OBJECTIVE: To investigated the dynamic ventilatory responses and their influence on functional exercise capacity in patients with long-COVID-19 syndrome (LCS). RESULTS: Sixteen LCS patients were subjected to resting lung function (spirometry and respiratory oscillometry-RO) and cardiopulmonary performance to exercise (Spiropalm®-equipped six-minute walk test-6MWT and cardiopulmonary exercise test-CPX). At rest, spirometry showed a normal, restrictive and obstructive pattern in 87.5%, 6.25% and 6.25% of participants, respectively. At rest, RO showed increased resonance frequency, increased integrated low-frequency reactance and increased difference between resistance at 4-20 Hz (R4-R20) in 43.7%, 50%, and 31.2% of participants, respectively. The median of six-minute walking distance (DTC6) was 434 (386-478) m, which corresponds to a value of 83% (78-97%) of predicted. Dynamic hyperinflation (DH) and reduced breathing reserve (BR) were detected in 62.5% and 12.5% of participants, respectively. At CPX, the median peak oxygen uptake (VO2peak) was 19 (14-37) ml/kg/min. There was a significant correlation of 6MWD with both R4-R20 (rs=-0.499, P = 0.039) and VO2peak (rs=0.628, P = 0.009). Our results indicate that DH and low BR are contributors to poor exercise performance, which is associated with peripheral airway disease. These are promising results considering that they were achieved with simple, portable ventilatory and metabolic systems.

OBJECTIVE: Survival models are used extensively in biomedical sciences, where they allow the investigation of the effect of exposures on health outcomes. It is desirable to use diverse data sets in survival analyses, because this offers increased statistical power and generalisability of results. However, there are often challenges with bringing data together in one location or following an analysis plan and sharing results. DataSHIELD is an analysis platform that helps users to overcome these ethical, governance and process difficulties. It allows users to analyse data remotely, using functions that are built to restrict access to the detailed data items (federated analysis). Previous works have provided survival modelling functionality in DataSHIELD (dsSurvival package), but there is a requirement to provide functions that offer privacy enhancing survival curves that retain useful information. RESULTS: We introduce an enhanced version of the dsSurvival package which offers privacy enhancing survival curves for DataSHIELD. Different methods for enhancing privacy were evaluated for their effectiveness in enhancing privacy while maintaining utility. We demonstrated how our selected method could enhance privacy in different scenarios using real survival data. The details of how DataSHIELD can be used to generate survival curves can be found in the associated tutorial.

OBJECTIVE: COVID-19 mitigation measures prompted many states to revise the administration of their welfare programs. States adopted policies that varied across the U.S. to respond to the difficulties in fulfilling program requirements, as well as increased financial need. This dataset captures the changes made to Temporary Assistance for Needy Families (TANF) programs during the COVID-19 pandemic, from March 2020 through December 2020. The authors created this dataset as part of a larger study that examined the health effects of TANF policy changes during the COVID-19 pandemic. DATA DESCRIPTION: TANF is the main cash assistance program for low-income families in the U.S., but benefits are often conditional on work requirements and can be revoked if an individual is deemed noncompliant. Structural factors during the COVID-19 pandemic made meeting these criteria more difficult, so some states relaxed their rules and increased their benefits. This dataset captures 24 types of policies that state TANF programs enacted, which of the states enacted each of them, when the policies went into effect, and when applicable, when the policies ended. These data can be used to study the effects of TANF policy changes on various health and programmatic outcomes.

OBJECTIVES: In this article, a singularly perturbed delay reaction-diffusion problem with nonlocal boundary conditions is considered. The exponential fitting factor is introduced to treat the solutions inside the boundary layer which occur due to perturbation parameter. The considered problem has interior layer at [Formula: see text] and strong boundary layers at [Formula: see text] and [Formula: see text]. We proposed an exponentially fitted finite difference method to solve the considered problem. The nonlocal boundary condition is treated using Composite Simpson's [Formula: see text] rule. RESULT: The stability and uniform convergence analysis of the proposed approach are established. The error estimation of the developed method is shown to be second-order uniform convergent. Two test examples were carried out to validate the applicability of the developed numerical method. The numerical results reflect the theoretical estimations.

OBJECTIVE: COVID-19 has caused tremendous damage to U.S. public health, but COVID vaccines can effectively reduce the risk of COVID-19 infections and related mortality. Our study aimed to quantify the association between proximity to a community healthcare facility and COVID-19 related mortality after COVID vaccines became publicly available and explore how this association varied across racial and ethnic groups. RESULTS: Residents living farther from a facility had higher COVID-19-related mortality across U.S. counties. This increased mortality incidence associated with longer distances was particularly pronounced in counties with higher proportions of Black and Hispanic populations.

BACKGROUND: Next-Generation Sequencing (NGS)-based testing in cancer patients has led to increased detection of variants of uncertain significance (VUS). VUS are genetic variants whose impact on protein function is unknown. VUS pose a challenge to clinicians and patients due to uncertainty regarding their cancer predisposition risk. Paucity of data exists on the pattern of VUS in under-represented populations. This study describes the frequency of germline VUS and clinico-pathological features in Sri Lankan hereditary breast cancer patients. METHODS: Data of 72 hereditary breast cancer patients who underwent NGS-based testing between January 2015 and December 2021 were maintained prospectively in a database and analyzed retrospectively. Data were subjected to bioinformatics analysis and variants were classified according to international guidelines. RESULTS: Germline variants were detected in 33/72(45.8%) patients, comprising 16(48.5%) pathogenic/likely pathogenic variants and 17(51.5%) VUS. Distribution of VUS in breast cancer predisposing genes were :APC:1(5.8%), ATM:2(11.7%), BRCA1:1(5.8%), BRCA2:5(29.4%), BRIP1:1(5.8%), CDKN2A:1(5.8%), CHEK2:2(11.7%), FANC1:1(5.8%), MET:1(5.8%), STK11:1(5.8%), NF2:1(5.8%). Mean age at cancer diagnosis in patients with VUS was 51.2 years. Most common tumour histopathology was ductal carcinoma 11(78.6%). 50% of tumours in patients having VUS in BRCA1/2 genes were hormone receptor negative. 73.3% patients had family history of breast cancer. CONCLUSIONS: A significant portion of patients had a germline VUS. Highest frequency was in BRCA2 gene. Majority had family history of breast cancer. This highlights the need to undertake functional genomic studies to determine the biological effects of VUS and identify potentially clinically actionable variants that would be useful for decision-making and patient management.

OBJECTIVES: Mechanical loading is crucial for tendon matrix homeostasis. Under-stimulation of tendon tissue promotes matrix degradation and ultimately tendon failure. In this study, we examined the expression of tendon matrix molecules and matrix-degrading enzymes (matrix metalloproteinases) in stress-deprived tail tendons and compared to tendons that were mechanically loaded by a simple restraining method. DATA DESCRIPTION: Isolated mouse tail fascicles were either floated or restrained by magnets in cell culture media for 24 h. The gene expression of tendon matrix molecules and matrix metalloproteinases in the tendon fascicles of mouse tails were examined by real-time RT-PCR. Stress deprivation of tail tendons increase Mmp3 mRNA levels. Restraining tendons represses these increases in Mmp3. The gene expression response to restraining was specific to Mmp3 at 24 h as we did not observe mRNA level changes in other matrix related genes that we examined (Col1, Col3, Tnc, Acan, and Mmp13). To elucidate, the mechanisms that may regulate load transmission in tendon tissue, we examined filamentous (F-)actin staining and nuclear morphology. As compared to stress deprived tendons, restrained tendons had greater staining for F-actin. The nuclei of restrained tendons are smaller and more elongated. These results indicate that mechanical loading regulates specific gene expression potentially through F-actin regulation of nuclear morphology. A further understanding on the mechanisms involved in regulating Mmp3 gene expression may lead to new strategies to prevent tendon degeneration.

BACKGROUND: The lack of appropriate vaccines is an obstacle to the effective management of A. baumannii infections. Peptide vaccines offer an attractive and promising preventive strategy against A. baumannii. OBJECTIVE: In this study, we identified specific T cell epitopes of A. baumannii outer membrane protein K (OMPK) using comprehensive bioinformatics and detailed molecular docking analysis. METHODS: Both class-I and class-II T cell epitopes of A. baumannii OMPK were predicted by three tools namely IEDB, SYFPEITHI, and ProPred. The predicted epitopes were shortlisted based on several analyses including prediction scoring, clustering, exclusion of human similarity, considering immunogenicity and cytokine production, and removal of toxic and/or allergen epitopes. The epitopic peptides with high prediction scores and appropriate properties containing both class-I and class-II T cell epitopes were selected. Two of these class I/II epitopic peptides were chosen for molecular docking studies and assessing their physicochemical properties as vaccine candidates. RESULTS: The results showed many T-cell epitopes of OMPK that could be evaluated for possible immunogenicity. Two of these epitopes (containing both class-I and II epitopes) had high prediction scores, were predicted by several tools, attached to several HLAs, and had the best docking score. They had different physicochemical properties and were conserved among Acinetobacter species. DISCUSSION: We identified the A. baumannii OMPK high immunogenic class-I and class-II T cell epitopes and introduced two promising high immunogenic peptides as vaccine candidates. It is recommended to perform in vitro/in vivo investigation of these peptides to determine their true efficacy and efficiency.

PURPOSE: This study investigates the applicability of optimized machine learning (ML) approach for the prediction of Medial tibial stress syndrome (MTSS) using anatomic and anthropometric predictors. METHOD: To this end, 180 recruits were enrolled in a cross-sectional study of 30 MTSS (30.36 ± 4.80 years) and 150 normal (29.70 ± 3.81 years). Twenty-five predictors/features, including demographic, anatomic, and anthropometric variables, were selected as risk factors. Bayesian optimization method was used to evaluate the most applicable machine learning algorithm with tuned hyperparameters on the training data. Three experiments were performed to handle the imbalances in the data set. The validation criteria were accuracy, sensitivity, and specificity. RESULTS: The highest performance (even 100%) was observed for the Ensemble and SVM classification models while using at least 6 and 10 most important predictors in undersampling and oversampling experiments, respectively. In the no-resampling experiment, the best performance (accuracy = 88.89%, sensitivity = 66.67%, specificity = 95.24%, and AUC = 0.8571) was achieved for the Naive Bayes classifier with the 12 most important features. CONCLUSION: The Naive Bayes, Ensemble, and SVM methods could be the primary choices to apply the machine learning approach in MTSS risk prediction. These predictive methods, alongside the eight common proposed predictors, might help to more accurately calculate the individual risk of developing MTSS at the point of care.

OBJECTIVES: To determine the effect of continuous positive airway pressure (CPAP) compliance on symptoms and signs of laryngopharyngeal reflux (LPR) in patients with obstructive sleep apnea (OSA). RESULTS: Thirty patients were included. The participants were divided into good compliance (n = 21) and poor compliance (n = 9) groups. After 6-month CPAP treatment, the reflux symptom index score significantly decreased in both the good compliance group (20 (17,24) vs. 14 (10,18), p < 0.001) and the poor compliance group (21 (18,25) vs. 10 (5,16), p < 0.05). Reflux finding score was significantly reduced in both the good compliance group (8 (6,9) vs. 4 (3,5), p < 0.001) and the poor compliance group (6 (4,8) vs. 3 (2,4), p < 0.05). However, there were no statistically significant differences between the groups. CPAP treatment reduces the symptoms and signs of LPR. However, CPAP compliance does not correlate with improvement in LPR in patients with OSA.

OBJECTIVE: The tarnished plant bug (TPB), Lygus lineolaris (Palisot de Beauvois) (Hemiptera: Miridae), is a pest damaging many cultivated crops in North America. Although partial transcriptome data are available for this pest, a genome assembly was not available for this species. This assembly of a high-quality chromosome-length genome of TPB is aimed to develop the genetic resources that can provide the foundation required for advancing research on this species. RESULTS: The initial genome of TPB assembled with paired-end nucleotide sequences generated with Illumina technology was scaffolded with Illumina HiseqX reads generated from a proximity ligated (HiC) library to obtain a high-quality genome assembly. The final assembly contained 3963 scaffolds longer than 1 kbp to yield a genome of 599.96 Mbp. The N50 of the TPB genome assembly was 35.64 Mbp and 98.68% of the genome was assembled into 17 scaffolds larger than 1 Mbp. This megabase scaffold number is the same as the number of chromosomes observed in karyotyping of this insect. The TPB genome is known to have high repetitive DNA content, and the reduced assembled genome size compared to flowcytometric estimates of approximately 860 Mbp may be due to the collapsed assembly of highly similar regions.

OBJECTIVES: The notion of Bipolarity based on positive and negative outcomes. It is well known that bipolar models give more precision, flexibility, and compatibility to the system as compared to the classical models and fuzzy models. A bipolar fuzzy graph(BFG) provides more flexibility while modeling human thinking as compared with a fuzzy graph, and an interval valued bipolar fuzzy graph(IVBFG) has numerous applications where the real-life problem are time dependent and there is a network structure complexity. The aim of this paper is to introduce an interval-valued bipolar line fuzzy graph(IVBFLG). RESULT: In this paper, we have proposed the notion of an IVBFLG and some of its characterizations. Also, some propositions and theorems related to an IVIFLGs are developed and proved. Furthermore, isomorphism between two IVIFLGs toward their IVIFGs was determined and verified. As a result, we derive a necessary and sufficient condition for an IVBFG to be isomorphic to its corresponding IVBFLG and some remarkable properties like degree, size, order, regularity, strength, and completeness of an IVBFLGs have been investigated, and the proposed concepts are illustrated with the examples.

OBJECTIVE: Active participation of the older adults in the society is crucial; however, frailty prevents social participation. Meanwhile, many older adults participate daily in social activities, even with frailty. This study aims to examine whether older adults with frailty have lower social participation than those without frailty in Japan. We also investigated whether older adults with frailty and higher subjective health participate in society to the same extent as the general older population. This study included 1,082 Japanese individuals aged 65 years and older participating in the online survey. Participants answered questions on social participation, frailty, subjective health, and demographics. RESULTS: Participants in the robust group had higher social participation rates than those in the frailty and pre-frailty groups. Meanwhile, frail older participants with higher subjective health had similar social participation as the robust participants. Many older adults acquire frailty despite their individual effort. Meanwhile, improving subjective health may be effective, even with frailty. The relationship between subjective health, frailty, and social participation is primitive and further studies are needed.

OBJECTIVE: Glomerular filtration rate (GFR) is a key indicator of renal function. In both clinical practice and pre-clinical research, serum levels of endogenous filtration markers, such as creatinine, are often used to estimate GFR. However, these markers often do not reflect minor changes in renal function. In this study, we therefore set out to evaluate the applicability of transcutaneous GFR (tGFR) measurements to monitor the changes in renal function, as compared to plasma creatinine (pCreatinine), in two models of obstructive nephropathy, namely unilateral ureteral obstruction (UUO) or bilateral ureteral obstruction followed by release (BUO-R) in male Wistar rats. RESULTS: UUO animals showed a significant reduction in tGFR compared to baseline; whereas pCreatinine levels were not significantly changed. In BUO animals, tGFR drops 24 h post BUO and remains lower upon release of the obstruction until day 11. Concomitantly, pCreatinine levels were also increased 24 h after obstruction and 24 h post release, however after 4 days, pCreatinine returned to baseline levels. In conclusion, this study revealed that the tGFR method is superior at detecting minor changes in renal function as compared to pCreatinine measurements.

OBJECTIVE: This work aimed to examine the leishmanicidal, cellular mechanisms and cytotoxicity effects of formononetin (FMN), a natural isoflavone, against Leishmania tropica. We used the MTT assay to determine the leishmanicidal effects of FMN against promastigotes and its cytotoxicity effects on J774-A1 macrophage cells. The Griess reaction assay and quantitative real-time PCR were used to determine the nitric oxide (NO) and the mRNA expression levels of IFN-γ and iNOS in infected J774-A1 macrophage cells. RESULTS: FMN significantly (P < 0.001) decreased the viability and number of promastigotes and amastigotes forms. The 50% inhibitory concentrations value for FMN and glucantime was 9.3 and 14.3 µM for promastigote and amastigote, respectively. We found that the macrophages exposed with FMN especially at concentrations of 1/2 IC50 and IC50 significantly activated the NO release and the mRNA expression levels of IFN-γ, iNOS. The findings of the current research showed the favorable antileishmanial effects formononetin, a natural isoflavone, against various stages of L. tropica through inhibition of infectivity rate of macrophage cells and triggering the NO production and cellular immunity. However, supplementary works are essential to evaluate the ability and safety of FMN in animal model before use in the clinical phase.

OBJECTIVE: The purpose of this data set is to investigate differences in RNA-Seq transcriptome profiles between Acarapis woodi-infested and uninfested Japanese honey bees (Apis cerana japonica). The data set is strengthened by data collected from different body parts (head, thorax, and abdomen). The data set will support future studies of molecular biological changes in mite-infested honey bees. DATA DESCRIPTION: We collected 5 mite-infested and 5 uninfested A. cerana japonica workers from each of 3 different colonies (designated as A, B, and C). Workers were dissected into 3 body sites (i.e., heads, thoraces, and abdomen), and 5 of each body site were pooled together for RNA extraction, generating a total of 18 RNA-Seq samples (2 infection status × 3 colonies × 3 body sites). FASTQ data files of each sample that were generated by a DNBSEQ-G400 sequencer with the 2 × 100 bp paired-end sequencing protocol are available in the DDBJ Sequence Read Archive under accession number DRA015087 (RUN: DRR415616-DRR415633, BioProject: PRJDB14726, BioSample: SAMD00554139-SAMD00554156, Experiment: DRX401183-DRX401200). The data set is a fine-scale analysis of gene expression in the mite-infested A. cerana japonica workers because 18 RNA-Seq samples are separated by 3 body sites.

OBJECTIVE: Evaluate bovine viral diarrhea virus (BVDV) antigenicity by using virus neutralization titers (VNT) analyzed using the principal component analysis (PCA) from antisera generated against US-based vaccine strains against both US-origin field isolates and non-US-origin field isolates. RESULTS: Data from both independent analyses demonstrated that several US-origin and non-US-origin BVDV field isolates appear to be antigenically divergent from the US-based vaccine strains. Results from the combined analysis provided greater insight into the antigenic diversity observed among BVDV isolates. Data from this study further support genetic assignment into BVDV subgenotypes, as well as strains within subgenotypes is not representative of antigenic relatedness. PCA highlights isolates that are antigenically divergent from members of the same species and subgenotype and conversely isolates that belong to different subgenotypes have similar antigenic characteristics when using antisera from US-based vaccine isolates.

OBJECTIVE: Cystic fibrosis (CF) is a genetic condition that causes abnormal mucus secretions in affected organs. MUC5AC and MUC5B are gel-forming mucins and frequent targets for investigations in CF tissues. Our objective was to qualify MUC5AC and MUC5B immunohistochemical techniques to provide a useful tool to identify, localize and interpret mucin expression in ferret tissues. RESULTS: MUC5AC and MUC5B mucins were detected most commonly in large airways and least in small airways, consistent with reported goblet cell density in airway surface epithelia. We evaluated whether staining method affected the detection of goblet cell mucins in serial sections of bronchial surface epithelia. Significant differences between stains were not observed suggesting common co-expression MUC5AC and MUC5B proteins in goblet cells of airway surface epithelia. Gallbladder and stomach tissues are reported to have differential mucin enrichment, so we tested these tissues in wildtype ferrets. Stomach tissues were enriched in MUC5AC and gallbladder tissues enriched in MUC5B, mucin enrichment similar to human tissues. Mucin immunostaining techniques were further qualified for specificity using lung tissue from recently generated MUC5AC-/- and MUC5B-/- ferrets. Qualified techniques for MUC5AC and MUC5B immunohistochemistry will be useful tools for mucin tissue studies in CF and other ferret models.

OBJECTIVE: Antibiotic resistant infections have become a global health crisis causing 1.2 million deaths worldwide in 2019 [1]. In a previous study, we identified a bacterium from a rare genus, Yimella, and found in an initial antibiotic screening that they produce broad-spectrum bactericidal compounds [2]. Herein, we focus on the characterization of these potential novel antimicrobial compounds produced by Yimella sp. RIT 621. RESULTS: We used solid-phase extraction and C18 reverse-phase chromatography to isolate the antibiotic-active compounds found in organic extracts from liquid cultures of Yimella sp. RIT 621. We tracked the antimicrobial activity by testing the extracts in disc diffusion inhibitory assays and observed its increase after each purification stage.

BACKGROUND: To describe the preferred cataract surgery practices among Malaysian ophthalmologists and medical officers in comparison with the recommended practices. METHODS: An online questionnaire was sent out in April 2021, to Malaysian Ophthalmologists and medical officers who perform cataract surgeries. The questions were focused on the preferred cataract surgery practices of the participants. All data obtained were collected, tabulated and analysed. RESULTS: A total of 173 participants responded to the online questionnaire. 55% of the participants were within 31-40 years of age. 56.1% preferred peristaltic pump over venturi system. 91.3% of participants practised povidone iodine instillation to the conjunctival sac. With regards to the main wound incision, more than half of the surgeons (50.3%) preferred fixed superior incision and 72.3% of them preferred 2.75 mm microkeratome blade. Most of the participants (63%) were inclined towards C-Loop clear intraocular lens (IOL) with a single-handed push preloaded system. 78.6% of the surgeons routinely use carbachol in their cataract surgery. CONCLUSIONS: This survey provides some insight into the current practices among Malaysian ophthalmologists. Most of the practices are in line with international guidelines for preventing postoperative endophthalmitis. This article could help trainees and ophthalmologists benchmark and observe the common cataract surgery practices among their seniors and peers in Malaysia.

OBJECTIVE: In Sweden, national guidelines recommend that all staff in the healthcare system systematically screen patients for alcohol use and illicit substance use. Where hazardous use is identified, it should be addressed as soon as possible, preferably through brief interventions (BI). Results from a previous national survey showed that most clinic directors stated that they had clear guidelines for screening alcohol use and illicit substance use, but that fewer staff than expected used screening in their work. This study aims to identify obstacles and solutions to screening and brief intervention, based on survey respondents' free-text responses to open-ended questions. RESULTS: A qualitative content analysis yielded four codes: guidelines, continuing education, cooperation and resources. The codes indicated that staff would need (a) clearer routines in order to optimize compliance with the national guidelines; (b) more knowledge about how to treat patients with problematic substance use; (c) better cooperation between addiction care and psychiatry; and (d) increased resources to improve routines at their own clinic. We conclude that increased resources could contribute to better routines and cooperation, and provide increased opportunities for continuing education. This could increase guideline compliance and increase healthy behavior changes among patients in psychiatry with problematic substance use.

OBJECTIVE: The microbiota of a seasoning sauce fermentation process is usually complex and includes multiple species and even various strains of one species. Moreover, composition and cell numbers of individual strains vary over the course of the entire fermentation. This study demonstrates the applicability of a multiplex PCR system to monitor growth dynamics of Tetragenococcus (T.) halophilus strains in order to evaluate their performance and help to select the most competitive starter strain. RESULTS: In a previous study we isolated T. halophilus strains from multiple lupine moromi fermentation processes and characterized them. In this study we wanted to monitor the growth dynamics of these strains in a competitive lupine moromi model fermentation process using a multiplex PCR system. Therefore, pasteurized lupine koji was inoculated with eight different T. halophilus strains, six from lupine moromi, one from an experimental buckwheat moromi fermentation process and the type strain DSM 20,339T, to create the inoculated lupine moromi pilot scale fermentation process. With the multiplex PCR system, we could detect that all strains could grow in lupine moromi but, that TMW 2.2254 and TMW 2.2264 outperformed all other strains. Both strains dominated the fermentation after three weeks with cell counts between 4 × 106 to 4 × 107 CFU/mL for TMW 2.2254 and 1 × 107 to 5 × 107 CFU/mL for TMW 2.2264. The pH dropped to value below 5 within the first 7 days, the selection of these strains might be related to their acid tolerance.

OBJECTIVE: Probiotics have been used in poultry production to improve the performance and health of chickens raised without antibiotics. The combination of different probiotic strains has been used with the hope of conferring multiple benefits to the host. However, the inclusion of several strains does not necessarily boost benefits. There is a lack of studies that compare the efficacy of multi-strain probiotics to their individual components. In this study, the effects of a Bacillus-based probiotic product mix containing B. coagulans, B. licheniformis, B. pumilus, and B. subtilis against Clostridium perfringens were tested in vitro using a co-culture method. The individual strains and different combinations of the strains used in the product were also tested against C. perfringens. RESULTS: The probiotic product mix tested in this study did not show effects against C. perfringens (P = 0.499). When tested individually, the strain of B. subtilis was the most efficient strain to decrease C. perfringens concentrations (P ≤ 0.01), and the addition of other Bacillus species strains significantly decreased its efficacy against C. perfringens. We concluded that the probiotic mix of Bacillus strains used in this study (B. coagulans, B. licheniformis, B. pumilus and B subtilis) was not effective in decreasing C. perfringens concentrations in vitro. However, when deconstructing the probiotic, the strain of B. subtilis alone or combined with the strain of B. licheniformis were effective against C. perfringens. This suggests that the anticlostridial properties of the particular strains of Bacillus used in this study were negatively affected when combined with other Bacillus spp. strains.

OBJECTIVE: The paper is focused on developing and analyzing a uniformly convergent numerical scheme for a singularly perturbed reaction-diffusion problem with a negative shift. The solution of such problem exhibits strong boundary layers at the two ends of the domain due to the influence of the perturbation parameter, and the term with negative shift causes interior layer. The rapidly changing behavior of the solution in the layers brings significant difficulties in solving the problem analytically. We have treated the problem by proposing a numerical scheme using the implicit Euler method in the temporal direction and a fitted tension spline method in the spatial direction with uniform meshes. RESULT: Stability and uniform error estimates are investigated for the developed numerical scheme. The theoretical finding is demonstrated by numerical examples. It is obtained that the developed numerical scheme is uniformly convergent of order one in time and order two in space.

OBJECTIVE: Chalcone synthase (CHS) catalyzes the initial step of the flavonoid biosynthesis. The CHS encoding gene is well studied in numerous plant species. Rapidly growing sequence databases contain hundreds of CHS entries that are the result of automatic annotation. In this study, we evaluated apparent multiplication of CHS domains in CHS gene models of four plant species. MAIN FINDINGS: CHS genes with an apparent triplication of the CHS domain encoding part were discovered through database searches. Such genes were found in Macadamia integrifolia, Musa balbisiana, Musa troglodytarum, and Nymphaea colorata. A manual inspection of the CHS gene models in these four species with massive RNA-seq data suggests that these gene models are the result of artificial fusions in the annotation process. While there are hundreds of seemingly correct CHS records in the databases, it is not clear why these annotation artifacts appeared.

OBJECTIVE: The green fluorescent protein (GFP) and its derivatives are widely used in biomedical research. The manipulation of GFP-tagged proteins by GFP-specific binders, e.g. single-domain antibodies (nanobodies), is of increasing significance. It is therefore important to better understand the properties of antiGFP-GFP interaction in order to establish methodological applications. In this work the interaction of superfolder GFP (sfGFP) and its enhancer nanobody (aGFPenh) was characterized further. RESULTS: Previous calorimetric experiments demonstrated that the aGFPenh nanobody binds strongly to sfGFP with a nanomolar affinity. Here we show that this interaction results in a substantial structural stabilization of aGFPenh reflected in a significant increase of its melting temperature by almost 30 °C. The thermal stability of the sfGFP-aGFPenh complex is close to 85 °C in the pH range 7.0-8.5. For therapeutic applications thermoresistance is often an essential factor. Our results suggest that methodologies based on GFP-aGFP interaction can be applied under a wide range of physicochemical conditions. The aGFPenh nanobody seems to be suitable for manipulating sfGFP-labeled targets even in extreme thermophilic organisms.

OBJECTIVE: A mammalian Delta-Notch signaling component, Notch1, has been suggested for its expression during the normal sperm development although its conditional deletion caused no apparent abnormalities. Since we established our original transgenic mouse system that enabled labeling of past and ongoing Notch1 signaling at a cellular level, we tried to validate that observation in vivo. Our transgenic mouse system used Cre/loxP system to induce tandem dsRed expression upon Notch1 signaling. RESULTS: To our surprise, we were unable to observe tandem dsRed expression in the seminiferous tubules where the sperms developed. In addition, tandem dsRed expression was lacking in the somatic cells of the next generation in our transgenic mouse system, suggesting that sperms received no Notch1 signaling during their development. To validate this result, we conducted re-analysis of four single-cell RNA-seq datasets from mouse and human testes and showed that Notch1 expression was little in the sperm cell lineage. Collectively, our results posed a question into the involvement of Notch1 in the normal sperm development although this observation may help the interpretation of the previous result that Notch1 conditional deletion caused no apparent abnormalities in murine spermatogenesis.

OBJECTIVE: Phenotypes and mechanisms of cardiovascular disease (CVD) may differ across global populations. In sub-Saharan Africa (SSA), distinct environmental determinants may influence development and progression of atherosclerotic coronary artery disease (CAD). METHODS: We investigated associations between 6 established markers of myocardial stress and subsequent subclinical CAD (sCAD), defined as presence of any atherosclerosis on coronary CT angiography (CCTA) in a 2-year prospective cohort of Ugandan adults enriched for cardiometabolic risk factors (RFs) and HIV. Six plasma biomarkers were measured baseline among 200 participants (50% with HIV) aged ≥ 45 years with ≥ 1 cardiovascular RF. At 2-year follow-up, 132 participants (52% with HIV) who returned underwent coronary CCTA. RESULTS: In logistic regression models adjusted for cardiovascular RFs (age, diabetes, hypertension, hyperlipidemia, smoking, obesity) and non-traditional RFs (HIV, chronic kidney disease), only NT-proBNP predicted subsequent subclinical CAD (p < 0.008, Bonferroni correction for multiple testing). In sensitivity analyses adjusted for ASCVD risk category (instead of individual RFs) in the baseline cohort with multiple imputation applied to missing year 2 CCTA data (n = 200), NT-proBNP remained significantly associated with subsequent CAD (p < 0.008). CONCLUSIONS: NT-proBNP consistently predicted subclinical CAD in Uganda in the absence of such an association among other markers of myocardial stress, suggesting a role for NT-proBNP in atherosclerosis independently of coronary microvascular dysfunction.

AIM: This cross-sectional survey aimed to identify aerobic bacteria, antimicrobial resistance, and multi-drug resistance profiles of bacteria isolated from different wound infections among a group of Egyptian patients. RESULTS: Of 120 positive samples, 170 isolates were identified. Polymicrobial infections were determined in 55% of samples. The dominant Gram-positive isolated strains were Staphylococcus aureus, especially from wound infections because of accidents (71.8%). Piperacillin, methicillin, ampicillin/sulbactam, and amoxicillin/clavulanic acid were all highly resistant to S. aureus and Coagulase-negative Staphylococci. The prevalence of methicillin-resistant S. aureus in wound infections was 89.9%. S. aureus showed superior sensitivity to vancomycin (85.3%) and linezolid (81.3%). The highest prevalence of Gram-negative isolates was for Pseudomonas aeruginosa (40%), which was highly sensitive to ciprofloxacin (79.2%) and highly resistant to levofloxacin (83.3%). Several isolates revealed a multi-drug resistance profile (52.4%). The overall MDR rate of Gram-positive and Gram-negative isolates were 50% and 54.9%, respectively. CONCLUSION: The prevalence of MRSA isolated from various wound infections and MDR is a warning issue in Upper Egypt. It should implement a health education strategy and hygiene measures to prevent the spread of wound infection-causing organisms in the community.

OBJECTIVE: To investigate CITED1 as a potential biomarker of anti-endocrine response and breast cancer recurrence, given its previously determined role in mediating estrogen-dependant transcription. The study is a continuation of earlier work establishing the role of CITED1 in mammary gland development. RESULTS: CITED1 mRNA is associated with estrogen-receptor positivity and selectively expressed in the GOBO dataset of cell lines and tumours representing the luminal-molecular subtype. In patients treated with tamoxifen, higher CITED1 correlated with better outcome, suggesting a role in anti-estrogen response. The effect was particularly evident in the subset of estrogen-receptor positive, lymph-node negative (ER+/LN-) patients although noticeable divergence of the groups was apparent only after five years. Tissue microarray (TMA) analysis further validated the association of CITED1 protein, by immunohistochemistry, with favourable outcome in ER+, tamoxifen-treated patients. Although we also found a favourable response to anti-endocrine treatment in a larger TCGA dataset, the tamoxifen-specific effect was not replicated. Finally, MCF7 cells overexpressing CITED1 showed selective amplification of AREG but not TGFα suggesting that maintenance of specific ERα-CITED1 mediated transcription is important for the long-term response to anti-endocrine therapy. These findings together confirm the proposed mechanism of action of CITED1 and support its potential use as a prognostic biomarker.

OBJECTIVES: The purposes of this study were to determine the Efficiency of Plating (EOP) value of Bacteriophage BI-EHEC and BI-EPEC and to evaluate the application of these bacteriophages in reducing population of EHEC and EPEC on various food samples. RESULTS: In this study, we used bacteriophage BI-EHEC and BI-EPEC, which were isolated from previous study. Both phages were tested with other multiple pathotypes of intestinal pathogenic E. coli to determine the efficiency of plating. BI-EHEC had high efficiency toward ETEC with an EOP value of 2.95 but low efficiency toward EHEC with an EOP value of 0.10, while BI-EPEC had high efficiency toward EHEC and ETEC with EOP values of 1.10 and 1.21, respectively. As biocontrol agents, both bacteriophages able to reduce CFU of EHEC and EPEC in several food samples using 1 and 6-days incubation times at 4 [Formula: see text]. BI-EHEC reduced the number of EHEC with an overall percentage of bacterial reduction value above 0.13 log10, while BI-EPEC reduced number of EPEC with reduction value above 0.33 log10.

OBJECTIVE: Bananas are one of the most popular fruits in the world, providing food security and employment opportunities in several developing countries. Increasing the anthocyanin content of banana fruit could improve the health-promoting properties. Anthocyanin biosynthesis is largely regulated at the transcriptional level. However, relatively little is known about the transcriptional activation of anthocyanin biosynthesis in banana. RESULTS: We analysed the regulatory activity of three Musa acuminata MYBs that were predicted by bioinformatic analysis to transcriptionally regulate anthocyanin biosynthesis in banana. MaMYBA1, MaMYBA2 and MaMYBPA2 did not complement the anthocyanin-deficient phenotype of the Arabidopsis thaliana pap1/pap2 mutant. However, co-transfection experiments in A. thaliana protoplasts showed that MaMYBA1, MaMYBA2 and MaMYBPA2 function as components of a transcription factor complex with a bHLH and WD40 protein, the so called MBW complex, resulting in the activation of the A. thaliana ANTHOCYANIDIN SYNTHASE and DIHYDROFLAVONOL 4-REDUCTASE promoters. The activation potential of MaMYBA1, MaMYBA2 and MaMYBPA2 was increased when combined with the monocot Zea mays bHLH ZmR instead of the dicot AtEGL3. This work paves the path towards decoding the MBW complex-mediated transcriptional activation of anthocyanin biosynthesis in banana. It will also facilitate research towards increased anthocyanin content in banana and other monocot crops.

OBJECTIVES: Warfarin, like many other anticoagulants, has been linked to an elevated risk of bleeding proportional to the amount of anticoagulation used. Not only was the incidence of bleeding raised by the dosage, but the subtherapeutic international normalized ratio (INR) was also associated with increased thrombotic events. This retrospective cohort and multi-center study evaluated the incidence and risk factors of warfarin therapy complications in community hospitals in Thailand's central and eastern regions from 2016 to 2021. RESULTS: Among 335 patients (683.90 person-year of follow-up), The incidence rate of warfarin complications was 4.91 events per 100 person-year. The independent factor associated with warfarin therapy complications was propranolol prescription (Adjusted RR: 2.29, 95%CI: 1.12-4.71). The secondary analysis was divided according to the outcome of the major bleeding and thromboembolic event. Major bleeding events, hypertension (Adjusted RR: 0.40, 95%CI: 0.17-0.95), amiodarone prescription (Adjusted RR: 5.11, 95%CI: 1.08-24.15), and propranolol prescription (Adjusted RR: 2.86, 95%CI: 1.19-6.83) were the independent risk factors. While in the major thrombotic event, non-steroidal anti-inflammatory drugs (NSAIDs) prescription was an independent factor (Adjusted RR: 10.65, 95%CI: 1.26-90.35).

OBJECTIVE: Prevention of high-risk behaviors has always been considered; According to the researches, a person's religious attitudes and beliefs and intelligence quotient are associated with the prevention of high-risk behaviors such as drug addiction, and religiosity and spirituality also reduce drug addiction; Therefore, this study was conducted to compare religious beliefs, intelligence and spiritual health in two methods of education-based treatment and methadone in people with addiction. RESULTS: A comparative study was performed among 184 people on all drug users admitted to these wards that treated with methadone and participants of meetings of anonymous drug users. Four questionnaires were used to collect information. Mean, and standard deviation were used to describe the demographic characteristics of participants. Chi-square and fisher tests were used to compare demographic information in the two groups. The present study was performed following acquisition of the code of ethics (IR.BUMS.REC.1395.156) from Research Ethics Committee of Birjand University of Medical Sciences.

OBJECTIVE: We present 16s rRNA gene sequencing (V1-V2 region) and sample data from a pilot observational cohort study to describe the gut microbiota dynamics of subjects with latent tuberculosis infection (LTBI) treated with daily 600 mg rifampicin for four months (4R) or a weekly dose of 900 mg combination of rifapentine and isoniazid for three months (3HP). Our objectives were to (1) document changes to the gut microbiota immediately following exposure to the rifamycins and (2) document recovery to baseline two months after treatment completion. DATA DESCRIPTION: We enrolled six subjects with subjects with LTBI and prospectively followed them for 5-6 months. Each subject provided stool samples before, during, and two months after treatment. Six healthy controls were sampled in parallel with the patients with LTBIs. We report amplicon sequence variants (ASVs) and taxonomic assignments for 60 stool samples. Additionally, we provide access to the raw amplicon sequences, and subject responses to questionnaires about their diet, medication, and lifestyle changes over the study follow-up period. Furthermore, we provide the concentration of the parent and partially active rifamycin metabolite concentrations measured validated LC-MS-MS assays of phosphate buffer washes of the stool samples collected from the LTBI participants. This comprehensive dataset is a valuable resource for future systematic reviews and meta-analyses of the impact of LTBI therapy on the gut microbiota.

OBJECTIVE: Retinoic acid (RA) is known to transition proliferating SH-SY5Y neuroblastoma cells towards functional neurons. However, the activity of RA is restricted due to its photolability where any findings from prolonged time course observations using microscopy may alter outcomes. The aim of the study was to establish a real-time, long-term (9-day) protocol for the screening of differentiation events using Electrical cell-substrate impedance sensing (ECIS). RESULTS AND DISCUSSION: A differentiation baseline for SH-SY5Y cells was established. Cells were seeded and exposed to repeated spikes of RA using the xCELLigence real-time cell analyser single plate (RTCA-SP) for real-time monitoring and identification of differentiation activity over a 9 day period in order to be more representative of differentiation over a prolonged timeline. Specific features associated with differentiation (growth inhibition, neurite outgrowths) were confirmed by end-point analysis. RA-induced growth inhibition and assumed phenotypic changes (i.e. neurite outgrowth) were identified by the xCELLigence analysis and further confirmed by end-point metabolic and phenotypic assays. Change in cellular morphology and neurite outgrowth length was identified by end-point fluorescence detection followed by computational analysis. Based on this it was possible to identify SH-SY5Y phenotypic differentiation with distinct phases observed over 9 days using Electric cell-substrate impedance sensing (ECIS) cell index traces providing a path to application in larger scale neurotrophic factor screening using this scalable technology.