Inhibition of Xanthomonas fragariae, Causative Agent of Angular Leaf Spot of Strawberry, through Iron Deprivation.

In commercial production settings, few options exist to prevent or treat angular leaf spot (ALS) of strawberry, a disease of economic importance and caused by the bacterial pathogen Xanthomonas fragariae. In the process of isolating and identifying X. fragariae bacteria from symptomatic plants, we observed growth inhibition of X. fragariae by bacterial isolates from the same leaf macerates. Identified as species of Pseudomonas and Rhizobium, these isolates were confirmed to suppress growth of X. fragariae in agar overlay plates and in microtiter plate cultures, as did our reference strain Pseudomonas putida KT2440. Screening of a transposon mutant library of KT2440 revealed that disruption of the biosynthetic pathway for the siderophore pyoverdine resulted in complete loss of X. fragariae antagonism, suggesting iron competition as a mode of action. Antagonism could be replicated on plate and in culture by addition of purified pyoverdine or by addition of the chelating agents tannic acid and dipyridyl, while supplementing the medium with iron negated the inhibitory effects of pyoverdine, tannic acid and dipyridyl. When co-inoculated with tannic acid onto strawberry plants, X. fragariae's ability to cause foliar symptoms was greatly reduced, suggesting a possible opportunity for iron-based management of ALS. We discuss our findings in the context of 'nutritional immunity,' the idea that plant hosts restrict pathogen access to iron, either directly, or indirectly through their associated microbiota

Host response during unresolved urinary tract infection alters female mammary tissue homeostasis through collagen deposition and TIMP1

Exposure to pathogens throughout a lifetime influences immunity and organ function. Here, we explore how the systemic host-response to bacterial urinary tract infection (UTI) induces tissue-specific alterations to the mammary gland. Utilizing a combination of histological tissue analysis, single cell transcriptomics, and flow cytometry, we identify that mammary tissue from UTI-bearing mice displays collagen deposition, enlarged ductal structures, ductal hyperplasia with atypical epithelial transcriptomes and altered immune composition. Bacterial cells are absent in the mammary tissue and blood of UTI-bearing mice, therefore, alterations to the distal mammary tissue are mediated by the systemic host response to local infection. Furthermore, broad spectrum antibiotic treatment resolves the infection and restores mammary cellular and tissue homeostasis. Systemically, unresolved UTI correlates with increased plasma levels of the metalloproteinase inhibitor, TIMP1, which controls extracellular matrix remodeling and neutrophil function. Treatment of nulliparous and post-lactation UTI-bearing female mice with a TIMP1 neutralizing antibody, restores mammary tissue normal homeostasis, thus providing evidence for a link between the systemic host response during UTI and mammary gland alterations

Analysis of Lepilemur edwardsi Seasonal Acoustic Activity

Passive acoustic monitoring (PAM) has historically been used as a noninvasive technique to study biodiversity, activity patterns, distribution, and species dispersal. The use of acoustic data has expanded in ecological research; however, studies focusing on primates are underrepresented, especially for nocturnal species. This study seeks to increase the understanding of acoustic activity in the nocturnal Milne-Edwards\u27 sportive lemur (Lepilemur edwardsi). This species is endangered and endemic to the lowland dry forests in northwest Madagascar. Specifically, this study aims to describe the daily and annual acoustic activity patterns of the L. edwardsi. The PAM survey was conducted using AudioMoth recorders in the Mariarano Forest at 20 sites, with a recording being taken one minute every hour for a year. L. edwardsi presence and absence data were obtained from the recordings by using semi-automated analysis tools from ARBIMON (Rainforest Connection®). The results show that the parametric coefficients, Month and Hour, have an effect on the vocal activity of L. edwardsi with the various locations of the sites being taken into account. The probability of recording a L. edwardsi presence increases as the year progresses. Conversely, the probability of a L. edwardsi presence being recorded increases as the night progresses until midnight and decreases until the early morning hours. These results suggest that acoustic communication is important in L. edwardsi offspring rearing and parental investment. In addition, this study demonstrates the great potential of PAM in monitoring endangered and elusive species

Drug-repurposing approaches to target bacterial respiratory complexes

Antimicrobial resistance is a global health problem that resulted in an estimated 1.2 million deaths in 2019. It is predicted that antimicrobial resistance will result in an estimated 10 million deaths annually by 2050 if urgent action is not taken (Pulingam et al. 2022). This has fuelled various research aimed at discovering new antimicrobials to combat this issue. Within the past few decades, a myriad of drug targets has surfaced such as cytochrome bd. Cytochrome bd is an oxidoreductase that is found only in prokaryotes and archaea and is a promising drug candidate for numerous reasons. Cytochrome bd has been proven to increase virulence in some of the most pathogenic strains of bacteria such as Mycobacterium species, as deletion of the enzyme causes severe attenuation. The oxidase is also highly expressed and induced by the innate immune response during infection. High resolution structures of cytochrome bd from various organisms have been published which means that the oxidase can be studied computationally. These characteristics make cytochrome bd an attractive drug candidate and therefore forms the basis of this study. Lambda-red mutagenesis was previously employed by the host lab to create E. coli mutant strains that express only a single respiratory oxidase (i.e., cytochrome bd-I or bo′) and numerous attempts were made to engineer an E. coli 'cytochrome bd-II only' strain but these were unsuccessful. The current work aimed to characterise E. coli WT and respiratory mutant strains and develop an assay to measure oxygen consumption activity in membranes of these strains. Plasmid-based approaches were used to express E. coli cytochrome bd-I and cytochrome bd-II in an E. coli 'EcoM4' strain that lacked all respiratory oxidases. The strains were successfully engineered with a CO difference spectral analysis showing peaks at 640 nm. Further genetic work introduced amino acid mutations to the quinol site of E. coli cytochrome bd-I to perform future studies on antibiotic resistance. Of five amino acids that were mutated, only strains harbouring mutations in residues F269A and L253A were able to grow. Interestingly, spectral analysis revealed a distinctive peak at 640 nm for strain harbouring F269A mutation which is representative of the assembly of cytochrome bd-I. However, strain harbouring L253A mutant did not have any peaks that suggested the assembly of cytochrome bd-I. In this project both in silico and in vitro microbiological approaches were used to search for novel inhibitors of cytochrome bd. Initially, reductive approaches were undertaken to test the natural compounds madecassic acid 1 and 2-hydroxy-1,4-naphthoquinone for inhibition of E. coli cytochrome bd-I and bo′. Both compounds were inhibitory to E. coli cytochrome bd-I and bo′ but madecassic acid 1 exhibited significantly lower IC50 of 23.62 ± 7.9 µg/mL (46.80 ± 15.7 µM) and 1019 ± 260 µg/mL (6443 ± 1644 µM), respectively. Three derivatives of madecassic acid 1 were further tested to assess whether derivatisation could improve binding of the compound to cytochrome bd-I. The addition of an acetoxy group to madecassic acid 2 and 3 have shown to improve binding to cytochrome bd-I with significantly lower IC50 of 8.8 ± 3.5 µM (5.6 ± 2.2 µg/mL) and 10.2 ± 1.1 µM (8 ± 0.9 µg/mL), respectively. A drug repurposing pipeline was set-up to screen a library of FDA-approved drugs for their ability to bind to E. coli cytochrome bd-I. Steroid compounds (ethinylestradiol, quinestrol and mestranol) were identified within the top hits, and these steroid compounds were docked to the quinol site of S. aureus CydA subunit of cytochrome bd. Preliminary oxygen consumption experiments with isolated E. coli membranes identified ethinylestradiol and quinestrol as inhibitors of E. coli cytochrome bd-I. Quinestrol was the more potent compound causing inhibition of 'bd-I only' membranes with an IC50 of 0.2 ± 0.04 µg/mL (0.5 ± 0.1 µM), so this drug was selected for further analyses. Similar oxygen consumption experiments also confirmed quinestrol as an inhibitor of MRSA USA300 cytochrome bd. Growth assays showed that quinestrol completely abolished growth of MRSA USA300 WT and single oxidase mutant strains, while the growth of E. coli strains was inhibited with low IC50 values, but complete growth inhibition could not be achieved at higher concentrations of quinestrol. Survival assays demonstrated that quinestrol was lethal to MRSA bd-only cells with a median lethal concentration (LC50) of 5.6 ± 0.3 µg/mL (13.7 ± 0.7 µM). However, viability assays showed that E. coli was completely resistant to the quinestrol. This study identified novel inhibitors of cytochrome bd which will pave the way for future studies on steroid drugs as antimicrobials

AGC Team\u27s Rock Design, Heavy Civil and PreConstruction Skills at 2020 ASC Competition

MT Tech Represented: • AGC (Associated General Contractors) Club heads to Nevada • Represents the construction industry in: heavy civil, design build, and preconstructio

Patient and Provider Communication Regarding Exercise during Pregnancy in a Rural Setting

International Journal of Exercise Science 13(3): 1228-1241, 2020. Women in rural settings are at increased risk for adverse pregnancy outcomes. One potential way to improve pregnancy outcomes in rural settings is through physical activity promotion. However, given the disparities in prenatal care, women in rural areas may not receive information from their health care provider regarding physical activity during pregnancy. Therefore, the purpose of this study was to examine patient and provider communication in a rural setting (both patients’ and providers’ perspectives) regarding physical activity during pregnancy. A mixed methods study was performed among patients and providers in an obstetrical practice in a rural setting. During early pregnancy, patients were asked questions about their current physical activity levels and intentions for physical activity during their pregnancy. During late pregnancy, patients completed a survey regarding communication from their obstetric provider about exercise during pregnancy. Providers responsible for the patients’ prenatal care were surveyed regarding communication with patients about physical activity. Seventy-one pregnant women and five providers participated. 58.2% of patients reported their provider did not discuss physical activity during pregnancy with them at all. Meanwhile, all providers (100%) reported discussing physical activity with all of their patients. Similarly, only 21.8% of patients reported their provider discussed the benefits of exercise during pregnancy, while 100% of providers reported telling their patients about the benefits of exercise during pregnancy. Our study suggests ineffective patient-provider communication regarding physical activity during pregnancy in a rural setting. Improved communication strategies could reduce disparities in health outcomes among pregnant women in rural settings

Development of highly intensified cell culture perfusion media and process with tremendous productivity potential, while having a low cell bleed requirement for maintaining an overall high yield

Process intensification leveraging perfusion offers immense opportunities for yield improvement over fed-batch processes for the production of monoclonal antibodies. In the context of continuous processing, the goal is to achieve highly intensified perfusion processes that allow substantial footprint reduction and enable flexible adaptation in new facilities. Developing a productive and efficient perfusion process requires not only the application of the “push-to-low” concept for reducing the perfusion rate requirement, but also requires in-depth mechanistic development of medium formulations in order to decrease byproduct waste generation, reduce unproductive cell growth and increase productivity. Specifically reducing the usage of cell bleed is particularly desirable for improving the overall yield, since as much as 30% of the generated product may be lost through the use of cell bleed. In this work, we share case studies of perfusion medium development studying classical components such as vitamins and salts that can be manipulated to have profound effect for controlling the cell growth and reducing the use of cell bleed. In one case, the cell bleed rate was reduced down to as low as zero, while still being able to maintain a highly viable culture. Furthermore, in some cases, significant increase in the cell specific productivity (qp) was achieved when the perfusion culture was switched to a growth suppressed mode. In one example, the qp increased from 30 pg/cell/day to as high as 115 pg/cell/day when the cell growth was arrested. This led to increased daily volumetric productivities of 3 to 5 g/L/day compared to the control of 1 g/L/day. Cell cycle analysis of the arrested culture by flow cytometry also revealed an induced state of elevated cell population in the G0/G1 phase, which is generally considered as the most productive state of the cell cycle. In order to integrate the cell growth control strategy described herein, a two stage perfusion concept is designed where the first stage focuses on rapid accumulation of cells to reach the target cell density, and the second stage switches to a slow growth, yet highly productive and viable perfusion culture

Multiple system atrophy

This is a practical guide to diagnosing and managing multiple system atrophy (MSA). We explain the newly published Movement Disorders Society Consensus Diagnostic Criteria, which include new ‘Clinically Established MSA’ and ‘Possible Prodromal MSA’ categories, hopefully reducing time to diagnosis. We then highlight the key clinical features of MSA to aid diagnosis. We include a list of MSA mimics with suggested methods of differentiation from MSA. Lastly, we discuss practical symptom management in people living with MSA, including balancing side effects, with the ultimate aim of improving quality of life

Telerehabilitation Versus Standard Care for Improving Cognitive Function and Quality of Life for Adults with Traumatic Brain Injury: a Systematic Review

ABSTRACT Traumatic brain injury (TBI) is the most significant cause of death and severe disability following major trauma within Australia. Populations at risk include young adults aged 15 to 34, older adults, and military personnel. The main form of intervention following traumatic brain injury is rehabilitation, which places a large demand on the healthcare system. Telerehabilitation involves interventions delivered via telecommunication, which can improve accessibility and reduce this burden. There have been no systematic reviews conducted on the effectiveness of telerehabilitation in treating traumatic brain injury. Purpose: To examine the effectiveness of telerehabilitation for adults with traumatic brain injury. Methods: A systematic search of Medline, Embase, the Cumulative Index of Nursing and Allied Health Literature (CINAHL), Scopus, The Cochrane Library, OTSeeker and Google Scholar was conducted. Studies were included with participants aged 18 to 64 with traumatic brain injury and receiving telerehabilitation interventions. Methodological quality was assessed using the critical appraisal tools: Critical Appraisal Skills Programme (CASP) checklist for randomised controlled trials, and McMaster Critical Review for Quantitative Studies for non-randomised studies. Results: Three randomised controlled trials, one pseudo-randomised controlled trial, one case-control trial and one pre-post case series were included in this systematic review. Critical appraisal of the included studies revealed overall methodological quality to be moderate. A range of interventions with differing parameters were used as part of telerehabilitation. Collectively, there is some consistent evidence to indicate that telerehabilitation may be equally effective as other forms of care in the delivery of cognitive and psychological interventions, in addressing memory and depressive symptoms for adults with mild to severe traumatic brain injury. However, it is unclear if it is superior to other forms of care. Conclusions: A small number of studies have investigated the effect of telerehabilitation for adults with traumatic brain injury. The current evidence base is limited due to lack of standardised intervention parameters, outcomes measures and robust sample size. Despite these limitations, telerehabilitation may offer a complementary model of care for adults with traumatic brain injury, especially in instances where traditional models of care may not be readily accessible (such as those in rural and remote areas)