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Vitamin K epoxide reductases (VKOR) constitute a major family of integral membrane thiol oxidoreductases. In humans, VKOR sustains blood coagulation and bone mineralization through the vitamin K cycle. Previous chemical models assumed that the catalysis of human VKOR (hVKOR) starts from a fully reduced active site. This state, however, constitutes only a minor cellular fraction (5.6%). RMC-6236 clinical trial Thus, the mechanism whereby hVKOR catalysis is carried out in the cellular environment remains largely unknown. Here we use quantitative mass spectrometry (MS) and electrophoretic mobility analyses to show that KO likely forms a covalent complex with a cysteine mutant mimicking hVKOR in a partially oxidized state. Trapping of this potential reaction intermediate suggests that the partially oxidized state is catalytically active in cells. To investigate this activity, we analyze the correlation between the cellular activity and the cellular cysteine status of hVKOR. We find that the partially oxidized hVKOR has considerably lower activity than hVKOR with a fully reduced active site. Although there are more partially oxidized hVKOR than fully reduced hVKOR in cells, these two reactive states contribute about equally to the overall hVKOR activity, and hVKOR catalysis can initiate from either of these states. Overall, the combination of MS quantification and biochemical analyses reveal the catalytic mechanism of this integral membrane enzyme in a cellular environment. Furthermore, these results implicate how hVKOR is inhibited by warfarin, one of the most commonly prescribed drugs.Determination of cellular ATP levels, a key indicator of metabolic status, is essential for the quantitative analysis of metabolism. The biciliate green alga Chlamydomonas reinhardtii is an excellent experimental organism to study ATP production pathways, including photosynthesis and respiration, particularly because it can be cultured either photoautotrophically or heterotrophically. Additionally, its cellular ATP concentration, [ATP], is reflected in the beating of its cilia. However, the methods currently used for quantifying the cellular ATP levels are time-consuming or invasive. In this study, we established a rapid method for estimating cytosolic [ATP] from the ciliary beating frequency (CBF) in C. reinhardtii Using an improved method of motility reactivation in demembranated cell models, we obtained calibration curves for [ATP]-CBF over a physiological range of ATP concentrations. These curves allowed rapid estimation of the cytosolic [ATP] in live wild-type cells to be ~2.0 mM in the light and ~1.5 mM in the dark values comparable to those obtained by other methods. Furthermore, we used this method to assess the effects of genetic mutations or inhibitors of photosynthesis or respiration quantitatively and non-invasively. This sensor-free method is a convenient tool for quickly estimating cytosolic [ATP] and studying the mechanism of ATP production in C. reinhardtii or other ciliated organisms.

Bedside delivery of discharge medications improves caregiver understanding and experience. Less is known about its impact on medication adherence. We aimed to improve antimicrobial adherence by increasing on-time first home doses for patients discharged from the pediatric hospital medicine service from 33% to 80% over 1 year via creation of a discharge medication delivery and counseling "Meds to Beds" (M2B) program.

Using sequential plan-do-study-act cycles, an interprofessional workgroup implemented M2B on select pediatric hospital medicine units at our quaternary children's hospital from October 2017 through December 2018. Scripted telephone surveys were conducted with caregivers of patients prescribed antimicrobial agents at discharge. The primary outcome measure was on-time administration of the first home antimicrobial dose, defined as a dose given within the time of the inpatient dose equivalent plus 25%. Process measures primarily assessed caregiver report of barriers to adherence. Run charts, statbefore discharge. Importantly, caregivers reported reduced barriers to medication adherence.In recent years, Lake Baikal has undergone significant changes in the composition of coastal communities associated with the increasing anthropogenic influence and global climate changes. In this context, we carried out metagenomic sequencing of the DNA viral community of an integral near-bottom water sample from the littoral zone of the lake.Here, we report the complete genome sequence of Adlercreutzia sp. strain 8CFCBH1, which was isolated from a Japanese fecal sample. The genome analysis revealed that the 8CFCBH1 strain potentially produces (S)-equol.The antimony-oxidizing Stenotrophomonas sp. strain SbOxS2 was isolated from stibnite mine tailing soil. The draft genome sequence of strain SbOxS2 comprises 4.76 Mbp with 4,211 predicted protein-coding sequences. This genome will provide useful information for characterizing the molecular mechanisms associated with heavy metal resistance within the genus Stenotrophomonas.Pseudomonas sp. strain SbOxS1 was isolated from stibnite mine tailing soil for its ability to oxidize antimonite. We present a draft genome sequence of strain SbOxS1, which contains 6,484 predicted protein-coding sequences. This genome information extends our understanding of the physiological versatility of antimony-transforming microorganisms.The Gram-negative bacterium Vibrio nigripulchritudo is an important shrimp pathogen. Here, we present the genome sequence of Vibrio nigripulchritudo TUMSAT-TG-2018, which was isolated from a diseased Pacific white shrimp (Litopenaeus vannamei). The assembly totaled 6.8 Mbp, consisting of two chromosomes and four plasmids.Phaeovibrio sulfidiphilus was reported to be a divergent member of the purple photosynthetic bacteria with limited ability to metabolize organic compounds. Whole-genome-based analysis shows that it is indeed only distantly related to freshwater species of Rhodospirillaceae Unexpectedly, the genome contains unique gene clusters for potential respiratory nitrate reduction and anaerobic glycerol metabolism.Here, we report the draft genome sequence of Lactobacillus rhamnosus NCB 441, which was isolated from pickled white cheese samples gathered at Farafra Oasis in New Valley Governorate, Egypt. The genome size is 2,969,245 bp with a G+C content of 46.7%.The complete genome of Salmonella enterica subsp. enterica serovar Kottbus strain Kharkiv (serogroup C2-C3), which was isolated from a commercial pork production facility in Kharkiv, Ukraine, was assembled using long-read Nanopore sequences. A single circular contig (4,799,045 bp) comprised a complete chromosome encoding antibiotic resistance, highlighting the risk of cross-species livestock and human infection.Aspergillus flavus is an agriculturally and medically important filamentous fungus that produces mycotoxins, including aflatoxins, which are potent carcinogens. Here, we generated short- and long-read transcript sequence data from the growth of A. flavus strain NRRL 3357 under both typical and stress conditions to produce a new annotation of its genome.Aphanizomenon flos-aquae is a significant harmful algal bloom-forming cyanobacterial species. Here, we report the draft genome for a strain of A. flos-aquae (Clear-A1) from a harmful algal bloom enrichment culture. This metagenome-assembled genome (MAG) sequence comprises 4,452,466 bp in 60 contigs with a GC content of 37.1%.Dolichospermum circinale (formerly Anabaena circinale) is a significant harmful algal bloom species. We report the draft metagenome-assembled genome (MAG) for a strain of D. circinale (Clear-D4) obtained from an enrichment culture. The genome sequence comprises 5,029,933 bp in 560 contigs with a GC content of 37%.We report the metagenome-assembled genome sequence of a Vulcanococcus sp. binned from a cyanobacterial enrichment culture. The genome contains 39 contigs comprising 2.96 Mbp and is estimated as 100% complete, with a GC content of 63.9% and 3,261 predicted coding genes.Here, we report the metagenome-assembled genome sequence of a Rhodobacteraceae bacterium strain, Clear-D3, that was reconstructed from a cyanobacterial enrichment from a eutrophic lake. The draft genome sequence shows evidence of an anoxygenic photoautotrophic lifestyle. Other potential capabilities include aerobic heterotrophy, flagellar motility, chemotaxis, and utilization of complex C-P compounds.Metagenomic sequencing of a Dolichospermum circinale enrichment culture resulted in the assembly of several cocultured metagenome-assembled genomes (MAGs). One MAG was affiliated with the class Kapabacteriales and included 5,724,991 bp in 127 contigs with a GC content of 48.4%.Pneumonic pasteurellosis, caused by Pasteurella multocida, is a common respiratory infection of ruminants that has major economic and welfare implications throughout the world. Here, we report the annotated genome sequences of seven pathogenic strains of P. multocida that were isolated from cattle in the United Kingdom.Land use is a major factor contributing to the differences in soil microbial assemblages. Despite the importance of microbial communities on crop health and productivity, a knowledge gap exists on the effects of land use change on microbial functions in the rhizosphere. This data set presents the metagenomic data from two maize fields in South Africa with different agricultural histories. It provides an opportunity for modeling microbes with beneficial functions that could enhance crop productivity.Methicillin-resistant Staphylococcus aureus sequence type 72 (ST72) is prevalent in South Korea and has shown resistance to multiple antimicrobials. ST72 isolates display different levels of resistance to the antistaphylococcal lysostaphin. Draft genome sequencing of ST72 human isolates exhibiting lysostaphin resistance or susceptibility was performed to better understand the mechanism of lysostaphin resistance using subtractive genomics.The circular extrachromosomal ribosomal DNA (rDNA) element of Naegleria fowleri strain LEE was molecularly cloned and fully sequenced. The element comprises 15,786 bp and contains a single copy of the organism's rDNA cistron. The nonribosomal sequence contains five potential open reading frames, two large direct repeat sequences, and numerous smaller repeated-sequence regions.We present the complete genomes of four Brucella suis biovar 2 isolates that were obtained from wild boars in Switzerland in 2008 and 2009. Genomes were sequenced with PacBio technology, contained two chromosomes each, had a genome size of 3.3 Mbp, and contained more than 3,225 genes per genome.Here, we present the complete genome sequence of Marinobacterium sp. strain LSUCC0821, isolated from the coastal Gulf of Mexico with artificial seawater using high-throughput dilution-to-extinction (DTE) cultivation. The 2.36-Mbp circularized genome sequence has 2,231 predicted genes, a 91.5% coding density, and a GC content of 47.8%.Cotton is an important cash crop for both the Indian economy and rural livelihoods. In the present study, metagenomic analysis is used to characterize microbial diversity in cotton rhizosphere soil from the Alwar district, located in the semiarid northeast region of the state of Rajasthan in India.