David William Koenig

Vaginal Microbiome and Epithelial Gene Array in Post-Menopausal Women with Moderate to Severe Dryness

After menopause, many women experience vaginal dryness and atrophy of tissue, often attributed to the loss of estrogen. An understudied aspect of vaginal health in women who experience dryness due to atrophy is the role of the resident microbes. It is known that the microbiota has an important role in healthy vaginal homeostasis, including maintaining the pH balance and excluding pathogens. The objectives of this study were twofold: first to identify the microbiome of post-menopausal women with and without vaginal dryness and symptoms of atrophy; and secondly to examine any differences in epithelial gene expression associated with atrophy. The vaginal microbiome of 32 post-menopausal women was profiled using Illumina sequencing of the V6 region of the 16S rRNA gene. Sixteen subjects were selected for follow-up sampling every two weeks for 10 weeks. In addition, 10 epithelial RNA samples (6 healthy and 4 experiencing vaginal dryness) were acquired for gene expression analysis by Affymetrix Human Gene array. The microbiota abundance profiles were relatively stable over 10 weeks compared to previously published data on premenopausal women. There was an inverse correlation between Lactobacillus ratio and dryness and an increased bacterial diversity in women experiencing moderate to severe vaginal dryness. In healthy participants, Lactobacillus iners and L. crispatus were generally the most abundant, countering the long-held view that lactobacilli are absent or depleted in menopause. Vaginal dryness and atrophy were associated with down-regulation of human genes involved in maintenance of epithelial structure and barrier function, while those associated with inflammation were up-regulated consistent with the adverse clinical presentation.

Highly Potent Inhibitors of Quorum Sensing in Staphylococcus aureus Revealed Through a Systematic Synthetic Study of the Group-III Autoinducing Peptide

Methods to intercept bacterial quorum sensing (QS) have attracted significant attention as potential anti-infective therapies. Staphylococcus aureus is a major human pathogen that utilizes autoinducing peptide (AIP) signals to mediate QS and thereby regulate virulence. S. aureus strains are categorized into four groups (I-IV) according to their AIP signal and cognate extracellular receptor, AgrC. Each group is associated with a certain disease profile, and S. aureus group-III strains are responsible for toxic shock syndrome and have been underestimated in other infections to date. A limited set of non-native AIP analogs have been shown to inhibit AgrC receptors; such compounds represent promising tools to study QS pathways in S. aureus . We seek to expand this set of chemical probes and report herein the first design, synthesis, and biological testing of AIP-III mimetics. A set of non-native peptides was identified that can inhibit all four of the AgrC receptors (I-IV) with picomolar IC50 values in reporter strains. These analogs also blocked hemolysis by wild-type S. aureus group I-IV strains-a virulence trait under the control of QS-at picomolar concentrations. Moreover, four of the lead AgrC inhibitors were capable of attenuating the production of toxic shock syndrome toxin-1 (also under the control of QS) by over 80% at nanomolar concentrations in a wild-type S. aureus group-III strain. These peptides represent, to our knowledge, the most potent synthetic inhibitors of QS in S. aureus known, and constitute new and readily accessible chemical tools for the study of the AgrC system and virulence in this deadly pathogen.

A systems biology approach investigating the effect of probiotics on the vaginal microbiome and host responses in a double blind, placebo-controlled clinical trial of post-menopausal women.

A lactobacilli dominated microbiota in most pre and post-menopausal women is an indicator of vaginal health. The objective of this double blinded, placebo-controlled crossover study was to evaluate in 14 post-menopausal women with an intermediate Nugent score, the effect of 3 days of vaginal administration of probiotic L. rhamnosus GR-1 and L. reuteri RC-14 (2.5×109 CFU each) on the microbiota and host response. The probiotic treatment did not result in an improved Nugent score when compared to when placebo. Analysis using 16S rRNA sequencing and metabolomics profiling revealed that the relative abundance of Lactobacillus was increased following probiotic administration as compared to placebo, which was weakly associated with an increase in lactate levels. A decrease in Atopobium was also observed. Analysis of host responses by microarray showed the probiotics had an immune-modulatory response including effects on pattern recognition receptors such as TLR2 while also affecting epithelial barrier function. This is the first study to use an interactomic approach for the study of vaginal probiotic administration in post-menopausal women. It shows that in some cases multifaceted approaches are required to detect the subtle molecular changes induced by the host to instillation of probiotic strains. Trial Registration ClinicalTrials.gov NCT02139839

Decreased activity of commercially available disinfectants containing quaternary ammonium compounds when exposed to cotton towels

BACKGROUND Health care-associated infections (HAIs) are a significant problem in hospitals, and environmental surfaces have been implicated as a source of HAIs in the hospital environment. Furthermore, Gram-negative and Gram-positive bacteria can persist on dry environmental surfaces for as long as several months. Poorly cleaned surfaces may serve as vehicles for microbes, which may then be transferred to patients. METHODS Cotton and microfiber towels were both tested for their abilities to bind quaternary ammonium compounds (QACs). The towels were exposed to 3 commercially available disinfectants for 0.5, 30, and 180 minutes. Germicidal spray tests (GSTs) were performed for all towel eluates in accordance with the AOAC International method 961.02. Cotton towel eluates were analyzed for QAC concentration using high-performance liquid chromatography. RESULTS QAC concentrations were reduced by up to 85.3% after exposure to cotton towels, resulting in failure of the disinfectants exposed to cotton towels in 96% of the GSTs. CONCLUSION The use of cotton towels with QAC-based cleansers should be reconsidered, particularly in hospitals where effective cleaning of the patient environment is needed to reduce the risk of HAIs.

Immediate and extended effects of sodium lauryl sulphate exposure on stratum corneum natural moisturizing factor

Natural moisturizing factor (NMF) serves as the primary humectant of the stratum corneum (SC), principally comprised of hygroscopic amino acids and derivatives that absorb moisture. Barrier disruption has been shown to differentially affect the levels of specific NMF components, though the kinetics of NMF component restoration following disruption have not been examined. Here, we investigated the impact of barrier disruption caused by surfactant exposure on a subset of NMF components immediately following exposure and out to 10 days post‐exposure.

Modeling of Human Viruses on Hands and Risk of Infection in an Office Workplace Using Micro-Activity Data

Although the number of illnesses resulting from indirect viral pathogen transmission could be substantial, it is difficult to estimate the relative risks because of the wide variation and uncertainty in human behavior, variable viral concentrations on fomites, and other exposure factors. The purpose of this study was to evaluate the micro-activity approach for assessment of microbial risk by adapting a mathematical model to estimate probability of viral infection from indirect transmission. To evaluate the model, measurements of phage loading on fomites and hands collected before and after implementation of a Healthy Workplace Project intervention were used. Parameter distributions were developed from these data, as well as for micro-activity rates, contact surface areas, phage transfer efficiencies, and inactivation rates. Following the Monte Carlo simulations (n = 1,000), the estimated phage loading on hands was not significantly different from the loading of phage on hands measured in the experimental trials. The model was then used to demonstrate that the Healthy Workplace Project intervention significantly reduced risk of infection by 77% for rotavirus and rhinovirus. This is the first published study to successfully evaluate a model focused on the indirect transmission of viruses via hand contact with measured data and provide an assessment of the micro-activity approach to microbial risk evaluation.

Microbial contamination of hospital reusable cleaning towels

BACKGROUND Hospital cleaning practices are critical to the prevention of nosocomial infection transmission. To this end, cloth towels soaked in disinfectants are commonly used to clean and disinfect hospital surfaces. Cloth cleaning towels have been linked to an outbreak of Bacillus cereus and have been shown to reduce the effectiveness of commonly used quaternary ammonium disinfectants. Thus, it is important to determine whether the reuse of cloth towels increases the risk of pathogen transmission in hospitals. METHODS The goal of this project was to determine the effects of laundry and cleaning practices commonly used in hospitals for washing, storage, and disinfection of cloth cleaning towels on their microbial loads. RESULTS Our results indicate that cloth towels used for cleaning hospital rooms contained high numbers of microbial contaminants. CONCLUSIONS In this case, hospital laundering practices appear insufficient to remove microbial contaminants and may even add contaminants to the towels. Furthermore, it has been previously reported that towels can interfere with the action of common hospital disinfectants. Either independently or in combination, these 2 factors may increase the risk for transmission of pathogens in hospitals. These observations indicate the need to critically reevaluate current hospital cleaning practices associated with reuse of cloth towels.

The healthy workplace project: Reduced viral exposure in an office setting

ABSTRACT Viral illnesses such as gastroenteritis and the common cold create a substantial burden in the workplace due to reduced productivity, increased absenteeism, and increased health care costs. Behaviors in the workplace contribute to the spread of human viruses via direct contact between hands, contaminated surfaces, and the mouth, eyes, and/or nose. This study assessed whether implementation of the Healthy Workplace Project (HWP) (providing hand sanitizers, disinfecting wipes, facial tissues, and use instructions) would reduce viral loads in an office setting of approximately 80 employees after seeding fomites and the hands of volunteer participants with an MS-2 phage tracer. The HWP significantly reduced viable phage detected on participants’ hands, communal fomites, and personal fomites (p ≤ .010) in office environments and presents a cost-effective method for reducing the health and economic burden associated with viral illnesses in the workplace.

Immediate and extended effects of abrasion on stratum corneum natural moisturizing factor

Natural moisturizing factor (NMF), principally comprised of hygroscopic amino acids and derivatives that absorb moisture from the surrounding environment, serves as the primary humectant of the stratum corneum (SC). Acute barrier disruption has been shown to differentially affect the concentration of NMF in the SC. This study measured the recovery kinetics of NMF after mechanical damage of the SC, which is not well understood.

In vitro prediction of in vivo skin damage associated with the wiping of dry tissue against skin

The ideal gentle cleansing product is one that effectively removes soils while minimizing damage to the skin. Thus, measuring physical abrasion caused by cleansing tissues is critical to the continued development of gentle cleansing products. Current analysis of cleansing materials for skin gentleness is time consuming and requires expensive human subject testing. This report describes the development of a rapid and inexpensive bench assay for the assessment of skin abrasion caused by wiping.