ABSTRACT
Misoprostol (MSP) is commonly prescribed in obstetrics and gynecology clinical practice for labor induction, cervical ripening, first-trimester pregnancy termination, and the treatment of postpartum hemorrhage. Furthermore, there is a lack of comprehensive discussion evaluating how different commercially available formulations influence the overall efficacy of MSP, even though reports indicate issues with the quality of these formulations, particularly regarding stability and vaginal absorption processes. This study investigates the stability of MSP under acidic conditions and its in vitro permeation using swine vaginal mucosa. A forced degradation study was conducted using 0.2 M HCl, and a high-efficiency LC method was developed. Three degradation products were identified and characterized using electrospray ionization-high-resolution quadrupole-time-of-flight-MS, with respective m/z values of 391.2508, 405.2705, and 387.2259, respectively. These results suggest that the degradation mechanism involves dehydration of the ß-hydroxy ketone moiety, followed by isomerization to its most resonance-stable form and de-esterification. Finally, the in vitro permeation study revealed that the esterified form of MSP was unable to permeate the mucosa and required prior degradation for any component to be detected in the receptor fluid.
Subject(s)
Drug Stability , Misoprostol , Vagina , Animals , Female , Swine , Vagina/chemistry , Vagina/metabolism , Misoprostol/chemistry , Misoprostol/pharmacokinetics , Misoprostol/analysis , Reproducibility of Results , Chromatography, Liquid/methods , Mass Spectrometry/methods , Mucous Membrane/chemistry , Mucous Membrane/metabolism , Permeability , Liquid Chromatography-Mass SpectrometryABSTRACT
BACKGROUND: There are varying reports of immunohistochemically detected prostatic marker protein distribution in glands associated with the female urethra that may be related to tissue integrity at the time of fixation. AIM: In this study we used tissue derived from rapid autopsies of female patients to determine the distribution of glandular structures expressing prostate-specific antigen (PSA) and prostate-specific acid phosphatase (PSAP) along the female urethra and in surrounding tissues, including the anterior vaginal wall (AVW). METHODS: Tissue blocks from 7 donors that contained the entire urethra and adjacent AVW were analyzed. These tissue samples were fixed within 4-12 hours of death and divided into 5-mm transverse slices that were paraffin embedded. Sections cut from each slice were immunolabeled for PSA or PSAP and a neighboring section was stained with hematoxylin and eosin. The sections were reviewed by light microscopy and analyzed using QuPath software. OBSERVATIONS: In tissue from all donors, glandular structures expressing PSA and/or PSAP were located within the wall of the urethra and were present along its whole length. RESULTS: In the proximal half of the urethra from all donors, small glands expressing PSAP, but not PSA, were observed adjacent to the and emptying into the lumen. In the distal half of the urethra from 5 of the 7 donors, tubuloacinar structures lined by a glandular epithelium expressed both PSA and PSAP. In addition, columnar cells at the surface of structures with a multilayered transitional epithelium in the distal half of the urethra from all donors expressed PSAP. No glands expressing PSA or PSAP were found in tissues surrounding the urethra, including the AVW. CLINICAL IMPLICATIONS: Greater understanding of the distribution of urethral glands expressing prostatic proteins in female patients is important because these glands are reported to contribute to the female sexual response and to urethral pathology, including urethral cysts, diverticula, and adenocarcinoma. STRENGTHS AND LIMITATIONS: Strengths of the present study include the use of rapid autopsy to minimize protein degradation and autolysis, and the preparation of large tissue sections to demonstrate precise anatomical relations within all the tissues surrounding the urethral lumen. Limitations include the sample size and that all donors had advanced malignancy and had undergone previous therapy which may have had unknown tissue effects. CONCLUSION: Proximal and distal glands expressing prostate-specific proteins were observed in tissue from all donors, and these glands were located only within the wall of the urethra.
Subject(s)
Acid Phosphatase , Autopsy , Prostate-Specific Antigen , Urethra , Vagina , Humans , Female , Urethra/pathology , Vagina/pathology , Vagina/chemistry , Prostate-Specific Antigen/analysis , Acid Phosphatase/analysis , Acid Phosphatase/metabolism , Middle Aged , Aged , Protein Tyrosine Phosphatases/metabolism , Protein Tyrosine Phosphatases/analysis , Adult , Biomarkers/metabolism , ImmunohistochemistryABSTRACT
The lower female reproductive tract is notoriously dominated by Lactobacillus species, among which Lactobacillus crispatus emerges for its protective and health-promoting activities. Although previous comparative genome analyses highlighted genetic and phenotypic diversity within the L. crispatus species, most studies have focused on the presence/absence of accessory genes. Here, we investigated the variation at the single nucleotide level within protein-encoding genes shared across a human-derived L. crispatus strain selection, which includes 200 currently available human-derived L. crispatus genomes as well as 41 chromosome sequences of such taxon that have been decoded in the framework of this study. Such data clearly pointed out the presence of intra-species micro-diversities that could have evolutionary significance contributing to phenotypical diversification by affecting protein domains. Specifically, two single nucleotide variations in the type II pullulanase gene sequence led to specific amino acid substitutions, possibly explaining the substantial differences in the growth performances and competition abilities observed in a multi-strain bioreactor culture simulating the vaginal environment. Accordingly, L. crispatus strains display different growth performances, suggesting that the colonisation and stable persistence in the female reproductive tract between the members of this taxon is highly variable.
Subject(s)
Lactobacillus crispatus , Vagina , Lactobacillus crispatus/classification , Lactobacillus crispatus/genetics , Lactobacillus crispatus/growth & development , Lactobacillus crispatus/metabolism , Genome, Bacterial , Evolution, Molecular , Vagina/chemistry , Vagina/microbiology , Humans , Female , Lactobacillus/classification , Lactobacillus/genetics , Carbohydrate MetabolismABSTRACT
Extensive research has explored the causes of embryo losses during early pregnancy by analyzing interaction mechanisms in sows' uterus, ignoring the importance of the lower reproductive tract in pregnancy development regulation. Despite recent progress in understanding the diversity of vaginal microbes under different physiological states, the dynamic of sows' vaginal microbiotas during pregnancy and the interaction between vaginal microbes and the host are poorly understood. Here, we performed a comprehensive analysis of sows' vaginal microbial communities in early pregnancy coupled with overall patterns of vaginal mucosal epithelium gene expression. The vaginal microbiota was analyzed by 16s rRNA or metagenome sequencing, and the vaginal mucosal epithelium transcriptome was analyzed by RNA sequencing, followed by integration of the data layers. We found that the sows' vaginal microbiotas in early pregnancy develop dynamically, and there is a homeostasis balance of Firmicutes and Proteobacteria. Subsequently, we identified two pregnancy-specific communities, which play diverse roles. The microbes in the vagina stimulate the epithelial cells, while vaginal epithelium changes its structure and functions in response to stimulation. These changes produce specific inflammation responses to promote pregnancy development. Our findings demonstrate the interaction between microbes and host in the sow vagina in early pregnancy to promote pregnancy development, meanwhile providing a reference data set for the study of targeted therapies of microbial homeostasis dysregulation in the female reproductive tract. IMPORTANCE This work sheds light on the dynamics of the sow vaginal microbiotas in early pregnancy and its roles in pregnancy development. Furthermore, this study provides insight into the functional mechanisms of reproductive tract microbes by outlining vaginal microbe-host interactions, which might identify new research and intervention targets for improving pregnancy development by modulating lower reproductive tract microbiota.
Subject(s)
Microbiota , Vagina , Pregnancy , Animals , Female , Swine , RNA, Ribosomal, 16S/genetics , Vagina/chemistry , Uterus/chemistry , Microbiota/genetics , MetagenomeABSTRACT
Aerobic bacteria can colonize the female reproductive system with harmful effects, which may lead to miscarriages, premature deliveries or continue of its growing to cause adverse reproductive systems issues. Increasing in the levels of inflammatory markers may be considered a herald of danger. High vaginal swabs were obtained from 85 women. Of these, 67 patients were suffering recurrent vaginitis and symptoms such as itching, irritation, burning, and vaginal discharged, and 18 apparently healthy controls. Swabs were cultured in a suitable media and the cultivated bacteria were diagnosed in the hospital's laboratory. At the same time of collecting the vaginal swabs, 5 ml of venous blood was withdrawn from the patients and controls. An ELISA method was applied to measure the levels of inflammatory cytokines and concentration of vitamin D. The bacterial growth showed six species of isolated bacteria, which were Staphylococcus aureus, Streptococcus spp., E. coli, S. non aureus, Proteus spp. and Klebsiella spp. The first three species were the most prevalent bacteria, and the serum levels of C reactive protein (CRP) and IL-6 were high in female patients infected with these bacteria. CRP was significantly elevated in sera of the patient's group (P= 0.016), while the increase in IL-6 was not significant. Vitamin D was correlated negatively with IL-6 and positively with CRP, but the correlations did not reach statistical significance. In conclusion, rising of CRP could be an expected result to the bacterial colonizing the reproductive system while IL-6 may develop significantly when the aerobic vaginitis continues until triggering one of the infertility issues.
Subject(s)
C-Reactive Protein , Interleukin-6/analysis , Bacteria , C-Reactive Protein/analysis , Escherichia coli , Female , Humans , Interleukin-6/metabolism , Reinfection , Vagina/chemistry , Vagina/microbiology , Vitamin DABSTRACT
Importance: Postmenopausal women with genitourinary symptoms of menopause are often prescribed vaginal estradiol or moisturizer for symptom improvement, but the impact of these treatments on the local microenvironment is poorly understood. Objective: To compare changes in the vaginal microbiota, metabolome, and pH among women using low-dose vaginal estradiol tablet or low pH moisturizer gel for 12-weeks vs low pH placebo. Design, Setting, and Participants: This is a post hoc prespecified secondary analysis of a 12-week multicenter randomized clinical trial among postmenopausal women with moderate to severe genitourinary symptoms. Women were enrolled between April 2016 and February 2017; final follow-up visits occurred in April 2017. Data were analyzed from November 2018 to July 2021. Interventions: Ten-µg vaginal estradiol plus placebo gel vs placebo tablet plus vaginal moisturizer vs dual placebo. Main Outcomes and Measures: The main outcome measures were changes in the diversity and composition of the vaginal microbiota, changes in the metabolome, and pH. Results: Of 302 postmenopausal women from the parent trial, 144 women (mean [SD] age, 61 [4] years) were included in this analysis. After 12 weeks, the microbiota was dominated with Lactobacillus and Bifidobacterium communities among 36 women (80%) in the estradiol group, compared with 16 women (36%) using moisturizer and 13 women (26%) using placebo (P < .001). The composition of vaginal fluid metabolites also varied after 12-weeks among women in the estradiol group with significant changes in 90 of 171 metabolites measured (53%) (P < .001), including an increase in lactate. The 12-week pH among women in the estradiol group was lower vs placebo (median [IQR] pH, 5 [4.5-6.0] vs 6 [5.5-7.0]; P = .005) but not the moisturizer group vs placebo (median [IQR] pH, 6 [5.5-6.5]; P = .28). There was a decrease in pH from baseline to 12-weeks within the moisturizer (median [IQR] pH, 7 [6.0-7.5] vs 6 [5.5-6.5]; P < .001) and placebo (median [IQR] pH, 7 [7.0-7.5] vs 6 [5.5-7.0]; P < .001) groups. Women with high-diversity bacterial communities at baseline exhibited greater median change in pH compared with women with low-diversity communities (median [IQR] change, -1 [-2 to -0.5] vs -0.3 [-1.1 to 0], P = .007). Conclusions and Relevance: This secondary analysis of a randomized clinical trial found that use of vaginal estradiol tablets resulted in substantial changes in the vaginal microbiota and metabolome with a lowering in pH, particularly in women with high-diversity bacterial communities at baseline. Low pH moisturizer or placebo did not significantly impact the vaginal microbiota or metabolome despite lowering the vaginal pH. Estradiol use may offer additional genitourinary health benefits to postmenopausal women. Trial Registration: ClinicalTrials.gov Identifier: NCT02516202.
Subject(s)
Microbiota , Postmenopause , Double-Blind Method , Female , Humans , Metabolome , Middle Aged , Vagina/chemistryABSTRACT
BACKGROUND: 16S ribosomal RNA (rRNA) gene sequence analysis is the common method to identify the bacteria in human vaginal flora. While specific DNA primers were designed to target 16S rRNA gene sequences, DNA fragment analysis using capillary electrophoresis can obtain more accurate qualitative and quantitative information on the microbiome. This study aimed to assess the ability of capillary electrophoresis method to analyze the diversity of vaginal microbiome and provide a theoretical basis for the accurate gene detection of vaginal flora. METHODS: We collected 75 vaginal secretion samples from female outpatients aged 25 - 50, who had undergone rou-tine gynecologic examinations in Fujian provincial hospital from March 2021 to April 2021. Clinical diagnosis was based on the results of microscopic examination of Gram-stained specimens and biochemical tests of bacteria (pH value, catalase, leukocyte esterase, sialidases, ß-glucuronidase, and acetylglucosaminidase). Vaginal secretion samples were collected and then total bacterial DNA was extracted. We used six pairs of fluorescent dye tagged specific primers that were designed based on the 16S rRNA genes of four Lactobacillus species (L. iners, L. crispatus, L. jensenii, L. gasseri), Gardnerella vaginalis, and Atopobium vaginae. PCR products of six species of bacteria were detected and analyzed by a 3130 Genetic Analyzer. RESULTS: Seventy-five samples were divided into two groups according to the vaginal microbiome evaluation, including 50 cases which had vaginal bacteria balance and 25 cases which had vaginal bacteria disorder. PCR amplification of 16S ribosomal RNA genes of L. iners, L. crispatus, L. jensenii, L. gasseri, Gardnerella vaginalis, and Atopobium vaginae was successfully performed on the DNA extracted from vaginal secretion samples. Four Lac-tobacillus species were detected in 4 - 33 cases of "Balance" group, and Gardnerella vaginalis was detected in 23 cases of "Disorder" group and, simultaneously, Atopobium vaginae was detected in 20 cases. CONCLUSIONS: Based on the technique of DNA fragment analysis using capillary electrophoresis method, the most common vaginal bacteria in Chinese healthy women are L. iners and L. crispatus. Gardnerella vaginalis and Atopobium vaginae are the most common pathogenic bacteria detected in the patients who had vaginal bacteria disorder. Using capillary electrophoresis method to detect the vaginal bacteria will be useful for accurate identifica-tion of vaginal microbiome. There will be an application value to find out the composition of the vaginal microbiome rapidly and detect specific gene markers to identify potential pathogenic bacteria when women are at risk of serious illness before they develop obvious symptoms.
Subject(s)
Microbiota , Vaginosis, Bacterial , Adult , Electrophoresis, Capillary , Female , Gardnerella vaginalis/genetics , Humans , Microbiota/genetics , Middle Aged , RNA, Ribosomal, 16S/genetics , Vagina/chemistry , Vagina/microbiology , Vaginosis, Bacterial/diagnosis , Vaginosis, Bacterial/microbiologyABSTRACT
BACKGROUND: Lactobacillus was described as a keystone bacterial taxon in the human vagina over 100 years ago. Using metagenomics, we and others have characterized lactobacilli and other vaginal taxa across health and disease states, including pregnancy. While shifts in community membership have been resolved at the genus/species level, strain dynamics remain poorly characterized. METHODS: We performed a metagenomic analysis of the complex ecology of the vaginal econiche during and after pregnancy in a large U.S. based longitudinal cohort of women who were initially sampled in the third trimester of pregnancy, then validated key findings in a second cohort of women initially sampled in the second trimester of pregnancy. FINDINGS: First, we resolved microbial species and strains, interrogated their co-occurrence patterns, and probed the relationship between keystone species and preterm birth outcomes. Second, to determine the role of human heredity in shaping vaginal microbial ecology in relation to preterm birth, we performed a mtDNA-bacterial species association analysis. Finally, we explored the clinical utility of metagenomics in detection and co-occurrence patterns for the pathobiont Group B Streptococcus (causative bacterium of invasive neonatal sepsis). CONCLUSIONS: Our highly refined resolutions of the vaginal ecology during and post-pregnancy provide insights into not only structural and functional community dynamics, but highlight the capacity of metagenomics to reveal finer aspects of the vaginal microbial ecologic framework. FUNDING: NIH-NINR R01NR014792, NIH-NICHD R01HD091731, NIH National Children's Study Formative Research, Burroughs Wellcome Fund Preterm Birth Initiative, March of Dimes Preterm Birth Research Initiative, NIH-NIGMS (K12GM084897, T32GM007330, T32GM088129).
Subject(s)
Microbiota , Premature Birth , Bacteria , Child , Female , Humans , Infant, Newborn , Lactobacillus/genetics , Microbiota/genetics , Postpartum Period , Pregnancy , Premature Birth/microbiology , RNA, Ribosomal, 16S/genetics , Vagina/chemistryABSTRACT
Condom evidence has become in the past years a very relevant item of evidence in sexual assault or rape cases, being an objective help in the reconstruction of the activity. Traces recovered from a vaginal swab might help to identify whether a condom or other lubricants were used, and thereby possibly confirming or infirming allegations of the parties. However, the interpretation of condom traces can be challenging and requires a detailed understanding of various factors like condom lubricant chemical composition and occurrence on the market, transfer and persistence parameters and background. Herein, we aimed at improving our understanding of factors affecting the transfer variability of condom residues recovered from vaginal matrix. This work employed Diffuse Reflectance Infrared Fourier Transform Spectroscopy (DRIFTS) to provide new data for a characterization of condom lubricants and their traces after a transfer in a vaginal matrix has occurred. Condom traces were recovered from volunteers and the traces characteristics were investigated and analyzed. The effects of donor (condom) and receiver (vaginal matrix) were firstly evaluated, as they are known factors, and these data that could be obtained in real caseworks. Using principal component analysis (PCA), this study highlighted that the effect of the donor was more important than the receiver effect. Vaginal matrix residues were not detected in transferred extracts. The discrimination pattern amongst the donor was found to be indistinguishable from the one obtained on reference material using ATR-FTIR (Attenuated Total Reflectance).
Subject(s)
Condoms , Lubricants/analysis , Vagina/chemistry , Coitus , Female , Forensic Sciences , Humans , Principal Component Analysis , Spectroscopy, Fourier Transform InfraredABSTRACT
Zika virus (ZIKV) is a flavivirus transmitted by mosquitoes of the genus Aedes, but unlike other flaviviruses, ZIKV can be sexually transmitted by vaginal intercourse. The healthy vaginal pH ranges from 4.0 to 6.0, reaching values of 6.0-7.0 after semen deposition. Here, we report that low extracellular pH values (range 6.2-6.6) dramatically increase ZIKV infection on cell lines of different origin including some derived from the female genital tract and monocyte-derived macrophages. Furthermore, low pH significantly increased ZIKV infection of human ectocervix and endocervix cultured ex-vivo. Enhancement of infection by low pH was also observed using different ZIKV strains and distinct methods to evaluate viral infection, i.e. plaque assays, RT-PCR, flow cytometry, and fluorescence microscopy. Analysis of the mechanisms involved revealed that the enhancement of ZIKV infection induced by low pH was associated with increased binding of the viral particles to the heparan sulphate expressed on the target cell surface. Acidosis represents a critical but generally overlooked feature of the female genital tract, with major implications for sexual transmission diseases. Our results suggest that low vaginal pH might promote male-to-female transmission of ZIKV infection.
Subject(s)
Cervix Uteri/chemistry , Vagina/chemistry , Zika Virus Infection/transmission , Zika Virus/pathogenicity , Acidosis , Animals , Cell Line , Cervix Uteri/virology , Chlorocebus aethiops , Female , Heparitin Sulfate/metabolism , Humans , Hydrogen-Ion Concentration , Microscopy, Fluorescence , Vagina/virology , Vero Cells , Zika Virus/geneticsABSTRACT
Female primates signal impending ovulation with a suite of sexual signals. Studies of these signals have focussed on visual, and to a lesser extent, acoustic signals, neglecting olfactory signals. We aimed to investigate the information content of female olfactory signals in captive olive baboons (Papio anubis) and relate these to the female fertile period. We studied eight adult females living in four groups at the CNRS Station de Primatologie, Rousset-sur-Arc, France. We used vaginal cytology to detect ovulation. We investigated the volatile component of odour signals using solid-phase microextraction and gas chromatography-mass spectrometry. We found a total of 74 volatile compounds, of which we tentatively identified 25, including several ketones, alcohols, aldehydes, terpenes, volatile fatty acids and hydrocarbons that have been identified in odour profiles of other primates. Our results show that vaginal odour intensity differs with sexual cycle stage suggesting that odour might play a role in signalling female baboon fertility. We found differences in vaginal odour between females living in all-female and in mixed sex groups but we could not distinguish the effects of group composition, female age and identity. This study of olfactory signalling improves our understanding of how female primates advertise their sexual receptivity.
Subject(s)
Fertility , Odorants/analysis , Olfactory Perception/physiology , Papio anubis/physiology , Sexual Behavior, Animal , Vagina/chemistry , Volatile Organic Compounds/analysis , Animals , Female , OvulationABSTRACT
During pregnancy, the vaginal microbiome plays an important role in both maternal and neonatal health outcomes. Throughout pregnancy, the vaginal microbial composition undergoes significant changes, including a decrease in overall diversity and enrichment with Lactobacillus spp. In turn, the modifications in the microbial profiles are associated with shifts in the composition of vaginal metabolites. In this study, we characterized the vaginal metabolic profiles throughout pregnancy at two different gestational ages, correlating them with a microscopic evaluation of the vaginal bacterial composition. A total of 67 Caucasian pregnant women presenting to the Family Advisory Health Centres of Ravenna (Italy) were enrolled and a vaginal swab was collected at gestational ages 9-13 weeks (first trimester) and 20-24 weeks (second trimester). The composition of the vaginal microbiome was assessed by Nugent score and women were divided in 'H' (normal lactobacilli-dominated microbiota), 'I' (intermediate microbiota), and 'BV' (bacterial vaginosis) groups. Starting from the cell-free supernatants of the vaginal swabs, a metabolomic analysis was performed by means of a 1H-NMR spectroscopy. From the first to the second trimester, a greater number of women showed a normal lactobacilli-dominated microbiota, with a reduction of cases of dysbiosis. These microbial shifts were associated with profound changes in the vaginal metabolic profiles. Over the weeks, a significant reduction in the levels of BV-associated metabolites (e.g. acetate, propionate, tyramine, methylamine, putrescine) was observed. At the same time, the vaginal metabolome was characterized by higher concentrations of lactate and of several amino acids (e.g. tryptophan, threonine, isoleucine, leucine), typically found in healthy vaginal conditions. Over time, the vaginal metabolome became less diverse and more homogeneous: in the second trimester, women with BV showed metabolic profiles more similar to the healthy/intermediate groups, compared to the first trimester. Our data could help unravel the role of vaginal metabolites in the pathophysiology of pregnancy.
Subject(s)
Bacteria/classification , Metabolomics/methods , Pregnancy Trimester, First/metabolism , Pregnancy Trimester, Second/metabolism , Vagina/chemistry , Adult , Amino Acids/analysis , Bacteria/genetics , Bacteria/isolation & purification , DNA, Bacterial/genetics , Female , Humans , Italy , Lactic Acid/analysis , Pregnancy , Proton Magnetic Resonance Spectroscopy , Vagina/microbiology , Young AdultABSTRACT
There are a variety of contraceptive methods that suit individuals' various lifestyles and needs. In the spring of 2020, the U.S. Food and Drug Administration approved Phexxi, a gel-based, nonhormonal, on-demand method of contraception. Phexxi is a pH regulator applied vaginally to create an acidic environment to reduce sperm motility. This article offers an overview of Phexxi, including dosage and administration, potential adverse effects, considerations for special populations, and implications for women's health nurses.
Subject(s)
Contraception , Sperm Motility , Vagina/chemistry , Administration, Intravaginal , Female , Humans , Hydrogen-Ion Concentration , Male , Sperm Motility/drug effects , United StatesABSTRACT
Condom traces are increasingly detected from victims of sexual assault, mostly from vaginal swabs. Protocols have been developed for the analysis of silicone-based condom lubricants using DRIFTS-FTIR and py-GC/MS, but very little research is concerned with the background contribution of the vaginal matrix itself. The present contribution would be an asset for more fundamental research on condom residues in the vaginal matrix, as well as for interpretative purposes in the forensic area. This study investigated vaginal matrix residues using Diffuse Reflectance Fourier Transform Infrared Spectroscopy (DRIFTS-FTIR) and pyrolysis Gas-Chromatography coupled to Mass Spectrometry (py-GC/MS) to obtain fundamental information about the vaginal matrix's initial composition. Differences between women of a given population were investigated as well as the prevalence of silicone-based residues for natural purposes in the population. Apolar fractions of the samples were investigated after extraction with hexane, as it is the one targeted for silicone-based lubricants used in condoms. Infrared spectroscopy outlined the presence of various proteins and lipids in all the samples, and the spectral regions 1000-1850â¯cm-1 and 2700-3600â¯cm-1 were identified as the most relevant zones of the spectra. Pyrolysis-GC results confirmed the presence of lipids, more specifically the presence of cholesterol residues. Chemometrics analyses showed that it was not possible to distinguish the samples based on the qualitative nor semi-quantitative content. This suggest that the same type of compounds are extracted regardless of the donor. None of the samples were found to contain any silicones residues. These results are promising from a forensic evidence interpretation perspective. Further research is required to fully validate such models and assess their robustness and limitation in casework conditions.
Subject(s)
Condoms , Gas Chromatography-Mass Spectrometry , Spectroscopy, Fourier Transform Infrared , Vagina/chemistry , Adolescent , Adult , Female , Forensic Sciences , Humans , Young AdultABSTRACT
Neospora caninum is a protozoan that can cause reproductive problems in several animal species. Although N. caninum infection has been reported in swine, the pathogenesis and clinical signs are not fully known in this species. The objective of this work was to evaluate the effect of experimental infection with tachyzoites of the N. caninum strain Nc1 in swine matrices at different stages of gestation. For that purpose, 12 gilts, seronegative for N. caninum and T. gondii, were selected and allocated into four groups of three animals each. Animals in group A were not inoculated (control) and animals in groups B, C, and D were inoculated intravenously with of 2.9 × 107 tachyzoites, 30 days before conception, and at 45 and 90 days of gestation, respectively. Temperature, heart rate, blood, saliva, and vaginal mucus samples from the animals were collected periodically until the time of delivery for the investigation of IgG and IgM antibodies against N. caninum using IFAT and PCR to detect the parasite DNA. All gilts sero-converted from 5 and 7 DPI (days postinoculation) to IgM and IgG, respectively. Two gilts showed hypothermia on the 5th and 7th DPI, and five inoculated animals had leukocytosis on the 7th DPI. It was possible to detect DNA of N. caninum in samples of saliva (33/84), vaginal mucus (17/84), and blood (2/84). Based on serology (IgM) and PCR, three animals in group B showed evidence of reappearance of the infection during pregnancy. It is concluded that N. caninum can cause clinical signs in infected swine females, in addition to indicating saliva as a suitable diagnostic biological material for the detection of N. caninum DNA in this animal species.
Subject(s)
Coccidiosis/veterinary , Neospora/classification , Pregnancy Complications, Parasitic/veterinary , Swine Diseases/parasitology , Animals , Antibodies, Protozoan/analysis , Antibodies, Protozoan/blood , Coccidiosis/parasitology , DNA, Protozoan/analysis , DNA, Protozoan/blood , Female , Immunoglobulin G/analysis , Immunoglobulin G/blood , Immunoglobulin M/analysis , Immunoglobulin M/blood , Neospora/immunology , Neospora/pathogenicity , Plasma/immunology , Polymerase Chain Reaction/veterinary , Pregnancy , Pregnancy Complications, Parasitic/parasitology , Saliva/immunology , Swine , Vagina/chemistry , Vagina/immunologyABSTRACT
The purpose of the present study was to determine the diagnostic accuracy of vaginal urea and creatinine levels in the detection of premature rupture of membrane (PROM). The Cochrane (central), EMBASE, PubMed, Scopus, and Web of Science were searched for studies published from the inception of the databases up to January 2020. We included published observational full-text articles. The mean differences (MD) and 95% confidence intervals (95% CI) were calculated. The significance level was set as 0.05. Eleven studies (n = 1324) were considered for meta-analysis. Using the bivariate model, the summary estimate of sensitivity and specificity for urea was 0.96 (95% CI: 0.86, 0.98) and 0.93 (95% CI: 0.83, 0.97), respectively. The summary estimate of sensitivity and specificity for creatinine was 0.98 (95% CI: 0.92, 0.99) and 0.97 (95% CI: 0.89, 0.99), respectively. The overall mean of urea and creatinine in the case group was significantly higher than that in the control group (MD = 12.63, 95%, CI [12.01, 13.25]) and (MD = 0.31, 95%, CI [0.29, 0.32]), respectively. The results of this systematic review showed that the mean of urea and creatinine in the case group was significantly higher than that in the control group and the sensitivity and specificity of creatinine is higher than urea in the diagnosis of PROM.
Subject(s)
Body Fluids/chemistry , Creatinine/analysis , Fetal Membranes, Premature Rupture/metabolism , Urea/analysis , Vagina/chemistry , Biomarkers/analysis , Female , Fetal Membranes, Premature Rupture/diagnosis , Humans , Predictive Value of Tests , Pregnancy , Reproducibility of Results , Up-RegulationABSTRACT
Since our previous results suggest that lactoferrin (LF) might have roles in the reproductive process and that its levels might change in the female tract as a response to various factors, the aim of this investigation was to assess whether LF levels in cervical secretions correlate with reproductive parameters from in vitro fertilization (IVF) patients. Cervical fluid samples were obtained from 34 women under 40 years old enrolled for assisted reproduction techniques, and LF concentration was measured. The mean total protein concentration in all cervical fluid samples was 842.8 ± 116.9 µg/mL. The mean concentration of LF was 0.73 ± 0.06 ng LF/µg of total proteins. We observed that higher LF levels in cervical fluid correlated with lower IVF rates when all patients were analyzed; this negative correlation was also sustained when only patients ≥35 years were studied. The mean LF concentration in cervical fluid was significantly lower among patients with normal IVF rates than in those with values 50% or less. Using a LF cutoff value of 0.83 ng/µg of total proteins, the study revealed a significant association between the LF levels below 0.83 ng/µg of total proteins and IVF rates above 50%. LF levels in cervical mucus could potentially be used as a marker of fertilization outcome.
Subject(s)
Body Fluids/chemistry , Cervix Mucus/chemistry , Fertilization in Vitro , Lactoferrin/analysis , Vagina/chemistry , Adult , Enzyme-Linked Immunosorbent Assay , Female , HumansABSTRACT
BACKGROUND: Microbiome and metagenomic studies have given rise to a new understanding of microbial colonization of various human tissues and their ability to impact our health. One human microbiome growing in notoriety, the vaginal microbiome, stands out given its importance for women's health, and is peculiar in terms of its relative bacterial composition, including its simplicity and typical domination by a small number of Lactobacillus species. The loss of Lactobacillus dominance is associated with disorders such as bacterial vaginosis, and efforts are now underway to understand the ability of Lactobacillus species to colonize the vaginal tract and adapt to this dynamic and acidic environment. Here, we investigate how various Lactobacillus species often isolated from the vaginal and intestinal cavities genomically and transcriptionally respond to iterative growth in simulated vaginal fluid. RESULTS: We determined the genomes and transcriptomes of L. acidophilus, L. crispatus, L. fermentum, L. gasseri, and L. jensenii and compared profiles after 50, 100, 500, and 1000 generations of iterative passages in synthetic vaginal fluid. In general, we identified relatively few genetic changes consisting of single nucleotide polymorphisms, with higher counts occurring more frequently in non-vaginal isolated species. Transcriptional profiles were more impacted over time and tended to be more extensive for species that typically do not dominate the vaginal tract, reflecting a more extensive need to adapt to a less familiar environment. CONCLUSIONS: This study provides insights into how vaginal and non-vaginal Lactobacillus species respond and adapt to a simulated vaginal environment. Overall, trends indicate high genomic stability for all species involved, with more variability in the transcriptome especially for non-dominant species of the vaginal tract.
Subject(s)
Adaptation, Physiological , Body Fluids/microbiology , Lactobacillus/physiology , Vagina/microbiology , Adaptation, Physiological/genetics , Body Fluids/chemistry , Female , Genome, Bacterial , Genomic Instability , Humans , Lactobacillus/classification , Lactobacillus/genetics , Lactobacillus/growth & development , Species Specificity , Transcriptome , Vagina/chemistryABSTRACT
INTRODUCTION: The majority of new HIV infections occur through mucosal transmission. The availability of readily applicable and accessible platforms for anti-retroviral (ARV) delivery is critical for the prevention of HIV acquisition through sexual transmission in both women and men. There is a compelling need for developing new topical delivery systems that have advantages over the pills, gels and rings, which currently fail to guarantee protection against mucosal viral transmission in vulnerable populations due to lack of user compliance. The silk fibroin (SF) platform offers another option that may be better suited to individual circumstances and preferences to increase efficacy through user compliance. The objective of this study was to test safety and efficacy of SF for anti-HIV drug delivery to mucosal sites and for viral prevention. METHODS: We formulated a potent HIV inhibitor Griffithsin (Grft) in a mucoadhesive silk fibroin (SF) drug delivery platform and tested the application in a non-human primate model in vivo and a pre-clinical human cervical and colorectal tissue explant model. Both vaginal and rectal compartments were assessed in rhesus macaques (Mucaca mulatta) that received SF (n = 4), no SF (n = 7) and SF-Grft (n = 11). In this study, we evaluated the composition of local microbiota, inflammatory cytokine production, histopathological changes in the vaginal and rectal compartments and mucosal protection after ex vivo SHIV challenge. RESULTS: Effective Grft release and retention in mucosal tissues from the SF-Grft platform resulted in protection against HIV in human cervical and colorectal tissue as well as against SHIV challenge in both rhesus macaque vaginal and rectal tissues. Mucoadhesion of SF-Grft inserts did not cause any inflammatory responses or changes in local microbiota. CONCLUSIONS: We demonstrated that in vivo delivery of SF-Grft in rhesus macaques fully protects against SHIV challenge ex vivo after two hours of application and is safe to use in both the vaginal and rectal compartments. Our study provides support for the development of silk fibroin as a highly promising, user-friendly HIV prevention modality to address the global disparity in HIV infection.
Subject(s)
Anti-HIV Agents/administration & dosage , Fibroins , HIV Infections/prevention & control , Lectins/administration & dosage , Plant Lectins/administration & dosage , Simian Acquired Immunodeficiency Syndrome/prevention & control , Animals , Anti-HIV Agents/analysis , Anti-HIV Agents/pharmacokinetics , Biocompatible Materials , Cervix Uteri/virology , Colon/virology , Female , Gastrointestinal Microbiome/drug effects , HIV/drug effects , Humans , Lectins/analysis , Lectins/pharmacokinetics , Macaca mulatta , Microbiota/drug effects , Mucous Membrane/chemistry , Pharmaceutical Vehicles , Plant Lectins/analysis , Plant Lectins/pharmacokinetics , Rectum/chemistry , Rectum/microbiology , Rectum/virology , Vagina/chemistry , Vagina/microbiologyABSTRACT
The most important proteins regulating cellular zinc homeostasis belong to two protein families of zinc transporters, the solute carrier family 30 (SLC30A) and solute carrier family 39 (SLC39A). We aimed to identify single nucleotide polymorphisms (SNPs) of the SLC30A and SLC39A genes and its association with blood and vaginal tissue zinc levels since vaginal tissue zinc level may play a role in vaginal remodeling and pathological conditions of the vagina. Blood and vaginal tissue samples were collected from women undergoing surgery for benign gynecological reasons. SNPs of twenty-four zinc transporters were determined by PCR/Sequence method, and the concentration of zinc was assessed by inductively coupled plasma optical emission spectrometry. Sequencing of selected exons revealed 16 SNPs, including five previously unidentified SNPs. Our data showed an association between the number of SNPs (more than six SNPs vs. less than six) per patient and high zinc vaginal tissue levels (67% vs. 33%, p < 0.01). The SLC39A4 SNP 590c A (rs17855765) was significantly more frequent in the group of women with high zinc vaginal tissue levels compared to the group without SNP (93% vs. 7%, p = 0.02). Also, our analysis revealed that the number of SNPs in SLC39A4 was significantly more frequent in patients with low zinc blood levels (76% vs. 24%, p = 0.01). Our findings indicate that different SNPs of the zinc transporter genes may have a significant effect on the blood and vaginal tissue zinc levels.