A method for culturing Gram-negative skin microbiota.

BACKGROUND: Commensal Gram-negative (CGN) microbiota have been identified on human skin by DNA sequencing; however, methods to reliably culture viable Gram-negative skin organisms have not been previously described. RESULTS: Through the use of selective antibiotics and minimal media we developed methods to culture CGN from skin swabs. We identified several previously uncharacterized CGN at the species level by optimizing growth conditions and limiting the inhibitory effects of nutrient shock, temperature, and bacterial competition, factors that may have previously limited CGN isolation from skin cultures. CONCLUSIONS: Our protocol will permit future functional studies on the influences of CGN on skin homeostasis and disease.

Reproductive Experiences of Physicians in Medical and Surgical Subspecialties.

Objective: To evaluate the reproductive experiences of physicians across gender and specialty. Patients and Methods: Between November and December 2021, we surveyed nontrainee physicians of all genders at a single quaternary institution using a modified version of an existing survey instrument. Experiences with family planning, fertility, pregnancy, and parental leave were assessed. Results: There were 422 completed responses. Respondents reported a higher prevalence of infertility as compared to the general U.S. population (26% versus 19%), with no difference in infertility or obstetrical complications by specialty. Most respondents (75%) reported stigma regarding having children in medicine, and 71% reported delaying childbearing. These trends were strongest in the subanalysis of female respondents. Forty-five percent of respondents reported that their work increased the risk for subfertility, infertility, or pregnancy complications. Surgeons were significantly more likely to report physically demanding work conditions (75% versus 30%, p < 0.001), radiation exposure (39% versus 14%, p < 0.001), and bloodborne pathogen exposure (25% versus 12%, p = 0.03) as occupational reproductive hazards. Only 55% of respondents with a pregnancy history reported ever taking parental leave. Among those who took less than the full amount offered, 63% cited concerns about falling behind educationally or professionally as significantly influencing this decision. Conclusions: These results support previous trends showing delayed childbearing and increased infertility among physicians while shedding new light on stigma associated with childbearing and parental leave. A better understanding of the reproductive experiences of physicians is critical to recruiting and retaining a skilled workforce and fostering career and life satisfaction in this profession.

Fertility preservation options for transgender and nonbinary individuals.

Transgender and nonbinary individuals are historically underserved by healthcare systems. A crucial area for improvement is fertility preservation counseling and service delivery, as gender-affirming hormone therapy and gender-affirming surgery may negatively affect future fertility. The methods available for fertility preservation depend on the patient's pubertal status and utilization of gender-affirming therapies, and counseling and delivery of these services are complex and require a multidisciplinary approach. Further research is needed to identify pertinent stakeholders in managing the care of these patients, as well as to better understand the optimal frameworks for delivering integrated and comprehensive care to this patient population. Fertility preservation is an active and exciting area of scientific discovery and offers a wealth of opportunities to improve the care of transgender and nonbinary individuals.

Impact of the COVID-19 Pandemic on Obstetricians/Gynecologists.

OBJECTIVE: To assess the impact of the coronavirus disease 2019 (COVID-19) pandemic on obstetricians/gynecologists (OB/GYNs). PARTICIPANTS AND METHODS: A 49-item survey was distributed to OB/GYNs through the websites and electronic mailing lists of professional OB/GYN organizations. The survey was open from June 22, 2020, through November 22, 2020. Of the 122 initiated surveys, 89 were completed (73.0% completion rate); 72 respondents answered at least one open-ended question and were included for qualitative analysis. RESULTS: Respondents reported policy changes, limited personal protective equipment availability, patient compliance with safety protocols and personal protective equipment use, staff shortages, and concerns about COVID-19 exposure as primary stressors related to the pandemic. Respondents felt that the pandemic had a negative professional impact on their relationships with patients and colleagues. Workplace and pandemic stressors resulted in feelings of anxiety and frustration; physical effects were also reported. Some respondents indicated that they were considering early retirement or leaving the profession as a result of the pandemic, which suggests that OB/GYNs may be at increased risk for burnout. CONCLUSION: The COVID-19 pandemic will have important long-term effects on OB/GYN well-being and workforce retention. Proactive support for OB/GYNs is needed to combat burnout and counteract workforce attrition. Implementing peer support programs that promote healthy emotional processing following adverse events may mitigate these feelings and reduce OB/GYN burnout.

Continuous positive airway pressure with deep inspiration breath hold in left-sided breast radiation therapy.

A dosimetric study to evaluate the use of continuous positive airway pressure (CPAP), with free-breathing (CPAP-FB) or with deep inspiration breath hold (DIBH-CPAP) an adjunct and alternative to DIBH to reduce heart and lung dose in the radiation therapy (RT) of breast cancer planned for left side RT with regional nodes and internal mammary. A retrospective analysis of 10 left-sided breast cancer patients whose heart or lung dose constraints were not met after RT planning based on FB or DIBH simulations and were referred for CPAP-based planning. All patients were simulated using FB, DIBH, CPAP-FB, and CPAP-DIBH. Treatment plans were calculated to cover the breast/chest wall and regional nodes using tangential field-in-field technique (FiF). Dose-volume parameters for heart, ipsilateral lung, and contralateral breast were compared using the Wilcoxon signed-rank test. For all RT plans, mean heart dose (Gy) was lower for treatment plans with CPAP: CPAP-FB (mean 3.4 vs 7.4, p = 0.001) and CPAP-DIBH (mean 2.5 vs 7.4, p = 0.006) compared to FB alone. CPAP-DIBH also significantly reduced MHD as compared to DIBH alone (mean 2.5 vs 4.3 Gy, p = 0.013). CPAP-DIBH significantly reduced mean lung dose as compared to both FB (mean 14.4 vs 20.1, p = 0.005) and DIBH alone (mean 14.4 vs 17.4, p = 0.007). Eight of 10 patients did not meet ipsilateral lung V20Gy dose constraints (≥35% of lung receiving 20 Gy) in either the free breathing or DIBH plans, whereas 8 out of 10 met lung V20Gy goal constraints (≤30% of lung receiving 20 Gy) in the CPAP-DIBH plans. Based on the outcomes of our study, CPAP could be a strategy for reducing lung and heart dose, both in patients not able to execute DIBH and as an adjunct in those not deriving sufficient dose reduction from DIBH alone.

Kerion of the pubis and vulva with bacterial superinfection: a rare occurrence.

Kerion is a severe hypersensitivity reaction to fungal infection that is rarely seen in the groin. Frequent shaving of pubic hair and religious conservatism surrounding genital hygiene are common among Bedouin women in the Negev Desert, and may predispose to kerion. This case highlights the clinical course of a 20-year-old Bedouin woman who presented with severe kerion celsi of the pubis and vulva with secondary bacterial infection. The patient was successfully treated with intravenous antibiotics, oral antifungal medication and wet topical dressings. The case outlines the risk factors and treatment for severe kerion celsi of the groin, as well as possible preventive measures that may reduce its incidence.

Subungual and ungual scabies: avoiding severe presentation in high-risk patients.

Scabies is a pruritic disorder caused by Sarcoptes scabiei var. hominis infestation of the skin. Transferred by close body contact, scabies is endemic within nursing homes and among poor and overcrowded populations. Crusted scabies is a severe form of disease, characterised by a large, thick, crusted eruption with significant mite infestation. We report a patient hospitalised with crusted scabies that had massive nail involvement. A 79-year-old female patient with multiple comorbidities and several recent prior scabies diagnoses presented with agitation and dystrophic fingernails; scabies mites were found embedded in and below the nail keratin. Aggressive treatment resulted in complete resolution, with notable improvements in mental status. Crusted scabies with nail involvement is extremely rare and may be more likely to develop from initially subclinical infestation sites. It is important to consider this potential presentation, as standard topical treatments may prove ineffective when there is deep nail involvement.

The Choroid Plexus Functions as a Niche for T-Cell Stimulation Within the Central Nervous System.

The choroid plexus (CP) compartment in the ventricles of the brain comprises fenestrated vasculature and, therefore, it is permeable to blood-borne mediators of inflammation. Here, we explored whether T-cell activation in the CP plays a role in regulating central nervous system (CNS) inflammation. We show that CD4 T cells injected into the lateral ventricles adhere to the CP, transmigrate across its epithelium, and undergo antigen-specific activation and proliferation. This process is enhanced following peripheral immune stimulation and significantly impacts the immune signaling induced by the CP. Ex vivo studies demonstrate that T-cell harboring the CP through its apical surface is a chemokine- and adhesion molecule-dependent process. We suggest that, within the CNS, the CP serves an immunological niche, which rapidly responds to peripheral inflammation and, thereby, promotes two-way T-cell trafficking that impact adaptive immunity in the CNS.

Molecular Typing of Staphylococcus aureus Isolated from Patients with Autosomal Dominant Hyper IgE Syndrome.

Autosomal dominant hyper IgE syndrome (AD-HIES) is a primary immunodeficiency caused by a loss-of-function mutation in the Signal Transducer and Activator of Transcription 3 (STAT3). This immune disorder is clinically characterized by increased susceptibility to cutaneous and sinopulmonary infections, in particular with Candida and Staphylococcus aureus. It has recently been recognized that the skin microbiome of patients with AD-HIES is altered with an overrepresentation of certain Gram-negative bacteria and Gram-positive staphylococci. However, these alterations have not been characterized at the species- and strain-level. Since S. aureus infections are influenced by strain-specific expression of virulence factors, information on colonizing strain characteristics may provide insights into host-pathogen interactions and help guide management strategies for treatment and prophylaxis. The aim of this study was to determine whether the immunodeficiency of AD-HIES selects for unique strains of colonizing S. aureus. Using multi-locus sequence typing (MLST), protein A (spa) typing, and PCR-based detection of toxin genes, we performed a detailed analysis of the S. aureus isolates (n = 13) found on the skin of twenty-one patients with AD-HIES. We found a low diversity of sequence types, and an abundance of strains that expressed methicillin resistance, Panton-Valentine leukocidin (PVL), and staphylococcal enterotoxins K and Q (SEK, SEQ). Our results indicate that patients with AD-HIES may often carry antibiotic-resistant strains that harbor key virulence factors.

CD8+ T cells produce a dialyzable antigen-specific activator of dendritic cells.

Cellular lysates from PPD+ donors have been reported to transfer tuberculin reactivity to naïve recipients, but not diphtheria reactivity, and vice versa. A historically controversial topic, the terms "transfer factor" and "DLE" were used to characterize the reactivity-transferring properties of lysates. Intrigued by these reported phenomena, we found that the cellular extract derived from antigen-specific memory CD8+ T cells induces IL-6 from antigen-matched APCs. This ultimately elicits IL-17 from bystander memory CD8+ T cells. We have identified that dialyzable peptide sequences, S100a9, and the TCR ß chain from CD8+ T cells contribute to the molecular nature of this activity. We further show that extracts from antigen-targeted T cells enhance immunity to Staphylococcus aureus and Candida albicans These effects are sensitive to immunization protocols and extraction methodology in ways that may explain past discrepancies in the reproducibility of passive cellular immunity.

Transplantation of human skin microbiota in models of atopic dermatitis.

Atopic dermatitis (AD) is characterized by reduced barrier function, reduced innate immune activation, and susceptibility to Staphylococcus aureus. Host susceptibility factors are suggested by monogenic disorders associated with AD-like phenotypes and can be medically modulated. S. aureus contributes to AD pathogenesis and can be mitigated by antibiotics and bleach baths. Recent work has revealed that the skin microbiome differs significantly between healthy controls and patients with AD, including decreased Gram-negative bacteria in AD. However, little is known about the potential therapeutic benefit of microbiome modulation. To evaluate whether parameters of AD pathogenesis are altered after exposure to different culturable Gram-negative bacteria (CGN) collected from human skin, CGN were collected from healthy controls and patients with AD. Then, effects on cellular and culture-based models of immune, epithelial, and bacterial function were evaluated. Representative strains were evaluated in the MC903 mouse model of AD. We found that CGN taken from healthy volunteers but not from patients with AD were associated with enhanced barrier function, innate immunity activation, and control of S. aureus. Treatment with CGN from healthy controls improved outcomes in a mouse model of AD. These findings suggest that a live-biotherapeutic approach may hold promise for treatment of patients with AD.

Strain Specific Phage Treatment for Staphylococcus aureus Infection Is Influenced by Host Immunity and Site of Infection.

The response to multi-drug resistant bacterial infections must be a global priority. While mounting resistance threatens to create what the World Health Organization has termed a "post-antibiotic era", the recent discovery that antibiotic use may adversely impact the microbiome adds further urgency to the need for new developmental approaches for anti-pathogen treatments. Methicillin-resistant Staphylococcus aureus (MRSA), in particular, has declared itself a serious threat within the United States and abroad. A potential solution to the problem of antibiotic resistance may not entail looking to the future for completely novel treatments, but instead looking into our history of bacteriophage therapy. This study aimed to test the efficacy, safety, and commercial viability of the use of phages to treat Staphylococcus aureus infections using the commercially available phage SATA-8505. We found that SATA-8505 effectively controls S. aureus growth and reduces bacterial viability both in vitro and in a skin infection mouse model. However, this killing effect was not observed when phage was cultured in the presence of human whole blood. SATA-8505 did not induce inflammatory responses in peripheral blood mononuclear cultures. However, phage did induce IFN gamma production in primary human keratinocyte cultures and induced inflammatory responses in our mouse models, particularly in a mouse model of chronic granulomatous disease. Our findings support the potential efficacy of phage therapy, although regulatory and market factors may limit its wider investigation and use.