle while those in non-infected animals were related to histone deacetylases (HDACs), telomerase maintenance as well as nucleosome assembly. These enriched pathways suggest maintenance of chromatin integrity which may play a protective role in the host cell. The top ten highly connected genes in infected animals were involved in interferon signaling and cell cycle, while those in non-infected animals were associated with histones or cell structure. The highly connected genes identified in this study form potential candidates for further investigation into host-parasite interactions in cattle. An improved understanding of the host response to theileriosis would lead to better management and control of the infection.

Despite immense contribution of buffaloes as dairy species, limited studies have addressed the bubaline mastitis as compared to cattle. This was the first differential transcriptomic study investigating the alterations induced by clinical mastitis in buffalo milk relative to healthy controls. Comparative gene expression profiling of three biological replicates of each group identified 1014 up-regulated and 999 down-regulated genes in the diseased buffaloes (Fold change > 2, FDR < 0.05). Activation of immune and inflammatory responses were the most enriched GO terms in the mastitic animals, with higher transcript abundance of many genes coding for anti-microbial proteins such as β-defensins, perforin, granzymes, granulysin, cathelicidins etc. Analysis of the gene regulatory interactions of the up-regulated DEGs identified many hub genes that govern the cellular and macromolecular metabolic processes (E2F4, E2F1, RBL2, FOXM1, IRF1 and MYB). This study contributes to an insightful understanding of molecular mechanisms governing immune response of buffaloes to mastitis.

Theileriosis is one of the top ten economically important diseases in cattle in India. Cytokines are considered important mediators and regulators of the immune response to an infection. In the present study, the gene expression profiles of fourteen cytokines (IL1A, IL1B, IL2, IL4, IL6, IL8, IL10, IL12A, IL12B, IL16, TGFB1, TNFA, IFNA and IFNB) were compared in Theileria annulata–infected and healthy crossbred cattle. Blood samples were obtained from the District Disease Diagnostic Laboratory, Karnal. The presence/absence of T. annulata infection in the animals was determined on the basis of blood smear examination and molecular detection through PCR using the genus-specific primers. Total RNA was extracted from peripheral blood mononuclear cells, which was further reverse transcribed to cDNA. Primer3 software was employed to design the primers for Real-Time qPCR. The results were examined using 2–∆∆Ct method with RPS15 and GAPDH as the reference genes. The expression of IL1B, IL6, IL8, IL10, IL12A, IL12B, TNFA, IFNA and IFNB was significantly higher, whereas the expression of IL2 was lower in the infected animals. The transcript levels of IL1A and TGFB1 were also higher in the diseased animals, but the results were non-significant. This study profiles the expression kinetics of various pro-inflammatory and anti-inflammatory cytokine genes in response to bovine theileriosis.

Background: Stress at workplace keep individual at extreme pressure and leads to exhaustion and inability to deal with stressful situation which causes burnout. Continuous stress on workplace causes multiple physical as well as psychological manifestations. Due to more demanding job nurses are more prone to burnout syndrome at workplace. Early assessment and timely interventions to overcome burnout are key for more productive work output. The objective of the present study was to develop Burnout Syndrome Assessment Scale (BOSAS) for nurses. Materials and Methods: A methodological design was carried out to develop BOSAS in different phases. Modified Delphi technique used for content validity. Reliability was calculated in terms of internal stability and stability of scale. Validity was measured in terms of face, content, criterion, and construct validity. Results: There were a total of 20 items in the final draft. Internal consistency was checked by Cronbach's alpha which was 0.94. Test-retest reliability of scale was found 0.93. Pearson's correlation was used to check inter-item correlation which was found between 0.20 and 0.40. The content validity index was 0.93. Concurrent criterion validity of scale was 0.82. Construct validity was analyzed by factor analysis, all items were loaded in 5 factors and accounted for 64.95% of variance. Conclusion: BOSAS has good validity and reliability and it can be used to assess and identify burnout among nurses.

In this study, changes in expression profiles of genes encoding 14 cytokines (IL1A, IL1B, IL2, IL4, IL6, IL8, IL10, IL12A, IL12B, IL16, IFNA, IFNB, TGFB1, and TNFA) were investigated amongst six Anaplasma marginale infected and six healthy crossbred cattle. Health status of the animals was determined based on clinical signs, blood smear examination and molecular detection using A. marginale-specific primers. Total RNA was isolated from the peripheral blood mononuclear cells of the infected animals as well as the healthy controls, which was further reverse transcribed to cDNA. Primers for real time PCR were designed using Primer3 software and the results were analyzed by the 2-ΔΔCt method with RPS15 and GAPDH as the reference genes. The expression levels of IL1A, IL1B, IL6, IL10, IL12A, IL12B, and TNFA varied significantly between the two groups, with higher expression in the infected cattle. The transcript abundance of IL4, IL16, and TGFB1 did not vary between the diseased and healthy animals. The expression of IL2 and IL8 was higher in the healthy animals, but the results were non-significant. Taken together, this study provides evidence for difference in expression of cytokine genes in response to anaplasmosis in crossbred cattle.

BACKGROUND: The nationwide coronavirus (COVID-19) pandemic and ensuing lockdown has enforced institutions crosswise India to provisionally close to inhibit the spread of the virus and started online learning for students. To measure the level of satisfaction of nursing students with online learning and to identify the barriers which restrict to online learning. MATERIALS AND METHODS: The current study adopted quantitative research approach with an online survey research design and carried out during May–June 2020. Participants were selected through a web-based survey (Google form), in which 219 students enrolled. Self-structured questionnaire with the Likert scale was used to measure the level of satisfaction of nursing students with online learning and identify the barriers which restrict online learning. The descriptive and inferential statistics were used for the analysis in which 219 participants were enrolled in the study of data with IBM SPSS version 20. RESULTS: Majority of student's participants 148 (67.57%) were extremely satisfied with online learning. The findings suggest that the highest barriers which restrict to online learning among nursing students is low voice and language clarity (2.16 ± 0.593), physical health barriers such as eye strain (2.43 ± 0.613), reliability and connectivity problem (2.26 ± 0.534). Among all demographic data, age is significantly associated with the level of satisfaction of online learning. CONCLUSIONS: The study data indicated that maximum students were extremely satisfied the with online learning and among barriers which effect online learning is low voice and language clarity, reliability and connectivity problem, physical health barriers such as eye strain.

Background: Coronavirus spreads very rapidly and nursing staff is fighting like warrior. They are facing so many psychological issues. During the pandemic, positive and negative emotions of frontline nurses interlace and exist. Objectives: To explore the psychological experience of nursing officers caring coronavirus disease 2019 (COVID-19) patients. Materials and Methods: Phenomenological research approach was adopted. Twelve nursing officers were enrolled with nonprobability purposive sampling technique who provided care for COVID-19 patients. The interviews were conducted face-to-face and were analyzed by the Colaizzi's 7-step method. Results: The data were obtained from 12 nursing officers caring for patients with COVID-19. The mean age of the participants was 28 ± 1.87 years, with a mean work experience of 3.75 ± 1.05. Three themes were extracted from the detailed interview. First theme: the expression of negative emotions in the initial stage consisting of fear and anxiety regarding infection and family member, fatigue and discomfort related to high work intensity, and protective gear and anxiety regarding changing of ward and long duty hours. Second theme: the use of coping strategies and self-care styles consisting of communication with family members and friends and adjustment to the new era of life. Third theme: expressing positive emotions as adjusted with duty comprised of motivation and support from senior authority, feeling happy to be a part of humanity service, and satisfied with my duty and postduty quarantine facilities. Conclusions: The results of this study show that nursing officers caring COVID-19 patients are experiencing mental and emotional distress at initial stage of duty. As days passed participants used coping strategies and understand the new era of situation. Gradually, they expressed positive emotions and they were happy and satisfied with work.

Stock prices change everyday by market forces (supply and demand). In recent years stock price prediction has been one of the most significant concern. Investors are investing on stock market on the basis of certain prediction. For prediction, stock market prices investors are applying some techniques and methods through which they get more profits and minimize their risks. Machine Learning methods are often used for the prediction of stock prices. This survey paper discusses various machine learning approaches (Supervised or Unsupervised) and methods through which the investors get to know the stock prices increase or decrease. It was done in five phases, such as data acquired, pre-processing of dataset, extraction of features, prediction of stock price using different techniques and display the result. In first phase, the data is collected from different social sites, historical data of companies. In second phase, the removal of incorrect, duplicate and dirt is done in pre-processing phase. In third phase, the reduction of data sets and the selection of useful data is done. In fourth phase, prediction is done using different machine learning techniques and approaches which is categorized as supervised and unsupervised learning techniques. Now, in last phase the accuracy is determined using different approaches.

Online platform working, Gig working, E lancing and Moonlighting have become synonymous in the Industry 4.0. Searching for and practicing alternative employments is an important phase in recording the sequence of employees’ withdrawal cognitions (WC). WC have been studied in the past majorly in relation with job attitudes for the ultimate consequence of turnover but in this virtual era, it is equally considerable to investigate the relationship of job attitudes with this important cognition sequenced before turnover stage, i.e. moonlighting, which may ultimately lead to turnover. This paper proposes to conduit the research gap of investigating the effect of Job Satisfaction on Moonlighting Intentions and the mediating effect of Organizational Commitment between these two variables. SmartPLS 3.0 software has been used to execute partial least squares structural equation modeling (PLS-SEM) for the research framework on a sample of 161 IT professionals working in the North Indian IT hubs of N.C.R. Delhi and Chandigarh. In the present study, four hypotheses have been employed for examining the relationships among the variables Job Satisfaction, Organizational Commitment and Moonlighting Intentions. PLS –SEM supported all of them. Organizational Commitment has a mediating effect between Job Satisfaction & Moonlighting Intentions while Job Satisfaction puts a very high positive impact on Organizational Commitment. In addition, Organizational Commitment shows a very significant inversely proportionate relationship with Moonlighting Intentions.

The stock market prices change everyday by market forces (supply and demand). In recent years stock market forecasting becomes a successful approach to predict stock prices. Investors are investing in the stock market based on certain predictions. For guiding stock market investors, this research paper proposes a NLP and LSTM based forecasting model for stock market. According to our research, we found a strong bond between social media news and historical data. An algorithm is proposed for sentimental analysis to establish the correlation between the stock market values and the sentiments in news feed. In our model we have utilized two different methods Natural language processing (NLP) for the feature extraction and Long Short-Term Memory (LSTM) for training our dataset. The simulated experiment based performance evaluation and comparison of the proposed model outperformed the state-of-the-arts by achieving high prediction accuracy by reducing the mean square error up to 0.062.

Workaround in the hospital setting acts as a dichotomous trope. These behaviours entitle, yet compromise the quality care to the patients. These behaviours provide and hide information about medical team members’ work. Workarounds can be individual or collective. As nurses make the majority of the health care workforce, it is essential to understand the use of workarounds in this group. Analysing and understanding the nurses’ practice and their perception of workaround behaviours is central to improving quality patient care at the bedside, where care is delivered to the patients. To exhaust the negative and tackle the positive potential of workarounds in hospitals, the management must acknowledge the factors that in uence implementation and proliferation of workarounds and the role of local and organisational culture in resolving these behaviours.

Every human being wants to live and enjoy the fruits of life till he dies. But sometimes a human being wishes to earnestly end his life by use of unnatural means. This will be abnormal to end one’s life in such way. Nursing staff spend most of time with patient at end of life so they can understand how much painful it is. The present study was conducted to assess the attitude towards euthanasia among nursing ofÀcers working in ICUs. In this study, cross sectional descriptive research survey design was adopted. Participants were selected through a non-probability convenient sampling technique, in which 90 Nursing ofÀcers working in ICUs were enrolled. Euthanasia Attitude Scale which has 4 domains and each domain is having some items (total 20 items) with the 5-point Likert scale (strongly disagree, disagree, neutral, agree, strongly Agree) to assess the attitude of Nursing ofÀcers. The study revealed that out of 90 nursing ofÀ cers 47 (52.22%) had Neutral attitude and 43 (47.77%) had Favourable attitude towards euthanasia. No association was found between participants’ attitude and demographic variables. Most of the nursing ofÀcers had neutral attitude toward euthanasia. We recommend that same study may be conducted in different settings to know the attitude of nursing ofÀcers who are working in general ward and unit.