Establishing Guidelines to Promote Best Practice for Common Ailments in School-Based Health Clinics

The COVID-19 pandemic exposed the gap in access to medical care of school-age children and identified a need for community-based health clinics to aid in its minimization. Evidence shows the creation of school-based health clinics (SBHCs) in public school districts provides a pathway for students to receive high-quality care in an easily accessible manner. Therefore, patient care guidelines for common ailments seen in SBHCs for a local federally qualified healthcare center (FQHC) were developed in conjunction with the FQHC goal to build and operate a SBHC located in southern Illinois. Eight guidelines were created to analyze the most common chief complaints for patient visits through the use of UptoDate, evidence-based practice available from various sources and in coordination with the regulations for public school attendance in the state of Illinois. All eight guidelines were presented to the Director of Operations for the SBHC’s Mobile Units at the FQHC and consulted medical professionals. Copies of these guidelines were provided for future use to promote project sustainability. A 5-question survey was distributed to collect further feedback surrounding the guidelines, indicating likelihood of use and effectiveness in capturing proper management of common ailments. Future expansion of this project may include all staff education to providers about implementation and use of treatment guidelines once the SBHCs are opperational. This project served as an initial step toward the development of standardized evaluation and treatment of common ailments in school-based health clinics

Lipid II overproduction allows direct assay of transpeptidase inhibition by β-lactams

Peptidoglycan is an essential crosslinked polymer that surrounds bacteria and protects them from osmotic lysis. Beta-lactam antibiotics target the final stages of peptidoglycan biosynthesis by inhibiting the transpeptidases that crosslink glycan strands to complete cell wall assembly. Characterization of transpeptidases and their inhibition by beta-lactams has been hampered by lack of access to substrate. We describe a general approach to accumulate Lipid II in bacteria and to obtain large quantities of this cell wall precursor. We demonstrate utility by isolating Staphylococcus aureus Lipid II and reconstituting the synthesis of crosslinked peptidoglycan by the essential penicillin-binding protein 2, PBP2, which catalyzes both glycan polymerization and transpeptidation. We also show that we can compare the potencies of different beta-lactams by directly monitoring transpeptidase inhibition. The methods reported here will enable a better understanding of cell wall biosynthesis and facilitate studies of next-generation transpeptidase inhibitors

The Small Separation A-Star Companion Population: First Results with CHARA/MIRC-X

We present preliminary results from our long-baseline interferometry (LBI) survey to constrain the multiplicity properties of intermediate-mass A-type stars within 80pc. Previous multiplicity studies of nearby stars exhibit orbital separation distributions well-fitted with a log-normal with peaks > 15au, increasing with primary mass. The A-star multiplicity survey of De Rosa et al. (2014), sensitive beyond 30au but incomplete below 100 au, found a log-normal peak around 390au. Radial velocity surveys of slowly-rotating, chemically peculiar Am stars identified a significant number of very close companions with periods $\leq$ 5 days, ~ 0.1au, a result similar to surveys of O- and B-type primaries. With the improved performance of LBI techniques, we can probe these close separations for normal A-type stars where other surveys are incomplete. Our initial sample consists of 27 A-type primaries with estimated masses between 1.44-2.49M$_{\odot}$ and ages 10-790Myr, which we observed with the MIRC-X instrument at the CHARA Array. We use the open source software CANDID to detect five companions, three of which are new, and derive a companion frequency of 0.19$^{+0.11}_{-0.06}$ over mass ratios 0.25-1.0 and projected separations 0.288-5.481 au. We find a probability of 10$^{-6}$ that our results are consistent with extrapolations based on previous models of the A-star companion population, over mass ratios and separations sampled. Our results show the need to explore these very close separations to inform our understanding of stellar formation and evolution processes.Comment: 14 pages, 3 figures, Accepted to the Astrophysical Journal on Nov. 2, 202

MreB filaments align along greatest principal membrane curvature to orient cell wall synthesis

MreB is essential for rod shape in many bacteria. Membrane-associated MreB filaments move around the rod circumference, helping to insert cell wall in the radial direction to reinforce rod shape. To understand how oriented MreB motion arises, we altered the shape of Bacillus subtilis. MreB motion is isotropic in round cells, and orientation is restored when rod shape is externally imposed. Stationary filaments orient within protoplasts, and purified MreB tubulates liposomes in vitro, orienting within tubes. Together, this demonstrates MreB orients along the greatest principal membrane curvature, a conclusion supported with biophysical modeling. We observed that spherical cells regenerate into rods in a local, self-reinforcing manner: rapidly propagating rods emerge from small bulges, exhibiting oriented MreB motion. We propose that the coupling of MreB filament alignment to shape-reinforcing peptidoglycan synthesis creates a locally-acting, self-organizing mechanism allowing the rapid establishment and stable maintenance of emergent rod shape

Hybridization of powdery mildew strains gives rise to pathogens on novel agricultural crop species

Throughout the history of agriculture, many new crop species (polyploids or artificial hybrids) have been introduced to diversify products or to increase yield. However, little is known about how these new crops influence the evolution of new pathogens and diseases. Triticale is an artificial hybrid of wheat and rye, and it was resistant to the fungal pathogen powdery mildew (Blumeria graminis) until 2001 (refs. 1,2,3). We sequenced and compared the genomes of 46 powdery mildew isolates covering several formae speciales. We found that B. graminis f. sp. triticale, which grows on triticale and wheat, is a hybrid between wheat powdery mildew (B. graminis f. sp. tritici) and mildew specialized on rye (B. graminis f. sp. secalis). Our data show that the hybrid of the two mildews specialized on two different hosts can infect the hybrid plant species originating from those two hosts. We conclude that hybridization between mildews specialized on different species is a mechanism of adaptation to new crops introduced by agriculture

Substrate Preferences Establish the Order of Cell Wall Assembly in <i>Staphylococcus aureus</i>

The Gram-positive bacterial cell wall is a large supramolecular structure and its assembly requires coordination of complex biosynthetic pathways. In the step that merges the two major biosynthetic pathways in Staphylococcus aureus cell wall assembly, conserved protein ligases attach wall teichoic acids to peptidoglycan, but the order of biosynthetic events is a longstanding question. Here, we use a chemical approach to define which of the possible peptidoglycan intermediates are substrates for wall-teichoic acid ligases, thereby establishing the order of cell wall assembly. We have developed a strategy to make defined glycan chain-length polymers of either un-cross-linked or cross-linked peptidoglycan, and we find that wall teichoic acid ligases cannot transfer wall teichoic acid precursors to the cross-linked substrates. A 1.9 Å crystal structure of a LytR-CpsA-Psr (LCP) family ligase in complex with a wall teichoic acid precursor defines the location of the peptidoglycan binding site as a long, narrow groove, and suggests that the basis for selectivity is steric exclusion of cross-linked peptidoglycan. Consistent with this hypothesis, we have found that chitin oligomers are good substrates for transfer, showing that LCPs do not discriminate cross-linked from un-cross-linked peptidoglycan substrates by recognizing features of the un-cross-linked stem peptide. We conclude that wall teichoic acids are coupled to un-cross-linked peptidoglycan chains at an early stage of peptidoglycan synthesis and may create marks that define the proper spacing of subsequent cross-links

Substrate Preferences Establish the Order of Cell Wall Assembly in <i>Staphylococcus aureus</i>

The Gram-positive bacterial cell wall is a large supramolecular structure and its assembly requires coordination of complex biosynthetic pathways. In the step that merges the two major biosynthetic pathways in Staphylococcus aureus cell wall assembly, conserved protein ligases attach wall teichoic acids to peptidoglycan, but the order of biosynthetic events is a longstanding question. Here, we use a chemical approach to define which of the possible peptidoglycan intermediates are substrates for wall-teichoic acid ligases, thereby establishing the order of cell wall assembly. We have developed a strategy to make defined glycan chain-length polymers of either un-cross-linked or cross-linked peptidoglycan, and we find that wall teichoic acid ligases cannot transfer wall teichoic acid precursors to the cross-linked substrates. A 1.9 Å crystal structure of a LytR-CpsA-Psr (LCP) family ligase in complex with a wall teichoic acid precursor defines the location of the peptidoglycan binding site as a long, narrow groove, and suggests that the basis for selectivity is steric exclusion of cross-linked peptidoglycan. Consistent with this hypothesis, we have found that chitin oligomers are good substrates for transfer, showing that LCPs do not discriminate cross-linked from un-cross-linked peptidoglycan substrates by recognizing features of the un-cross-linked stem peptide. We conclude that wall teichoic acids are coupled to un-cross-linked peptidoglycan chains at an early stage of peptidoglycan synthesis and may create marks that define the proper spacing of subsequent cross-links