Membrane staining and phospholipid tracking in Pseudomonas aeruginosa PAO1 using the phosphatidylcholine mimic propargyl-choline.

The use of membrane-specific dyes for fluorescent microscopy is commonplace. However, most of these reagents are non-specific and cannot track specific lipid species movement, instead often acting as non-covalent lipid-associated probes or requiring the uptake of whole lipids and acyl tails into the membrane. This issue has been solved in eukaryotic cell biology by the use of click-chemistry-liable phospholipid headgroup pulse labels.

A brief descriptive analysis of a pilot study of peer support training and supervision for Texas firefighters.

Background: Peer support has been shown to reduce the incidence and severity of mental health disorders among firefighters. This pilot project had two aims: (1) to assess training satisfaction and knowledge, self-efficacy, and skills development of Texas firefighters participating in the International Association of Fire Fighters (IAFF) Peer Support Training, and (2) to determine if newly trained peer support specialists would use monthly phone supervision.

Methods: The study included 14 Texas firefighters who enrolled in the IAFF Peer Support Training.

Effects of an indole derivative on cell proliferation, transfection, and alternative splicing in production of lentiviral vectors by transient co-transfection.

Lentiviral vectors derived from human immunodeficiency virus type I are widely used to deliver functional gene copies to mammalian cells for research and gene therapies. Post-transcriptional splicing of lentiviral vector transgene in transduced host and transfected producer cells presents barriers to widespread application of lentiviral vector-based therapies. The present study examined effects of indole derivative compound IDC16 on splicing of lentiviral vector transcripts in producer cells and corresponding yield of infectious lentiviral vectors.

Transfer Printing of Ordered Plasmonic Nanoparticles at Hard and Soft Interfaces with Increased Fidelity and Biocompatibility Supports a Surface Lattice Resonance.

Transfer printing, the relocation of structures assembled on one surface to a different substrate by adjusting adhesive forces at the surface-substrate interface, is widely used to print electronic circuits on biological substrates like human skin and plant leaves. The fidelity of original structures must be preserved to maintain the functionality of transfer-printed circuits. This work developed new biocompatible methods to transfer a nanoscale square lattice of plasmon resonant nanoparticles from a lithographed surface onto leaf and glass substrates.

The Relationship Between State Level Policy, Insurance and Health Care Engagement for LGBTQ+ Foster Alumni.

Minority stress theory posits that external environmental factors such as state level policies around equity and inclusion of LGBTQ+ people can have a significant impact on the health and wellness of those identifying as part of the larger LGBTQ+ community, as well as on their decisions to seek needed physical or mental health care. This secondary data analysis explored the relationship between state level policies related to LGBTQ+ equity and inclusion and physical and mental health care engagement for foster care alumni. Using data from the Jim Casey Opportunity Passport Survey ( = 2,420), the research team conducted a longitudinal analysis of youth's engagement with healthcare professional as needed physical and mental health care, using sociodemographic characteristics of the respondents and state level policies, such as Medicaid expansion and state level protections for LGBTQ+ citizens as predictors.

Structural basis of peptidoglycan synthesis by E. coli RodA-PBP2 complex.

Peptidoglycan (PG) is an essential structural component of the bacterial cell wall that is synthetized during cell division and elongation. PG forms an extracellular polymer crucial for cellular viability, the synthesis of which is the target of many antibiotics. PG assembly requires a glycosyltransferase (GT) to generate a glycan polymer using a Lipid II substrate, which is then crosslinked to the existing PG via a transpeptidase (TP) reaction.

Activator-induced conformational changes regulate division-associated peptidoglycan amidases.

AmiA and AmiB are peptidoglycan-hydrolyzing enzymes from  that are required to break the peptidoglycan layer during bacterial cell division and maintain integrity of the cell envelope. In vivo, the activity of AmiA and AmiB is tightly controlled through their interactions with the membrane-bound FtsEX-EnvC complex. Activation of AmiA and AmiB requires access to a groove in the amidase-activating LytM domain of EnvC which is gated by ATP-driven conformational changes in FtsEX-EnvC complex.

Genetic dissection of triplicated chromosome 21 orthologs yields varying skeletal traits in Down syndrome model mice.

Down syndrome (DS) phenotypes result from triplicated genes, but the effects of three copy genes are not well known. A mouse mapping panel genetically dissecting human chromosome 21 (Hsa21) syntenic regions was used to investigate the contributions and interactions of triplicated Hsa21 orthologous genes on mouse chromosome 16 (Mmu16) on skeletal phenotypes. Skeletal structure and mechanical properties were assessed in femurs of male and female Dp9Tyb, Dp2Tyb, Dp3Tyb, Dp4Tyb, Dp5Tyb, Dp6Tyb, Ts1Rhr and Dp1Tyb;Dyrk1a+/+/- mice.

A conserved zinc-binding site in PBP2 required for elongasome-directed bacterial cell shape.

is a gram-negative bacterial pathogen that causes challenging nosocomial infections. β-lactam targeting of penicillin-binding protein (PBP)-mediated cell wall peptidoglycan (PG) formation is a well-established antimicrobial strategy. Exposure to carbapenems or zinc (Zn)-deprived growth conditions leads to a rod-to-sphere morphological transition in , an effect resembling that caused by deficiency in the RodA-PBP2 PG synthesis complex required for cell wall elongation.

Thoracolumbar Spinal Stabilization with Three Dimensional-Printed Drill Guides and Pre-Contoured Polyaxial Bone Plates.

Objective:  The aim of this study was to report new preoperative and intraoperative techniques performed for canine thoracic or lumbar spine kyphosis stabilization using three-dimensional-printed patient-specific drill guides, polyaxial titanium bone plates and drill stops, and to determine the accuracy of screw placement using these techniques.

Study Design:  Retrospective study, five client-owned dogs.

Results:  Three-dimensional-printed patient-specific drill guides and drill stops allowed safe drilling and screw placement in all of the cases, with (i) 84% of the screws graded as I (ideal placement) and 16% as IIa, IIIa or IIIb according to the modified Zdichavsky classification (partial penetration of medial pedicle wall, partial penetration of lateral pedicle wall and full penetration of lateral pedicle wall respectively), (ii) mean mediolateral deviation of ± 4.

Associations of Cardiometabolic Multimorbidity With All-Cause and Coronary Heart Disease Mortality Among Black Adults in the Jackson Heart Study.

Importance: A combination of diabetes, coronary heart disease (CHD), and stroke has multiplicative all-cause mortality risk compared with any individual morbidity in White populations, but there is a lack of studies in Black populations in the US.

Objective: To examine the association of cardiometabolic multimorbidity (diabetes, stroke, and CHD) individually and collectively with all-cause and CHD mortality.

Design, Setting, And Participants: This cohort study included Black adults in the Jackson Heart Study followed over a median of 15 years.

Investigating the effect of bacteriophages on bacterial FtsZ localisation.

is one of the most common Gram-negative pathogens and is responsible for infection leading to neonatal meningitis and sepsis. The FtsZ protein is a bacterial tubulin homolog required for cell division in most species, including . Several agents that block cell division have been shown to mislocalise FtsZ, including the bacteriophage λ-encoded Kil peptide, resulting in defective cell division and a filamentous phenotype, making FtsZ an attractive target for antimicrobials.

A 3-Dimensional Printed Patient-Specific Surgical Guide to Facilitate Transsphenoidal Hypophysectomy in Dogs.

Objective: Hypophysectomy in dogs is a difficult surgery that requires specific learning and training. We aimed to evaluate the accuracy of a 3-dimensional printed patient-specific surgical guide to facilitate choosing the entry point in the basisphenoid bone before approaching the during transsphenoidal hypophysectomy in dogs.

Methods: Two canine cadavers and 8 dogs undergoing transsphenoidal hypophysectomy for Cushing's disease treatment, involving design and fabrication of a 3-dimensional printed guide.

Plant peptidoglycan precursor biosynthesis: Conservation between moss chloroplasts and Gram-negative bacteria.

Accumulating evidence suggests that peptidoglycan, consistent with a bacterial cell wall, is synthesized around the chloroplasts of many photosynthetic eukaryotes, from glaucophyte algae to early-diverging land plants including pteridophyte ferns, but the biosynthetic pathway has not been demonstrated. Here, we employed mass spectrometry and enzymology in a two-fold approach to characterize the synthesis of peptidoglycan in chloroplasts of the moss Physcomitrium (Physcomitrella) patens. To drive the accumulation of peptidoglycan pathway intermediates, P.

Structural basis of lipopolysaccharide maturation by the O-antigen ligase.

The outer membrane of Gram-negative bacteria has an external leaflet that is largely composed of lipopolysaccharide, which provides a selective permeation barrier, particularly against antimicrobials. The final and crucial step in the biosynthesis of lipopolysaccharide is the addition of a species-dependent O-antigen to the lipid A core oligosaccharide, which is catalysed by the O-antigen ligase WaaL. Here we present structures of WaaL from Cupriavidus metallidurans, both in the apo state and in complex with its lipid carrier undecaprenyl pyrophosphate, determined by single-particle cryo-electron microscopy.

Characterization of Secondary Structure and Thermal Stability by Biophysical Methods of the D-alanyl,D-alanine Ligase B Protein from .

Background: Peptidoglycan (PG) is a key structural component of the bacterial cell wall and interruption of its biosynthesis is a validated target for antimicrobials. Of the enzymes involved in PG biosynthesis, D-alanyl,D-alanine ligase B (DdlB) is responsible for the condensation of two alanines, forming D-Ala-D-Ala, which is required for subsequent extracellular transpeptidase crosslinking of the mature peptidoglycan polymer.

Objective: We aimed at the biophysical characterization of recombinant Escherichia coli DdlB (EcDdlB), considering parameters of melting temperature (T), calorimetry and Van't Hoff enthalpy changes of denaturation ( ΔH and ΔH ), as well as characterization of elements of secondary structure at three different pHs.

A Dynamic Network of Proteins Facilitate Cell Envelope Biogenesis in Gram-Negative Bacteria.

Bacteria must maintain the ability to modify and repair the peptidoglycan layer without jeopardising its essential functions in cell shape, cellular integrity and intermolecular interactions. A range of new experimental techniques is bringing an advanced understanding of how bacteria regulate and achieve peptidoglycan synthesis, particularly in respect of the central role played by complexes of Sporulation, Elongation or Division (SEDs) and class B penicillin-binding proteins required for cell division, growth and shape. In this review we highlight relationships implicated by a bioinformatic approach between the outer membrane, cytoskeletal components, periplasmic control proteins, and cell elongation/division proteins to provide further perspective on the interactions of these cell division, growth and shape complexes.

Synthetic Sansanmycin Analogues as Potent Translocase I Inhibitors.

Herein, we report the design and synthesis of inhibitors of () phospho-MurNAc-pentapeptide translocase I (MurX), the first membrane-associated step of peptidoglycan synthesis, leveraging the privileged structure of the sansanmycin family of uridylpeptide natural products. A number of analogues bearing hydrophobic amide modifications to the pseudo-peptidic end of the natural product scaffold were generated that exhibited nanomolar inhibitory activity against MurX and potent activity against . We show that a lead analogue bearing an appended neopentylamide moiety possesses rapid antimycobacterial effects with a profile similar to the frontline tuberculosis drug isoniazid.

The Pneumococcal Divisome: Dynamic Control of Cell Division.

Cell division in (pneumococcus) is performed and regulated by a protein complex consisting of at least 14 different protein elements; known as the divisome. Recent findings have advanced our understanding of the molecular events surrounding this process and have provided new understanding of the mechanisms that occur during the division of pneumococcus. This review will provide an overview of the key protein complexes and how they are involved in cell division.

Wastewater Surveillance for SARS-CoV-2 on College Campuses: Initial Efforts, Lessons Learned, and Research Needs.

Wastewater surveillance for the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an emerging approach to help identify the risk of a coronavirus disease (COVID-19) outbreak. This tool can contribute to public health surveillance at both community (wastewater treatment system) and institutional (e.g.

A molecular link between cell wall biosynthesis, translation fidelity, and stringent response in .

Survival in the human host requires bacteria to respond to unfavorable conditions. In the important Gram-positive pathogen , cell wall biosynthesis proteins MurM and MurN are tRNA-dependent amino acyl transferases which lead to the production of branched muropeptides. We demonstrate that wild-type cells experience optimal growth under mildly acidic stressed conditions, but Δ strain displays growth arrest and extensive lysis.

Structure-based modeling and dynamics of MurM, a Streptococcus pneumoniae penicillin resistance determinant present at the cytoplasmic membrane.

Branched Lipid II, required for the formation of indirectly crosslinked peptidoglycan, is generated by MurM, a protein essential for high-level penicillin resistance in the human pathogen Streptococcus pneumoniae. We have solved the X-ray crystal structure of Staphylococcus aureus FemX, an isofunctional homolog, and have used this as a template to generate a MurM homology model. Using this model, we perform molecular docking and molecular dynamics to examine the interaction of MurM with the phospholipid bilayer and the membrane-embedded Lipid II substrate.

Correlation of SARS-CoV-2 RNA in wastewater with COVID-19 disease burden in sewersheds.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which causes the coronavirus disease (COVID-19), is shed in feces and the viral ribonucleic acid (RNA) is detectable in wastewater. A nine-week wastewater epidemiology study of ten wastewater facilities, serving 39% of the state of Utah or 1.26 M individuals was conducted in April and May of 2020.

Wastewater surveillance for SARS-CoV-2 on college campuses: Initial efforts, lessons learned and research needs.

Background: Wastewater surveillance for SARS-CoV-2 is an emerging approach to help identify the risk of a COVID-19 outbreak. This tool can contribute to public health surveillance at both community (wastewater treatment system) and institutional (e.g.