Cervical Fluids Research Articles

Woman’s microbiome and obstetrical and perinatal risks: what do they have in common?

The article is devoted to the review of modern scientific publications about the microbiome of a woman and its importance in obstetric and perinatal practice. The composition of the microbiome is changed both in the process of human evolution and personally and in different periods of a person’s life. However, on the other hand, the microbiome is influenced by the specific environment in which a person lives. It consists of a certain collection of genomes of all microorganisms. Complications during pregnancy occur frequently, according to statistics – in every sixth pregnant woman, and pose a danger to the health of the mother and child.During pregnancy, the richness and diversity of the vaginal microbiome decreases, and different species of Lactobacillus play a dominant role. The microbiome stability is hypothesized to be associated with higher levels of estrogen concentration, absence of menstruation, and modification of cervical and vaginal fluids. During an uncomplicated pregnancy, the taxonomic composition of the vaginal microbiota remains stable with the exception of the period before labor, when there is an increase in microbial diversity. This makes the vaginal microbiome similar to that of a non-pregnant woman and is believed to be a trigger for the labor process to begin.The recent studies have determined a relationship between changes in the microbiome during pregnancy and the occurrence of complications in these women. Any change or disturbance in the microbiome balance can be involved in inflammatory processes, which can potentially have adverse effects on the course of pregnancy. A major interaction between the oral microbiome and preeclampsia involves the production of nitrogen derivatives, especially nitric oxide (NO). NO is produced from L-arginine via NO synthases (NOS), and is involved in vascular processes, especially vasodilation and tissue protection. Also, NO is indirectly involved in the processes that occur in the presence of chronic cardiovascular diseases, including hypertension and preeclampsia.In recent years, scientists have begun actively to study a correlation between changes in the gut microbiome and various nosologies regarding gestational diabetes. There is not much of this data at the moment. The gut microbiome is known to modulate insulin resistance and the inflammatory response, and changes in the microbiome may be associated with metabolic diseases. It is hypothesized that changes in the gut microbiome may lead to metabolic diseases through several mechanisms: abnormal intestinal permeability, increased absorption of lipopolysaccharide, abnormal production of short-chain fatty acids, altered conversion of primary bile acids, and enhanced production of bacterial toxins (e.g., trimethylamine-N-oxide).During full-term pregnancy, the “dynamic stability” of the vaginal microbiome is described as a higher concentration of Lactobacillus species starting at 20 weeks of gestation due to increased glycogen availability and a less complex and diverse microflora. This condition may be due to the lack of cyclical hormonal changes during pregnancy. Lactobacillus and small bacterial diversity are thought to be critical factors for the initiation of labor at the end of the pregnancy.Future therapeutic strategies are likely to be applied to modulate the composition of the microbiome, including the use of pro- and prebiotics and dietary modification. However, further research is needed to identify specific tools that can be used to develop personalized therapeutic approaches for individualized prevention and treatment.

Cervical Fluids Are a Source of Protein Biomarkers for Early, Non-Invasive Endometrial Cancer Diagnosis.

Abnormal uterine bleeding is the main symptom of endometrial cancer (EC), but it is highly nonspecific. This represents a huge burden for women's health since all women presenting with bleeding will undergo sequential invasive tests, which are avoidable for 90-95% of those women who do not have EC. This study aimed to evaluate the potential of cervical samples collected with five different devices as a source of protein biomarkers to diagnose EC. We evaluated the protein quantity and the proteome composition of five cervical sampling methods. Samples collected with a Rovers Cervex Brush® and the HC2 DNA collection device, Digene, were the most suitable samples for EC proteomic studies. Most proteins found in uterine fluids were also detected in both cervical samples. We then conducted a clinical retrospective study to assess the expression of 52 EC-related proteins in 41 patients (22 EC; 19 non-EC), using targeted proteomics. We identified SERPINH1, VIM, TAGLN, PPIA, CSE1L, and CTNNB1 as potential protein biomarkers to discriminate between EC and symptomatic non-EC women with abnormal uterine bleeding in cervical fluids (AUC > 0.8). This study opens an avenue for developing non-invasive protein-based EC diagnostic tests, which will improve the standard of care for gynecological patients.

Congenital Cytomegalovirus

Congenital Cytomegalovirus

Macrophage inflammatory protein-1α in amniotic and cervical fluids in spontaneous preterm labor with intact membranes with respect to intra-amniotic inflammation

Objective Macrophage inflammatory protein 1α is a chemokine produced by various immune, epithelial, mesothelial, and fibroblast cells after exposure to bacterial lipopolysaccharide or pro-inflammatory molecules. The primary aim of this study was to determine MIP-1α concentrations in amniotic and cervical fluids from pregnancy with spontaneous preterm labor with intact membranes (PTL) with respect to the presence of intra-amniotic infection (both microbial invasion of the amniotic cavity and intra-amniotic inflammation) and sterile intra-amniotic inflammation (intra-amniotic inflammation alone). The secondary aim was to assess the diagnostic indices of MIP-1α in predicting intra-amniotic infection. Materials and methods Seventy-four women with PTL were included in this study. Paired amniotic and cervical fluid samples were obtained using transabdominal amniocentesis and a Dacron polyester swab, respectively. Microbial invasion of the amniotic cavity was diagnosed based on a combination of culture and molecular biology methods. The concentration of IL-6 in the amniotic and cervical fluids was measured using an automated electrochemiluminescence immunoassay method. Intra-amniotic inflammation was defined as an amniotic fluid IL-6 concentration of ≥3000 pg/mL. The MIP-1α concentrations in the samples were assessed using an enzyme-linked immunosorbent assay. Results A difference in amniotic fluid MIP-1α was observed among women with intra-amniotic infection, sterile intra-amniotic inflammation, and negative amniotic fluid (infection: median 1779.0 pg/mL; sterile, median 102.7 pg/mL; negative, median 19.9 pg/mL; p < .0001). No difference in the concentrations of MIP-1α was identified in cervical fluid after adjustment for gestational age at sampling (infection: median 77.7 pg/mL, sterile: median 152.7 pg/mL, negative: median 18.0 pg/mL; p = .30). The presence of intra-amniotic infection was associated with elevated MIP-1α concentrations in amniotic fluid (presence: 1779.0 pg/mL vs. absence: 26.3 pg/mL, p < .0001, area under receiver operating characteristic curve = 0.87). Conclusions In PTL pregnancies with the presence of intra-amniotic infection, the concentration of MIP-1α is elevated in amniotic fluid but not in cervical fluid. Amniotic fluid MIP-1α may provide a useful marker for intra-amniotic infection in women with PTL.

IgGFc-binding protein in pregnancies complicated by spontaneous preterm delivery: a retrospective cohort study

To determine the IgGFc-binding protein (FcgammaBP) concentration in amniotic and cervical fluids in preterm prelabor rupture of membranes (PPROM) and preterm labor with intact membranes (PTL) and to assess the diagnostic indices of FcgammaBP to predict intra-amniotic infection (the presence of both microbial invasion of the amniotic cavity and intra-amniotic inflammation). In this study, we included 170 and 79 women with PPROM and PTL, respectively. Paired cervical and amniotic fluid samples were obtained using a Dacron polyester swab and transabdominal amniocentesis, respectively. The FcgammaBP concentrations in the samples were assessed using an enzyme-linked immunosorbent assay. The presence of intra-amniotic infection was associated with elevated FcgammaBP concentrations in pregnancies with PPROM and PTL [PPROM—presence: 86 ng/mL vs. absence: 13 ng/mL, p < 0.0001, area under receiver operating characteristic curve (AUC) = 0.94; PTL—presence: 140 ng/mL vs. absence: 22 ng/mL, p < 0.0001, AUC = 0.86]. In cervical fluid, the concentrations of FcgammaBP were elevated in the presence of intra-amniotic infection in pregnancies with PPROM only (presence: 345 ng/mL vs. absence: 60 ng/mL, p < 0.0001, AUC = 0.93). FcgammaBP in amniotic fluid might be a marker of intra-amniotic infection in women with both PPROM and PTL However, in cervical fluid, it is only observed in women with PPROM.

Intra-amniotic infection and sterile intra-amniotic inflammation are associated with elevated concentrations of cervical fluid interleukin-6 in women with spontaneous preterm labor with intact membranes

Objective To determine the concentration of interleukin-6 (IL-6) in the cervical fluid in women with spontaneous preterm labor with intact fetal membranes (PTL) complicated by intra-amniotic infection (the presence of both microbial invasion of the amniotic cavity and intra-amniotic inflammation), or sterile intra-amniotic inflammation (the presence of intra-amniotic inflammation alone). Methods Eighty women with singleton pregnancies complicated by PTL between gestational ages 22 + 0 and 34 + 6 weeks were included in this retrospective cohort study. Samples of amniotic and cervical fluids were collected at the time of admission. Amniotic fluid samples were obtained via transabdominal amniocentesis, and cervical fluid was obtained using a Dacron polyester swab. Microbial invasion of the amniotic cavity was diagnosed based on the combination of culture and molecular biology methods. The concentration of IL-6 in the amniotic and cervical fluids were measured using an automated electrochemiluminescence immunoassay method. Intra-amniotic inflammation was defined as an amniotic fluid IL-6 concentration ≥3000 pg/mL. Results The presence of intra-amniotic infection and sterile inflammation was identified in 15% (12/80) and 26% (21/80) of the women, respectively. Women with intra-amniotic infection (median: 587 pg/mL; p = .01) and with sterile intra-amniotic inflammation (median: 590 pg/mL; p = .005) had higher concentrations of IL-6 in the cervical fluid than those without intra-amniotic inflammation (intra-amniotic infection: median 587 pg/mL vs. without inflammation, median: 136 pg/mL; p = .01; sterile intra-amniotic inflammation, median: 590 pg/mL vs. without inflammation, p = .005). No differences were found in the concentrations of IL-6 in the cervical fluid between women with intra-amniotic infection and sterile intra-amniotic inflammation (p = .81). Conclusion In pregnancies with PTL, both forms of intra-amniotic inflammation are associated with elevated concentrations of IL-6 in the cervical fluid.

Diagnostic Efficacy of DNA Ploidy in Liquid Based Cervical Cytology using DNA Cytometry

Worldwide cervical cancer is the fourth most common cancer in women and high incidence is reported from India. Liquid Based Cytology (LBC) provides good morphology for detection of cellular abnormalities. We, therefore, reviewed diagnostic efficacy of conventional Pap staining, flow cytometry and Human Papilloma Virus (HPV) testing in cervical pre cancer and cancer. Narrative review of cervical pre cancer and cancer candidate biomarkers including Pap staining, HPV and flow cytometry from cervical cytology fluids, is based on a detailed review of the literature. Based on the so far conducted studies, a promising conclusion can be drawn, that cytometry when coupled with HPV DNA typing or the conventional cytology gives better results as compared to that of conventional cytology or DNA cytometry alone. Liquid cytology provides a good and stable source of cervical cells to carry out ploidy studies using DNA cytometry. The procedure should be used in conjunction with LBC and HPV detection.

J-107 Interrogating the immunologic and microbial cervical microenvironment in HPV infection

Infection with high-risk human papillomavirus (hrHPV) infection in the oral and anogenital mucosa has been confirmed to be a necessary but insufficient cause of invasive cervical cancer (ICC), which is a leading cause of cancer incidence and mortality worldwide. Over the past 2 decades, our understanding of the natural history of cervical hrHPV infection has evolved, especially with regard to the influence of co-infections and the dynamics of the cervical microenvironment. For example, it is now recognized that at least a proportion of hrHPV infections are immunologically controlled for long periods of time in an undetectable (latent) state, at risk of recurrent detection (reactivation) upon immune suppression.1 This phenomenon is most clearly illustrated when comparing the HPV natural history in HIV-infected and HIV-uninfected individuals, where both incident and persistent detection of HPV is proportionally increased with increasing immune suppression.2 Multiple new type cervical HPV detection increases nearly 4-fold within weeks of acute HIV infection, suggesting a key role for tissue resident T-cell memory in control of latent HPV infection.3 In HIV-uninfected women, we have observed polarization toward T-helper type 2 immune profiles of the lower genital tract immune microenvironment. In an older, well-screened US population, HPV positive women exhibited immune profiles similar to those described in allergic inflammation.4 In the developing world, we have reported an increased prevalence of HPV infection at older ages in Amazonian women with soil-transmitted helminth (STH) infection, who showed evidence of Th2-skewed immunity in lower genital tract secretions compared with STH-uninfected women.5 This epidemiologic evidence, combined with experimental animal models of papillomavirus infection, collectively implicate a key role of host immunity in the control of cervical HPV infection and, by extension, risk of cervical cancer. This presentation will make a case that the time has come to move human HPV natural history studies away from traditional designs with large sample sizes and long interval censored sampling (eg, every 6 months) to designs with smaller sample sizes and more frequent sampling density (eg, every week) to allow interrogation of the dynamic interplay between the shifting microenvironment and HPV detection. Prioritizing a focus on understanding the dynamics and immune mediators involved in healthy control of HPV infection may be an overlooked fundamental system component required to fully understand the causes of dysregulation of HPV infection and persistent detection. This represents a paradigm shift in studying the molecular epidemiology of human viral infections, reflects more fully a systems approach to understanding biological complexity, and may lead to development of less invasive interventions at earlier stages in viral natural history to prevent severe pathological outcomes. REFERENCES 1. Gravitt PE. The known unknowns of HPV natural history. J Clin Invest. 2011;121:4593–4599. 2. Strickler HD, Burk RD, Fazzari M, et al. Natural history and possible reactivation of human papillomavirus in human immunodeficiency virus-positive women. J Natl Cancer Inst. 2005;97:577–586. 3. Nowak RG, Gravitt PE, Morrison CS, et al. Increases in human papillomavirus detection during early HIV infection among women in Zimbabwe. J Infect Dis. 2011;203:1182–1191. 4. Marks MA, Viscidi RP, Chang K, et al. Differences in the concentration and correlation of cervical immune markers among HPV positive and negative perimenopausal women. Cytokine. 2011;56:798–803. 5. Gravitt PE, Marks M, Kosek M, et al. Soil-transmitted helminth infections are associated with an increase in human papillomavirus prevalence and a T-helper type 2 cytokine signature in cervical fluids. J Infect Dis. 2016;213:723–730.

Use of Natural Family Planning (NFP) and Its Effect on Couple Relationships and Sexual Satisfaction: A Multi-Country Survey of NFP Users from US and Europe.

Birth control is a persistent global health concern. Natural family planning (NFP) comprises methods to achieve or avoid pregnancy independent of mechanical or pharmacological intervention. The sympto-thermal method (STM) of NFP employs daily observation of cervical fluids and measurement of basal body temperature. This multi-country study was undertaken to describe the characteristics of STM users, understand their perceptions of NFP, and its perceived impact on relationships. Questionnaires for women and men were developed in German and translated to English, Polish, Italian, Czech, and Slovak by native speakers. A total of 2,560 respondents completed the online questionnaire (37.4% response). Participants were married (89%) and well educated, and their self-perceived financial status was described as "good" or "very good" by 65% of the respondents. Forty-seven percent had previously used contraceptives. Ninety-five percent of women and 55% of men said using NFP has helped them to know their body better. Large majorities of men (74%) and women (64%) felt NFP helped to improve their relationship while <10% felt use of NFP had harmed their relationship. Most women (53%) and men (63%) felt using NFP improved their sex life while 32% of women and 24% of men felt it was unchanged from before they used NFP. Seventy-five percent of women and 73% of men said they are either "satisfied" or "very satisfied" with their frequency of sexual intercourse. This survey demonstrates STM of NFP is a well-accepted approach to family planning across several Western cultures. It is consistently viewed as being beneficial to couples' self-knowledge, their relationship, and satisfaction with frequency of sexual intercourse.

Cervical fluid interleukin 6 and intra-amniotic complications of preterm prelabor rupture of membranes

Objective: To determine if cervical fluid interleukin (IL)-6 concentrations in women with preterm prelabor rupture of membranes (PPROM) allows identification of microbial invasion of the amniotic cavity (MIAC) and/or intra-amniotic inflammation (IAI).Methods: One hundred forty-four women with singleton pregnancies complicated by PPROM were included in this prospective cohort study. Cervical and amniotic fluids were collected at the time of admission and concentrations of IL-6 were measured using an ELISA and point-of-care test, respectively. Cervical fluid was obtained using a Dacron polyester swab and amniotic fluid was obtained by transabdominal amniocentesis. MIAC was diagnosed based on a positive PCR result for Ureaplasma species, M. hominis, and/or C. trachomatis and/or by positivity for the 16 S rRNA gene. IAI was defined as amniotic fluid point-of-care IL-6 concentrations ≥745 pg/mL. The women were assigned to four subgroups based on the presence of MIAC and/or IAI: microbial-associated IAI (both MIAC and IAI), sterile IAI (IAI alone), MIAC alone, and without either MIAC or IAI.Results: (1) Women with microbial-associated IAI had higher cervical fluid IL-6 concentrations (median 560 pg/mL) than did women with sterile IAI (median 303 pg/mL; p = .001), women with MIAC alone (median 135 pg/mL; p = .0004), and women without MIAC and IAI (median 180 pg/mL; p = .0001). (2) No differences were found in cervical fluid IL-6 concentrations among women with sterile IAI, with MIAC alone, and without MIAC and IAI. (3) A positive correlation was observed between cervical fluid IL-6 concentrations and the amount of Ureaplasma species in amniotic fluid (copies DNA/mL; rho = 0.57, p < .0001). (4) A weak positive correlation was detected between cervical and amniotic fluid IL-6 concentrations (rho = 0.33, p < .0001).Conclusions: The presence of microbial-associated IAI is associated with the highest cervical fluid IL-6 concentrations. Cervical IL-6 can be helpful in the identification of microbial-associated IAI.

Cervical fluid calreticulin and cathepsin-G in pregnancies complicated by preterm prelabor rupture of membranes

Objective: The study aimed to determine the cervical calreticulin and cathepsin-G concentrations in pregnancies complicated by preterm prelabor rupture of membranes (PPROM) with respect to the presence of microbial invasion of the amniotic cavity (MIAC) and intra-amniotic inflammation (IAI).Methods: Eighty women with singleton pregnancies complicated by PPROM were included in this study. Cervical and amniotic fluids were obtained at the time of admission, and concentrations of calreticulin and cathepsin-G in cervical fluid were determined using ELISA. The MIAC was defined as a positive PCR analysis for Ureaplasma species, Mycoplasma hominis, and/or Chlamydia trachomatis and/or by positivity for the 16S rRNA gene. IAI was defined as amniotic fluid bedside IL-6 concentrations ≥745 pg/mLResult: Neither women with MIAC nor with IAI had different cervical fluid concentrations of calreticulin (with MIAC: median 18.9 pg/mL vs. without MIAC: median 14.7 pg/mL, p = 0.28; with IAI: median 14.3 pg/mL vs. without IAI: median 15.6 pg/mL, p = 0.57;) or of cathepsin-G (with MIAC: median 30.7 pg/mL vs. without MIAC: median 24.7 pg/mL, p = 0.28; with IAI: median 27.3 pg/mL vs. without IAI: median 25.1 pg/mL, p = 0.80) than women without those complications. No associations between amniotic fluid IL-6 concentrations, gestational age at sampling, and cervical fluid calreticulin and cathepsin-G concentrations were found.Conclusions: Cervical fluid calreticulin and cathepsin-G concentrations did not reflect the presence of MIAC or IAI in women with PPROM.

Biochemical fingerprints of human papillomavirus infection and cervical dysplasia using cervical fluids: Spectral pattern investigation.

The Pap smear is the primary screening tool for invasive cervical cancer resulting from a persistent infection with oncogenic human papillomavirus (HPV); however, there are the problems such as the inability to distinguish between HPV infection and cervical dysplasia and a low sensitivity remain. We present preliminary findings of a label-free method to detect and classify HPV infection and cervical dysplasia using human cervical fluids. Three experimental groups, defined as normal, HPV-positive, and cervical dysplasia, were evaluated through their Raman spectral patterns for noise-independence, high reproducibility, and uniformity. Clinical diagnosis was performed through liquid-based cervical cytology, HPV test, and cervical histologic examination. Healthy cervical fluids showed a strong Raman intensity at 877 cm-1 (symmetric C-C stretching), and at 963 cm-1 (phosphate), compared to a reference Raman peak at 1003 cm-1 (phenylalanine symmetric ring breath). The HPV-positive cervical fluids showed a strong intensity of a Raman peak at 1448 cm-1 corresponding to C-H deformation vibration mode and the highest similarity between the central and ring zones among the three groups. The cervical dysplasia fluids showed the presence of strong peaks compared to the control and HPV-positive groups. In addition, different Raman spectra were acquired according to HPV type. Therefore, all ranges of cervical fluid-induced Raman spectra could be used to detect the presence of cervical pre-cancer. Raman peak-gated assessment provides a label-free and nondestructive tool for the clinical diagnosis of HPV infection and cervical precancerous changes.

Soil-Transmitted Helminth Infections Are Associated With an Increase in Human Papillomavirus Prevalence and a T-Helper Type 2 Cytokine Signature in Cervical Fluids.

An ecological correlation between invasive cervical cancer incidence and burden of soil-transmitted helminths (STH) is hypothesized to explain the excess in detectable human papillomavirus (HPV) infection in Latin America, via a global T-helper type 2 (Th2)-biased mucosal immune response secondary to STH infection. The association between current STH infection and HPV prevalence was compared in regions of Peru where STH is or is not endemic. Adjusted prevalence ratios (PRs) with robust variance were estimated as an effect measure of STH infection on HPV prevalence in each study site. Soluble immune marker profiles in STH-infected and STH-uninfected women were compared using Spearman rank correlation with the Sidak correction. Among women in the helminth-endemic region of the Peruvian Amazon, those with STH infection women had a 60% higher prevalence of HPV, compared with those without STH infection (PR, 1.6; 95% confidence interval, 1.0-2.7). Non-STH parasitic/protozoal infections in the non-STH-endemic population of Peru were not associated with HPV prevalence. In Iquitos, A Th2 immune profile was observed in cervical fluid from helminth-infected women but not helminth-uninfected women. A proportion of the increased HPV prevalence at older ages observed in Latin America may be due to a population-level difference in the efficiency of immunological control of HPV across the lifespan due to endemic STH infection.

Biochemical investigations of human papillomavirus-infected cervical fluids.

Cervical cancer is caused by persistent infection with high-risk types of human papillomavirus (HPV). The aim of this study was to investigate the potential of Raman spectroscopy as a stand-alone analytical tool for clinical diagnosis of HPV infection using human cervical fluids. A drop-coating deposition surface-enhanced Raman scattering (DCD-SERS) method was identified as the most effective method of biochemical analysis in cervical biofluids. Using a 2-µL sample, the proposed DCD-SERS method yielded Raman spectra with high reproducibility, noise-independence, and uniformity. Additionally, the produced spectra were independent of the volume of fluid used and detection zone analyzed within the central and the ring zones. The optical detection of HPV infection in cervical fluids could be detected accurately in the central zone. Different Raman spectra were obtained according to HPV type. In particular, HPV-16 and HPV-18, which are the major risk factors for cervical cancer, showed a distinct spectral difference in cervical fluids, with a similar pattern for high Raman bands at >1,000 cm(-1) but a different pattern for low Raman bands at <1,000 cm(-1) . Therefore, all ranges of HPV-gated DCD-SERS could be used to detect the presence of HPV infection. Raman spectroscopy provides a good alternative method for early clinical diagnosis of HPV infection, and we are hopeful that this proposed method of HPV assessment will be approved for use in obstetrics and gynecology clinics.

Progesterone and threatened abortion: a randomized clinical trial on endocervical cytokine concentrations

The purpose of this study was to investigate the effect of vaginal progesterone on endocervical cytokine concentration in women at risk of threatened abortion. One hundred and sixty pregnant women with clinical symptoms of threatened abortion before the 20th week of pregnancy were randomly assigned to receive vaginal progesterone or placebo. Cervical fluids were collected and endocervical concentrations of different cytokines (IFNγ, TNFα, IL-8, IL-10 and IL-12) were analyzed before and one week after progesterone or placebo treatment. A significant decrease in IFNγ and increase in IL-10 in endocervical fluid was seen when the values were compared before and after progesterone treatment. However, there were no significant differences in pregnancy outcomes between the placebo and progesterone groups. We conclude that despite the failure of vaginal progesterone treatment to improve pregnancy outcomes, progesterone can induce a shift in the concentration of cytokines in endocervical secretions.

A clinical randomized trial on endocervical inflammatory cytokines and betamethasone in prime-gravid pregnant women at risk of preterm labor.

There are strong evidences suggesting the secretion of different cytokines in cervical fluid during preterm labor. Betamethasone is widely administered for several reasons in preterm conditions. To Investigate the possible effect of betamethasone on endocervical cytokine concentration of women at risk of preterm labor. In a randomized clinical trial of 80 prime-gravid women in preterm labor between 34 and 37 weeks of gestation, cervical fluids were collected. Endocervical concentration of inflammatory cytokines were analyzed before and 48 hours after betamethasone treatment for the evaluation of IL-8, IL-17, IFN-γ and TGF-β. Wilcoxon and Mann-Whitney tests were employed for statistical analysis. χ2 and Student's t tests were used whenever needed. All the measured cytokines showed significant changes in the betamethasone treated group. IL-17 (p=0.001), IL-8 (p=0.001), and IFN-γ (p<0.05) decreased significantly, while TGF-β had a significant increase (p<0.05). In the patients who delivered before or on the 7th day of admission, IL-17, IL-8, and IFN-γ levels were all significantly higher. However, TGF-β decreased significantly in the same samples in the betamethasone treated group (p<0.05). Betamethasone significantly decreases the endocervical pro-inflammatory cytokine concentrations in patients with preterm labor.

PD 404,182 Is a Virocidal Small Molecule That Disrupts Hepatitis C Virus and Human Immunodeficiency Virus

We describe a virucidal small molecule, PD 404,182, that is effective against hepatitis C virus (HCV) and human immunodeficiency virus (HIV). The median 50% inhibitory concentrations (IC(50)s) for the antiviral effect of PD 404,182 against HCV and HIV in cell culture are 11 and 1 μM, respectively. The antiviral activity of PD 404,182 is due to the physical disruption of virions that is accompanied to various degrees (depending on the virus and exposure temperature/time) by the release of viral nucleic acids into the surrounding medium. PD 404,182 does not directly lyse liposomal membranes even after extended exposure, and it shows no attenuation in antiviral activity when preincubated with liposomes of various lipid compositions, suggesting that the compound inactivates viruses through interaction with a nonlipid structural component of the virus. The virucidal activity of PD 404,182 appears to be virus specific, as little to no viral inactivation was detected with the enveloped Dengue and Sindbis viruses. PD 404,182 effectively inactivates a broad range of primary isolates of HIV-1 as well as HIV-2 and simian immunodeficiency virus (SIV), and it does not exhibit significant cytotoxicity with multiple human cell lines in vitro (50% cytotoxic concentration, >300 μM). The compound is fully active in cervical fluids, although it exhibits decreased potency in the presence of human serum, retains its full antiviral potency for 8 h when in contact with cells, and is effective against both cell-free and cell-associated HIV. These qualities make PD 404,182 an attractive candidate anti-HIV microbicide for the prevention of HIV transmission through sexual intercourse.

Prediction of microbial invasion of the amniotic cavity in women with preterm labour: analysis of multiple proteins in amniotic and cervical fluids

Microbial invasion of the amniotic cavity is a major cause of preterm delivery and the diagnosis is dependent on invasive amniocentesis. The objective was to determine whether specific proteins in amniotic and cervical fluids alone, or in combination, could identify bacterial invasion. A prospective follow-up study. Women with singleton pregnancies presenting with preterm labour between 22 and 33 weeks of gestation (n = 89). Sahlgrenska University Hospital, Gothenburg, Sweden. Amniotic and cervical fluid was analysed with polymerase chain reaction for Mycoplasmas, and was cultured for aerobic and anaerobic bacteria. Twenty-seven proteins were analysed using multiplex technology. Individual levels of each protein were compared in order to find associations between different proteins and microbial invasion of the amniotic cavity. Predictive models based on multiple proteins were created using stepwise binary logistic regression. The main outcome measure was microbial invasion of the amniotic cavity. Microbial invasion of the amniotic cavity was present in 17% (15/89) of the women. Concentration levels of several amniotic and cervical proteins were significantly higher in women with microbial invasion of the amniotic cavity. Three multivariate predictive models were found. The predictive power of the non-invasive model (73% sensitivity, 88% specificity, 55% positive predictive value, 94% negative predictive value) was as good as the invasive models. Area under the receiver operating characteristic (ROC) curve and likelihood ratio were 0.87 and 6.0, respectively. Prediction of intra-amniotic infection using selected cervical proteins was equally good as prediction using the same proteins collected from amniotic fluid, or a combination of cervical and amniotic proteins.

Factors affecting matrix metalloproteinase-8 levels in the vaginal and cervical fluids in the first and second trimester of pregnancy

Cervical ripening during pregnancy resembles an inflammatory process. Matrix metalloproteinases (MMPs), particularly MMP-8, have been linked to inflammatory processes. We studied the concentrations of, and factors associated with, MMP-8 in the lower genital tract fluids in the first and second trimesters. In a prospective population-based cohort study, vaginal and cervical swab samples were obtained from 2130 unselected pregnant women undergoing their first and second trimester ultrasound screening. MMP-8 was determined by immunofluorometric assay. Use of antibiotics, history of vaginal bleeding, and history of sexual intercourse were recorded on both occasions. Vaginal smears were obtained for Gram-staining and leukocyte counting. Cervical length was measured by ultrasonography. The main outcome measures were MMP-8 concentrations in the vagina and cervix. The median (range) MMP-8 concentrations in vaginal and cervical samples were 107.4 microg/l (undetectable-2406.6 microg/l) and 318.3 microg/l (0.1-2074.6 microg/l), respectively, in the first trimester, and 112.5 microg/l (undetectable-2093.4 microg/l) and 344.8 microg/l (0.4-1783.5 microg/l), respectively, in the second trimester. Multiparity and vaginal leukocytosis were both associated with increased MMP-8 concentrations in vaginal and cervical samples in both trimesters. Bacterial vaginosis (BV) was associated with increased vaginal and cervical MMP-8 in the first trimester, but only with increased vaginal MMP-8 in the second trimester. A history of sexual intercourse (in the previous 48 h) was associated with lower MMP-8 concentrations in cervical samples in both trimesters. MMP-8 concentrations were lower in vaginal samples than in cervical samples, and no difference was found between the first and second trimesters. Multiparity, BV and an elevated leukocyte count in the vagina were associated with increased MMP-8 concentrations. Sexual intercourse had an opposite effect. The study suggests that MMP-8 is a physiologic constituent in lower genital tract fluids, where it may be involved in host response to inflammatory and infectious processes.

Prediction of Spontaneous Preterm Delivery in Women With Preterm Labor

To analyze whether specific proteins in amniotic and cervical fluids, alone or in combination with risk factors, can identify women in preterm labor with intact membranes who will deliver spontaneously within 7 days of sampling. In a cohort of 89 women in preterm labor, amniotic and cervical fluids were collected between 22 and 33 weeks of gestation. Twenty-seven proteins were analyzed simultaneously using multiplex technology. Individual levels of each protein were compared and calculations performed to find associations among different proteins, background variables, and spontaneous preterm delivery within 7 days of sampling. The area under the curve (AUC) was calculated using receiver operating characteristic curve analysis, and prediction models were created based on stepwise logistic regression. We found two multivariable models that predicted spontaneous preterm delivery better than any single variable. One combined multivariable prediction model was based on amniotic macrophage inflammatory protein-1beta, cervical interferon-gamma, and monocyte chemotactic protein-1. This model predicted outcome with 91% sensitivity, 84% specificity, 78% positive predictive value, and 94% negative predictive value, with a likelihood ratio of 5.6 and AUC of 0.91. An alternative, noninvasive model based on cervical length, cervical interferon-gamma, interleukin-6, and monocyte chemotactic protein-1 predicted delivery within 7 days with 85% sensitivity, 82% specificity, 74% positive predictive value, and 90% negative predictive value, with a likelihood ratio of 4.7 and AUC of 0.91. A combination of proteins from amniotic fluid and cervical fluid or cervical length can help determine which women will deliver preterm. II.