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Predictive designs to recognize Holstein cattle vulnerable to metritis and also medical heal and reproductive/productive failing following anti-microbial treatment.
better understand the potential risks and outcome in patients exhibiting early radiographic changes following chemoradiation.Bacillus velezensis FTL7 which exhibited potent antimicrobial peptide producing capacity was isolated from a marine sediment sample of the West Coast region, South India, and characterized through experimental and genomic analysis approaches. FTL7 showed potential antimicrobial activity against a broad range of foodborne pathogenic bacteria like Listeria monocytogenes Scott A, Bacillus cereus (ATCC 11778), Salmonella Typhimurium (MTCC 1251), Staphylococcus aureus (ATCC 25923), and Escherichia coli (MTCC 443). It also exhibited strong inhibitory activity against Kocuria rhyzophila (ATCC 934) and Bacillus subtilis subsp. spizizenii (ATCC 6633). Phylogenetic analysis by 16S rRNA gene sequence showed that Bacillus velezensis FTL7 was closely related to B. velezensis LBUM288 (GenBank accession number MG461457) with 100% identity. Whole-genome sequencing of the strain FTL7 was carried out using Illumina sequencing technology to get a better insight into the mechanisms of controlling pathogens by FTL7. The strain FTL7 has a chromosome size of 3849,077 bp with a GC content of 46.56%. The genome consists of 3635 coding sequences, 64 RNA, 59 tRNAs, 5 ncRNAs, and 69 pseudogenes. The presence of genes responsible for the synthesis of non-ribosomal peptides and bacteriocins was identified through genome annotation. Thus, many Bacillus strains, including B. velezensis, have been demonstrated as excellent producers of antimicrobial substances.
Guidance for dabigatran and rivaroxaban in overweight patients diagnosed with non-valvular atrial fibrillation (NVAF) is still lacking.

Compare the effectiveness and safety of dabigatran and rivaroxaban for the treatment of NVAF in the overweight population.

A total of 396 out of 1029 overweight patients with NVAF at Zhongshan Hospital, Fudan University, from January 2017 and December 2018 were retrospectively enrolled using propensity score matching analysis. The clinical outcomes were analyzed by chi-square test and Kaplan-Meier analyses. The risk of bleeding and thrombosis was assessed using a Cox regression analysis and validated using a nomogram model.

In terms of effectiveness, the incidence of thrombosis events and the time to thrombosis were similar in the dabigatran and rivaroxaban groups (P > 0.05). Regarding safety, compared to dabigatran, the rivaroxaban group had a higher incidence of bleeding events (8.6% vs. 3.5%, χ
 = 4.435, P = 0.035), a shorter time to bleeding (11.3 ± 0.18months vs. 11.6 ± 0.14months, P = 0.038) and an increased risk of bleeding (hazard ratio HR = 2.452, 95% confidence interval CI 1.017-5.913, P = 0.046), especially in those patients with heart failure (HR = 3.207, 95% CI 1.183-8.694, P = 0.022).

Dabigatran therapy was shown to be equally effective. It may be superior in reducing bleeding risk in an overweight population with NVAF than rivaroxaban. Further prospective studies are encouraged for analysis.
Dabigatran therapy was shown to be equally effective. It may be superior in reducing bleeding risk in an overweight population with NVAF than rivaroxaban. Further prospective studies are encouraged for analysis.Motile cilia are hair-like structures that move and propel fluid, playing important roles in the physiology of organs. Here, we present a protocol to visualize and measure ciliary beating and cerebrospinal fluid (CSF) flow in the telencephalon of an adult zebrafish brain explant. We describe the preparation of brain explants, the recording of ciliary beating and CSF flow, and data analysis using ImageJ and MATLAB. These imaging and analysis techniques can be directly translated to other ciliated systems. For complete details on the use and execution of this protocol, please refer to D'Gama et al. (2021).Phosphoinositides (PIPs) are low-abundant membrane lipids with critically important functions in cellular physiology. To investigate their subcellular distribution in neurons, we optimized protocols for immunofluorescence staining of intracellular or plasma membrane PIPs with commercially available antibodies. Here, we describe the preparation and transfection of primary mouse hippocampal neurons in dissociated culture, followed by immunofluorescence staining and quantitative analysis of PIP signals. In addition, we expand the application of the protocol to proteins located at the cytoplasmic leaflet of cellular membranes. For complete details on the use and execution of this protocol, please refer to Guo et al. (2022).Our recent development of the CRAGE (chassis-independent recombinase-assisted genome engineering) system enables single-step integration of large, complex DNA constructs directly into bacteria genomes across multiple phyla. Guanosine datasheet This protocol describes the details of the experimental design and procedures of CRAGE and extended CRAGE-Duet systems. It also describes a strategy that combines CRISPR with CRAGE, which allows implementation of CRISPR-Cas9, CRISPRa, and CRISPRi in diverse bacteria, overcoming major limitations to broaden the application of CRISPR in non-model bacterial genome engineering. For complete details on the use and execution of this protocol, please refer to Wang et al. (2019), Wang et al. (2020), and Liu et al. (2020).The suprachiasmatic nucleus (SCN) is the master circadian pacemaker of the mammalian biological clock. Here, we provide a detailed protocol for long-term recording of calcium signals in SCN neurons of freely moving mice through a multichannel optical fiber recording system. This system can simultaneously collect calcium signals from up to seven animals. The calcium signals can be visualized by the appropriate software and code. This protocol can be used to explore the long-term response of SCN to external environmental stimulation. For complete details on the use and execution of this protocol, please refer to Zhai et al. (2022).Clinically relevant animal models are crucial for effective development of therapeutics for peritoneal carcinomatosis (PC). This protocol describes the generation of patient-derived ascites-dependent xenograft (PDADX) models from the cellular component of ascites. The use of routine intraperitoneal injection of the fluid component of ascites is analogous to the biological events occurring intra-abdominally in patients with PC. By serving as a proxy, PDADX models represent a valuable tool for preclinical testing of new therapeutics for PC. For complete details on the use and execution of this protocol, please refer to Hendrikson et al. (2022).Male germ-cell development comprises primordial germ-cell (PGC) development, spermatogonium differentiation, and ensuing spermatogenesis. We present a step-by-step protocol for differentiation of mouse pluripotent stem cells (PSCs) into germline stem-cell-like cells (GSCLCs) via PGC-like cell and spermatogonium-like cell intermediates. The differentiation protocol has higher fidelity than our previous protocol. Upon transplantation into testes in vivo or culture for testis transplants, GSCLCs robustly contribute to spermatogenesis, providing a paradigm for PSC-based reconstitution of mammalian male germ-cell development. For complete details on the use and execution of this protocol, please refer to Ishikura et al. (2021).
Data-dependent acquisition (DDA) is the most commonly used MS/MS scan method for lipidomics analysis on orbitrap-based instrument. However, MS instrument associated software decide the top N precursors for fragmentation, resulting in stochasticity of precursor selection and compromised consistency and reproducibility. We introduce a novel workflow using biologically relevant lipids to construct inclusion list for data-independent acquisition (DIA), named as BRI-DIA workflow.

To ensure consistent coverage of biologically relevant lipids in LC-MS/MS-based lipidomics analysis.

Biologically relevant ion list was constructed based on LIPID MAPS and lipidome atlas in MS-DIAL 4. Lipids were extracted from mouse tissues and used to assess different MS/MS scan workflow (DDA, BRI-DIA, and hybrid mode) on LC-Orbitrap Exploris 480 mass spectrometer.

DDA resulted in more MS/MS events, but the total number of unique lipids identified by three methods (DDA, BRI-DIA, and hybrid MS/MS scan mode) is comparable (580 uninal DDA method in profiling lipids, but offers better consistency of lipid identification, compared to DDA method. This study was performed using Orbitrap Exploris 480, and we will further evaluate this workflow on other platforms, and if verified by future work, this biologically relevant ion fragmentation workflow could be routinely used in many studies to improve MS/MS identification capacities.We have recently identified a pool of intracellular β1 adrenergic receptors (β1ARs) at the sarcoplasmic reticulum (SR) crucial for cardiac function. Here, we aim to characterize the integrative control of intracellular catecholamine for subcellular β1AR signaling and cardiac function. Using anchored Förster resonance energy transfer (FRET) biosensors and transgenic mice, we determined the regulation of compartmentalized β1AR-PKA signaling at the SR and plasma membrane (PM) microdomains by organic cation transporter 3 (OCT3) and monoamine oxidase A (MAO-A), two critical modulators of catecholamine uptake and homeostasis. Additionally, we examined local PKA substrate phosphorylation and excitation-contraction coupling in cardiomyocyte. Cardiac-specific deletion of MAO-A (MAO-A-CKO) elevates catecholamines and cAMP levels in the myocardium, baseline cardiac function, and adrenergic responses. Both MAO-A deletion and inhibitor (MAOi) selectively enhance the local β1AR-PKA activity at the SR but not PM, and augment phosphorylation of phospholamban, Ca2+ cycling, and myocyte contractile response. Overexpression of MAO-A suppresses the SR-β1AR-PKA activity and PKA phosphorylation. However, deletion or inhibition of OCT3 by corticosterone prevents the effects induced by MAOi and MAO-A deletion in cardiomyocytes. Deletion or inhibition of OCT3 also negates the effects of MAOi and MAO-A deficiency in cardiac function and adrenergic responses in vivo. Our data show that MAO-A and OCT3 act in concert to fine-tune the intracellular SR-β1AR-PKA signaling and cardiac fight-or-flight response. We reveal a drug contraindication between anti-inflammatory corticosterone and anti-depressant MAOi in modulating adrenergic regulation in the heart, providing novel perspectives of these drugs with cardiac implications.
Different gene expression between male and female bovine embryos leads to metabolic differences.

We used UHPLC-MS/MS to identify sex metabolite biomarkers in embryo culture medium (CM).

Embryos were produced in vitro under highly variable conditions, i.e., fertilized with 7 bulls, two breeds, and cultured with BSA or BSA + serum until Day-6. On Day-6, embryos were cultured individually for 24h. CM of Day-7 embryos (86 female and 81 male) was collected, and Day-6 and Day-7 embryonic stages recorded.

A study by sample subsets with fixed factors (culture, bull breed, and Day-6 and Day-7 stages) tentatively identified 31 differentially accumulated metabolites through 182 subsets. Day-6 and Day-7 stage together affected 13 and 11 metabolites respectively, while 19 metabolites were affected by one or another stage and/or day. Culture supplements and individual bull changed 19 and 15 metabolites, respectively. Single bull exerted the highest influence (20 metabolites with the significantly highest p values).
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