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Microbiome May 2020Gender reassignment surgery is a procedure some transgender women (TW) undergo for gender-affirming purposes. This often includes the construction of a neovagina using...
BACKGROUND
Gender reassignment surgery is a procedure some transgender women (TW) undergo for gender-affirming purposes. This often includes the construction of a neovagina using existing penile and scrotal tissue and/or a sigmoid colon graft. There are limited data regarding the composition and function of the neovaginal microbiome representing a major gap in knowledge in neovaginal health.
RESULTS
Metaproteomics was performed on secretions collected from the neovaginas (n = 5) and rectums (n = 7) of TW surgically reassigned via penile inversion/scrotal graft with (n = 1) or without (n = 4) a sigmoid colon graft extension and compared with secretions from cis vaginas (n = 32). We identified 541 unique bacterial proteins from 38 taxa. The most abundant taxa in the neovaginas were Porphyromonas (30.2%), Peptostreptococcus (9.2%), Prevotella (9.0%), Mobiluncus (8.0%), and Jonquetella (7.2%), while cis vaginas were primarily Lactobacillus and Gardnerella. Rectal samples were mainly composed of Prevotella and Roseburia. Neovaginas (median Shannon's H index = 1.33) had higher alpha diversity compared to cis vaginas (Shannon's H = 0.35) (p = 7.2E-3, Mann-Whitney U test) and were more similar to the non-Lactobacillus dominant/polymicrobial cis vaginas based on beta diversity (perMANOVA, p = 0.001, r = 0.342). In comparison to cis vaginas, toll-like receptor response, amino acid, and short-chain fatty acid metabolic pathways were increased (p < 0.01), while keratinization and cornification proteins were decreased (p < 0.001) in the neovaginal proteome.
CONCLUSIONS
Penile skin-lined neovaginas have diverse, polymicrobial communities that show similarities in composition to uncircumcised penises and host responses to cis vaginas with bacterial vaginosis (BV) including increased immune activation pathways and decreased epithelial barrier function. Developing a better understanding of microbiome-associated inflammation in the neovaginal environment will be important for improving our knowledge of neovaginal health. Video Abstract.
Topics: Adult; Bacteria; Female; Humans; Male; Microbiota; Middle Aged; Sex Reassignment Surgery; Transgender Persons; Vagina
PubMed: 32370783
DOI: 10.1186/s40168-020-00804-1 -
The Journal of Investigative Dermatology Sep 2020Hidradenitis suppurativa (HS) is a chronic inflammatory disease of the skin associated with specific lesional dysbiotic features. We studied the microbiome of clinically... (Observational Study)
Observational Study
The Surface Microbiome of Clinically Unaffected Skinfolds in Hidradenitis Suppurativa: A Cross-Sectional Culture-Based and 16S rRNA Gene Amplicon Sequencing Study in 60 Patients.
Hidradenitis suppurativa (HS) is a chronic inflammatory disease of the skin associated with specific lesional dysbiotic features. We studied the microbiome of clinically unaffected typical HS sites (armpits, inguinal folds, and gluteal clefts) in 60 patients with HS and 17 healthy controls. A total of 192 samples obtained by swabbing were analyzed by bacterial cultures. Of these, 116 randomly selected samples were studied by 16S rRNA gene amplicon sequencing. Patients and controls showed similar characteristics, except for smoking (87% vs. 6%, respectively). HS skinfolds were characterized by an increased abundance of anaerobes, predominantly Prevotella, but also Actinomyces, Campylobacter ureolyticus, and Mobiluncus, contrasting with a lower abundance of skin commensals such as Staphylococcus epidermidis, a major component of the skin microbiome; Kocuria; and Micrococcus luteus. The following three independent factors were associated with an abundance of high anaerobes by multivariate analysis: samples originating from patients with HS patients (P = 2.1 × 10); body mass index (P = 5 × 10); and the sampling site, the gluteal cleft being the most anaerobic area, followed by inguinal folds and axilla (P = 3 × 10). The microbiome of clinically unaffected HS skinfolds is reminiscent, albeit to a minor extent, of the microbiome of chronic suppurative HS lesions and may fuel inflammation at a preclinical stage of the disease.
Topics: Adult; Bacteria, Anaerobic; Cross-Sectional Studies; DNA, Bacterial; Female; Healthy Volunteers; Hidradenitis Suppurativa; Humans; Male; Microbiota; Middle Aged; Prospective Studies; RNA, Ribosomal, 16S; Sequence Analysis, DNA; Severity of Illness Index; Skin; Young Adult
PubMed: 32339539
DOI: 10.1016/j.jid.2020.02.046 -
Anaerobe Feb 2020Recent human clinical studies have identified Mobiluncus mulieris, a fastidious strict anaerobic bacterium present in the cervicovaginal (CV) space, as being strongly...
Microbial supernatants from Mobiluncus mulieris, a bacteria strongly associated with spontaneous preterm birth, disrupts the cervical epithelial barrier through inflammatory and miRNA mediated mechanisms.
Recent human clinical studies have identified Mobiluncus mulieris, a fastidious strict anaerobic bacterium present in the cervicovaginal (CV) space, as being strongly associated with spontaneous preterm birth (sPTB). However, the molecular mechanisms that underlie this association remain unknown. As disruption of the cervical epithelial barrier has been shown to contribute to the premature cervical remodeling that precedes sPTB, we hypothesize that M. mulieris, a microbe strongly associated with sPTB in humans, has the ability to alter cervical epithelial function. We investigated if bacteria-free supernatants of M. mulieris were able to disrupt the cervical epithelial barrier through immunological and epigenetic based mechanisms in an in vitro model system. Ectocervical cells were treated with supernatant from cultured M. mulieris and epithelial cell permeability, immune cytokines and microRNAs (miRNAs) were investigated. M. mulieris supernatant significantly increased cell permeability and the expression of two inflammatory mediators associated with cervical epithelial breakdown, IL-6 and IL-8. Moreover, treatment of the ectocervical cells with the M. mulieris supernatant also increased the expression of miRNAs that have been associated with either sPTB or a shorter gestational length in humans. Collectively, these results suggest that M. mulieris induces molecular and functional changes in the cervical epithelial barrier thought to contribute to the pathogenesis of sPTB, which allows us to hypothesize that targeting CV bacteria such as M. mulieris could provide a therapeutic opportunity to reduce sPTB rates.
Topics: Actinomycetales Infections; Biomarkers; Cell Membrane Permeability; Culture Media, Conditioned; Cytokines; Epithelial Cells; Female; Gene Expression Regulation; Humans; Inflammation Mediators; MicroRNAs; Mobiluncus; Mucous Membrane; Premature Birth
PubMed: 31760081
DOI: 10.1016/j.anaerobe.2019.102127 -
Immunological Investigations Aug 2020We aimed to study the relation between both bacterial colonization of the uterine endometrium & endometrial miR-17-5p levels and endometriosis, and then to evaluate...
UNLABELLED
We aimed to study the relation between both bacterial colonization of the uterine endometrium & endometrial miR-17-5p levels and endometriosis, and then to evaluate endometrial miR-17-5p as a biomarker of endometriosis. A comparative observational study was carried over 51 endometriosis patients and 51 controls admitted into Obstetrics and Gynecology department, Mansoura Faculty of Medicine. Endometrial tissue samples were collected and aimed for bacterial culture and identification of resulting organisms besides estimation of tissue levels of microRNA-17-5p by quantitative real time PCR. G. vaginalis, S. agalactiae, S. aureus, Mobiluncus and E. coli were associated with endometriosis. MicroRNA-17-5p was up-regulated in endometriosis patients ( value was <0.0001*). Its sensitivity and specificity were 90% and 76.5%. MiR-17-5p showed higher results in culture positive than negative cases. On studying the relation between the positivity of endometrial tissue culture and miR-17-5p and so endometriosis, value was <0.0001*. We concluded that G. vaginalis, S. agalactiae, S. aureus, Mobiluncus and E. coli were associated with development of endometriosis. Endometrial miR-17-5p was elevated in association to positive detection of bacterial species. MiR-17-5p might be a bio- marker of endometriosis.
ABBREVIATIONS
CFU/ml: Colony Forming Unit per Milliliter; miR-17-5p: MicroRNA-17-5p; qRT PCR: Quantitative Real Time Polymerase Chain Reaction.
Topics: Adult; Bacterial Infections; Disease Susceptibility; Egypt; Endometriosis; Endometrium; Female; Humans; MicroRNAs; Middle Aged; Population Surveillance; ROC Curve; Risk Assessment; Risk Factors; Young Adult
PubMed: 31747809
DOI: 10.1080/08820139.2019.1693592 -
Structure determination of CAMP factor of Mobiluncus curtisii and insights into structural dynamics.International Journal of Biological... May 2020Bacterial vaginosis (BV) is a common type of vaginal inflammation caused by a proliferation of pathogenic bacteria, among which Mobiluncus curtisii. In our previous...
Bacterial vaginosis (BV) is a common type of vaginal inflammation caused by a proliferation of pathogenic bacteria, among which Mobiluncus curtisii. In our previous studies on M. curtisii genome, we identified the presence of a genomic fragment encoding a 25 kDa pore-forming toxin, the CAMP factor, which is known to be involved in the synergistic lysis of erythrocytes namely CAMP reaction. However, whether this hypothetical gene product has hemolytic activity is unknown. Moreover, its relative structure and function are not yet solved. Here we found that the M. curtisii CAMP factor is a monomer at pH 4.4 and oligomer at pH > 4.6. Hemolysis assays showed that M. curtisii CAMP factor could lyse sheep red blood cells efficiently in pH 5.4-7.4. Negative staining electron microscope analysis of the CAMP factor revealed ring-like structures at pH above 4.6. Additionally, the crystal structure of M. curtisii CAMP factor, determineded at 1.85 Å resolution, reveals a 5 + 3 helix motif. Further functional analysis suggested that the structural rearrangement of the N-terminal domain might be required for protein function. In conclusion, this structure-function relationship study of CAMP factor provides a new perspective of the M. curtisii role in BV development.
Topics: Actinomycetales Infections; Animals; Bacterial Proteins; Erythrocytes; Female; Humans; Mobiluncus; Molecular Dynamics Simulation; Pore Forming Cytotoxic Proteins; Protein Domains; Sheep; Structure-Activity Relationship; Vaginosis, Bacterial
PubMed: 31739050
DOI: 10.1016/j.ijbiomac.2019.10.107 -
Nature Communications Mar 2019Failure to predict and understand the causes of preterm birth, the leading cause of neonatal morbidity and mortality, have limited effective interventions and...
Failure to predict and understand the causes of preterm birth, the leading cause of neonatal morbidity and mortality, have limited effective interventions and therapeutics. From a cohort of 2000 pregnant women, we performed a nested case control study on 107 well-phenotyped cases of spontaneous preterm birth (sPTB) and 432 women delivering at term. Using innovative Bayesian modeling of cervicovaginal microbiota, seven bacterial taxa were significantly associated with increased risk of sPTB, with a stronger effect in African American women. However, higher vaginal levels of β-defensin-2 lowered the risk of sPTB associated with cervicovaginal microbiota in an ethnicity-dependent manner. Surprisingly, even in Lactobacillus spp. dominated cervicovaginal microbiota, low β-defensin-2 was associated with increased risk of sPTB. These findings hold promise for diagnostics to accurately identify women at risk for sPTB early in pregnancy. Therapeutic strategies could include immune modulators and microbiome-based therapeutics to reduce this significant health burden.
Topics: Adult; Bayes Theorem; Biomarkers; Black People; Case-Control Studies; Cervix Uteri; Female; Gene Expression; Humans; Immunity, Innate; Infant, Newborn; Lactobacillus; Microbiota; Mobiluncus; Pregnancy; Premature Birth; Prognosis; Risk; Vagina; White People; beta-Defensins
PubMed: 30899005
DOI: 10.1038/s41467-019-09285-9 -
Oncology Nursing Forum Mar 2019To characterize the vaginal microbiome using QIIME 2™ (Quantitative Insights Into Microbial Ecology 2) in women with gynecologic cancer. (Comparative Study)
Comparative Study
OBJECTIVES
To characterize the vaginal microbiome using QIIME 2™ (Quantitative Insights Into Microbial Ecology 2) in women with gynecologic cancer.
SAMPLE & SETTING
19 women with gynecologic cancer before and after radiation therapy at a comprehensive cancer center in Atlanta, Georgia.
METHODS & VARIABLES
This pilot study analyzed vaginal microbiome communities using a microbiome analysis pipeline, beginning with 16S rRNA gene sequencing and processing through use of a bioinformatics pipeline to downstream microbial statistical analysis.
RESULTS
The findings showed the methods to be robust, and most women with gynecologic cancer showed depletion of Lactobacillus. Compared to those pre-radiation therapy, women post-radiation therapy showed higher abundances of Mobiluncus, Atopobium, and Prevotella but lower abundances of Lactobacillus, Gardnerella, and Peptostreptococcus, which are associated with bacterial vaginosis.
IMPLICATIONS FOR NURSING
This study presents the fundamentals of human microbiome data collection and analysis methods to inform nursing science. Assessing the vaginal microbiome provides a potential pathway to develop interventions to ameliorate dysbiosis of the vaginal microbiome.
Topics: Adult; Aged; Female; Genital Neoplasms, Female; Georgia; Humans; Microbiota; Middle Aged; Pilot Projects; RNA, Ribosomal, 16S; Vagina
PubMed: 30767956
DOI: 10.1188/19.ONF.E48-E59 -
Annals of Clinical Microbiology and... Nov 2018The long-term use of intrauterine devices (IUDs) may lead to biofilm formation on the surface. The aim of this study was to perform the culture- and PCR-based detection...
Culture- and PCR-based detection of BV associated microbiological profile of the removed IUDs and correlation with the time period of IUD in place and the presence of the symptoms of genital tract infection.
OBJECTIVES
The long-term use of intrauterine devices (IUDs) may lead to biofilm formation on the surface. The aim of this study was to perform the culture- and PCR-based detection of bacteria/fungi from the biofilm of the removed IUDs with different time periods in place.
METHODS
For a 2-year period, 100 IUD users were involved in the study. In the majority of the cases, IUDs were removed because of the patients' complaints. Beside the aerobic and anaerobic culture, species-specific PCR was carried out to detect Chlamydia trachomatis Neisseria gonorrhoeae and the "signalling" bacteria of bacterial vaginosis (BV) in the biofilm removed by vortexing.
RESULTS
Sixty-eight percent of IUDs were used for more than 5 years, 32% were removed after 10 years in place. In 28% of the IUDs ≥ 3 different anaerobic species typically found in BV with or without other aerobic bacteria were found by culture method. Streptococcus agalactiae (14%) and Actinomyces spp. (18%) were also isolated frequently. The PCR detection of Gardnerella vaginalis, Atopobium vaginae, Mobiluncus spp. and Ureaplasma urealyticum were 62%, 32%, 23% and 16%, respectively. Seventy-six percent of the IUDs were PCR positive at least for one "signalling" bacterium of BV. C. trachomatis was detected by PCR only in one IUD together with other aerobic and anaerobic bacteria, while the presence of N. gonorrhoeae could not be confirmed from the biofilm of these removed devices.
CONCLUSION
Sexually transmitted infections (STI)-related bacteria-except for one patient-were not detected on the IUDs removed due to different reasons including clinical symptoms of infection. Presence of any BV "signaling" anaerobic bacteria were detected in a much higher number in the biofilm of the removed IUDs by PCR-based method compared to use culture method (76 versus 28 samples). Different aerobic and anaerobic bacteria colonized an equal number of IUDs, independent of the time-period in place, which may be relevant, if the IUD is removed due to planned pregnancy or due to a fear from upper genital tract infection caused by anaerobic bacteria including Actinomyces spp.
Topics: Adult; Aged; Biofilms; Female; Humans; Intrauterine Devices; Middle Aged; Reproductive Tract Infections; Time Factors; Vaginosis, Bacterial; Young Adult
PubMed: 30466466
DOI: 10.1186/s12941-018-0293-6 -
European Journal of Obstetrics,... Sep 2018The bladder is not sterile but contains a healthy community of microbes termed the microbiome. Alterations in the bladder microbiome have been demonstrated in disease...
OBJECTIVES
The bladder is not sterile but contains a healthy community of microbes termed the microbiome. Alterations in the bladder microbiome have been demonstrated in disease states such as the overactive bladder. The microbiome in other anatomical niches is known to alter with age eg the vagina. The objective of this study was to identify if the bladder microbiome in healthy women varies with age and menopausal status.
STUDY DESIGN
Urine from 79 healthy women attending secondary care gynaecological clinics with no urinary symptoms provided clean catch mid-stream urine specimens. Urine was centrifuged and the resultant pellet was re-suspended and inoculated onto chocolate agar plates and cultured under either aerobic or anaerobic conditions. Morphologically different colonies were purity plated and 16 s rRNA gene sequencing was performed. A microbe genomic basic local alignment search tool (BLAST) was used to identify the genus of the bacteria.
RESULTS
There was no significant correlation between the age of a woman and the number of different genera identified (r=-0.034, p = 0.79). There were few significant differences in the frequency with which the majority of organisms were found in pre and post-menopausal women. The exceptions however were lactobacillus, which was more common in pre-menopausal women (31 vs 3 p = 0.002) and Mobiluncus, which was more common in post-menopausal women (0 vs 3 p = 0.02).
CONCLUSIONS
There was no significant correlation between patient age and diversity of the bladder microbiome but large numbers of different organisms were identified. Significant differences were however observed for Lactobacillus which is more common in pre-menopausal women and Mobiluncus which is more common in post-menopausal women.
Topics: Adolescent; Adult; Aged; Aged, 80 and over; Aging; Female; Humans; Menopause; Microbiota; Middle Aged; Urinary Bladder; Urine; Young Adult
PubMed: 29936400
DOI: 10.1016/j.ejogrb.2018.06.011 -
PeerJ 2018Bacterial vaginosis (BV) is a microbial imbalance (i.e., dysbiosis) that can produce serious medical effects in women at childbearing age. Little is known, however,...
BACKGROUND
Bacterial vaginosis (BV) is a microbial imbalance (i.e., dysbiosis) that can produce serious medical effects in women at childbearing age. Little is known, however, about the incidence of BV or vaginal microbiota dysbiosis in pregnant teenagers in low and middle-income countries such as Ecuador. The scope of this exploratory analysis was to study the relationship between epidemiologic and microbial risk factors. Among the microbiology risk factors this study investigated five species, two of them know in preview studies as microbiology risk factors for BV development ( and ), and the last three known for being associated with a healthy vaginal tract (, and ). In addition, fastidious anaerobes known to be microbial risk factors for BV development in pregnant teenagers were searched as well, more exactly, , and .
METHODS
Ninety-five healthy adolescent pregnant women, visiting a secondary level hospital in Quito, Ecuador, were enrolled into the study in 2015. The enrolled patients were between 10 to 13 weeks of pregnancy. Four epidemiological risk factors were collected in a survey: age, civil status, sexual partners and condom use. Also, vaginal pH was measured as a health risk factor. DNA was extracted from endocervical and exocervical epithelia from all the patients' samples. PCR analysis was performed in order to characterize the presence of the eight bacterial species known as risk factors for BV development, targeting three anaerobes and five species. Univariate and multivariate analysis were performed to identify associated factors for the presence of anaerobic species using logistic regression.
RESULTS
The 95 vaginal microflora samples of these teenagers were analyzed. Two of the bacterial species known to cause BV: (100%) and (93.7%) were found in high prevalence. Moreover, the most predominant bacterial species found in the pregnant teenagers' vaginal tract were (92.6%), (89.5%) and (87.4%). In addition, the average vaginal pH measured in the study population was 5.2, and high pH was associated with the presence of the three-anaerobic species ( = 0.001). Finally, 's presence in the study decreased in 72% the occupation of the three anaerobes.
DISCUSSION
This work identified a high pH as a risk factor for BV anaerobes' presence in adolescent pregnant women. Moreover, this study identified and to be the most abundant species in our study population. From all fastidious anaerobes analyzed in this study, was present in all pregnant teenagers. To conclude, could be a potential healthy vaginal microbiota candidate in pregnant teenagers and should be further analyzed in future studies.
PubMed: 29492333
DOI: 10.7717/peerj.4317