Cervical Fluid Research Articles

Fertility awareness-based methods of family planning.

Fertility awareness-based methods (FABMs) of family planning involve monitoring various signs and symptoms of fertility during the menstrual cycle to identify the "fertile window," or the days of the cycle when unprotected intercourse is most likely to result in pregnancy. Signs and symptoms include menstrual cycle length, basal body temperature, urinary hormone measurements, and/or cervical fluid and may be used alone or in combination. Fertility signs reflect both physiological changes during the menstrual cycle and the life cycle of the ovum and sperm. Women learn to observe or measure and interpret these signs according to the instructions of their chosen FABM and avoid unprotected intercourse on fertile days. FABMs are appropriate for those who choose to use them, are able and willing to observe one or more fertility signs, and are in relationships that support the use of a coitus-related method such as a condom or abstaining from intercourse on fertile days.

220 Exploring the use of mesenchymal stem cells for treatment of mastitis and metritis in cattle

The present study was carried out to isolate mesenchymal stem cells (MSCs) from adipose tissue of cattle (Bos indicus), characterise them, and apply them for the treatment of mastitis and metritis in the cow. Cattle MSCs were isolated from adipose tissue near the loin region of cow. Isolated adipose tissue was subjected to enzymatic digestion using 2% collagenase with agitation at regular intervals. The cells obtained after digestion were resuspended in cell culture flasks containing growth enriched medium and cultured under standard culture conditions. Alkaline phosphatase staining was used as one of the parameters to confirm cultured putative MSCs. Bovine Ad-MSCs were further characterised using real time-PCR by amplification of MSC-specific markers: CD73, CD90, and CD105 as positive markers and CD34, CD45, and CD79a as negative markers. Immunocytochemistry showed the presence of CD73, CD90, and CD105 on the cell surface. Three groups-control (C), local (L), and intravenous (IV)-with 6 cows suffering from mastitis were taken in each group and subjected to MSC transplantation through local and intravenous routes. Control group animals were subjected to antibiotic treatment only. Similarly, another three groups were taken with 6 cows in each group suffering from metritis. Post-transplantation wound healing, tissue repair, and reduction in inflammation were monitored for 26 days, at different time intervals; that is, after Days 1, 3, 7, and 15. Blood samples were also collected from animals at the same time intervals for real time-PCR. A similar examination was also done in metritis groups along with the analysis of the reduction in turbidity of cervical fluid at the abovementioned time intervals. Real time-PCR was performed to determine relative expression of genes for proliferative factors, anti-inflammatory cytokines, and antimicrobial peptides on cells isolated from blood collected at different time intervals. Gene expression in the local group of mastitis subjected to MSC injection was significantly higher than that of the IV and control group. The somatic cell count declined in both local and IV groups compared with the control group. Whereas the expression of the same genes in the IV group of metritis was significantly higher than that of the local and control groups of cows. The turbidity of cervical fluid and mucus was reduced in the IV group compared with the local group. In conclusion, we demonstrated the healing potential of MSCs in a cow model via MSC injection. Promising results were obtained in curing mastitis in both local and IV groups, whereas healing in the case of metritis was significantly higher in the IV group compared with both the control and local groups of cows. The study indicates the potential use of MSc for treatment of mastitis and metritis in cattle through wound healing and decreasing microbial infection.

Molecular detection of HPV and FT-IR spectroscopy analysis in women with normal cervical cytology.

The increase in the incidence of Cervical Cancer in the female population worldwide has been an issue that deserves further attention from the scientific community. Several studies have already proven the relationship of its development with the molecular mechanisms that Human Papillomavirus (HPV) induces in cervical cells. The gene amplification provided by molecular biology techniques has been used as the gold standard diagnostic method of this virus because of its high specificity and sensitivity.However, the high investments associated with the acquisition of reagents, equipment and labor demonstrate the need for the development of more accessible techniques that present the same accuracy. FT-IR spectroscopy has been studied as an inexpensive and easily accessible technology that can provide the differentiation of malignant and benign cells. This study aimed to demonstrate the effectiveness and sensitivity of molecular analysis by PCR in relation to cytological analysis and to evaluate the sensitivity of FT-IR spectroscopy in the diagnosis of HPV for cervical cancer prevention. Cervical fluid samples obtained from 50 patients with absence of cellular lesion by cytological analysis were analyzed by molecular and spectroscopic analyzes. Oncotic colpocitology analysis was performed by the Papanicolaou staining, amplification of the L1 viral gene by PCR was performed using primers MY09 and MY11 and biochemical analysis of the fluids by FT-IR was performed using the Spectrum 400 system equipped with a microscope. Of the 50 patients without evident morphological alteration of the cells, seven were diagnosed by molecular analysis as positive for presence of HPV. Principal component analysis of spectroscopy was not able to separate the negative samples from the HPV positive samples and, therefore, did not present as an effective diagnostic technique. We highlight the efficacy, sensitivity and specificity of molecular biology by PCR in the identification of the virus and we emphasize that more studies should be used for the application of FT-IR spectroscopy in the diagnosis of this infection and its application in the prevention of cervical cancer.

Fecundability in relation to use of fertility awareness indicators in a North American preconception cohort study

To quantify the frequency of use of selected fertility awareness indicators and to assess their influence on fecundability. Web-based prospective cohort study. Not applicable. Female pregnancy planners, aged 21-45years, attempting conception for ≤6 cycles at study entry. None. We ascertained time to pregnancy, in menstrual cycles, with bimonthly questionnaires. We estimated adjusted fecundability ratios (FRs) and confidence intervals (CIs) using proportional probabilities models, controlling for age, income, education, smoking, intercourse frequency, and other lifestyle and reproductive factors. A total of 5,688 women were analyzed, with a mean age of 29.9years and mean time trying of 2.1 cycles at baseline; 30% had ever been pregnant. At baseline, 75% were using one or more fertility indicators (counting days or charting menstrual cycles [71%], measuring basal body temperature [BBT, 21%], monitoring cervical fluid [39%], using urine LH tests [32%], or feeling for changes in position of the cervix [12%]). Women using any fertility indicator at baseline had higher subsequent fecundability (adjusted FR 1.25, 95% CI 1.16-1.35) than those not using any fertility indicators. For each individual indicator, adjusted FRs ranged from 1.28-1.36, where 1.00 would indicate no relation with fecundability. The adjusted FR for women using a combination of charting days, cervical fluid, and urine LH was 1.48 (95% CI 1.31-1.67) relative to women using no fertility indicators. In a North American preconception cohort study, use of fertility indicators indicating the fertile window was common, and was associated with greater fecundability.

2484 Clinical Outcomes Among Women with Abnormal Uterine Bleeding Treated with Inpatient or Outpatient Hysterectomy Versus Endometrial Ablation

Study Objective To describe clinical and economic outcomes among women with abnormal uterine bleeding (AUB) treated with global endometrial ablation (GEA) versus inpatient and outpatient hysterectomy. Design US retrospective claims analysis. Setting N/A. Patients or Participants Women undergoing GEA or hysterectomy (index event) between 1/1/2012 and 4/30/2017 with ≥2 non-diagnostic claims indicating AUB before or on the index date in the IBM MarketScan Commercial Database. Women were required to have 12 months pre- and post-index continuous enrollment. Women with evidence of pregnancy, delivery, menopause, non-skin cancer or receiving total abdominal hysterectomy were excluded. Follow-up healthcare costs and the incidence of complications (such as cervical occlusion, cervical/uterine/bowel perforation, fluid overload, pyometra, cervicitis, device complications and cervical trauma) were reported. Re-intervention, defined as hysterectomy or repeat GEA, was reported during the five-year post-index among GEA patients. Interventions GEA or hysterectomy. Measurements and Main Results 117,801 women met the study criteria (56.6% GEA, 43.4% hysterectomy). 84.0% of hysterectomy cases occurred in an outpatient setting. Mean age was lower for GEA patients (42.7 vs. 43.2). Total healthcare costs in the first month post-index among GEA patients were less than one-half of hysterectomy ($7,018 vs. $15,402), a similar finding when limited to inpatient ($17,672) or outpatient hysterectomy ($14,971) (all p Conclusion This analysis of a large, national claims database found that GEA was approximately one-half the cost of hysterectomy (inpatient or outpatient) for the treatment of AUB with fewer complications and declining rates of re-intervention.

DNA-HPV transition rate and related factors in HPV-infected women in Can Tho city, Vietnam.

To determine DNA-HPV transition rates and related factors in HPV-infected women 18-69years of age in Can Tho City from 2013 to 2018. Both a retrospective and a prospective cohort study were done. Interviews, gynaecological examinations and HPV testing by PCR (cervical fluid) were used to collect data. The results were recorded and compared with those of HPV in 2013 to assess the development of HPV over time. Transition was defined as conversion to HPV-positive state in 2018 from a negative state in 2013. No transition was defined as clearance of HPV when the positive 2013 result was negative in 2018 or when the result remained negative or positive in 2013 and 2018. Factors related to the change were analysed. Among a sample size of 204 cases, the average age of participants was 48.9±10.4years. Women >45 comprised 63.2% of participants; 82.8% lived with their husbands, 6.4% were divorced, and 2.9% lived apart from their husbands due to work. After 5years of observation, 16.2% of DNA-HPV cases had converted to HPV-positive state and 66.2% of DNA-HPV cases had cleared to HPV-negative state. Factors related to conversion to HPV-positive state were age≤45years (3.14 times higher risk of transition than in the >45 age group (95% CI: 1.12-8.8)); change of sexual partner (OR=3.75 (95% CI: 1.15-12.2)); change of sexual partner by husband (OR=3.69 (95% CI: 1.20-11.3); sexually transmitted diseases (OR=5.19 (95% CI: 1.09-24.8)); and a history of vacuum aspiration or dilation and evacuation abortion (OR=1.4 (95% CI: 0.29-6.4)). 16.2% of women with DNA-HPV transition converted to HPV-positive state. Changes in sexual habits increase the risk of developing HPV positivity.

Predictive value of cervical cytokine, antimicrobial and microflora levels for pre-term birth in high-risk women

Spontaneous preterm birth (sPTB, delivery <37 weeks gestation), accounts for approximately 10% of births worldwide; the aetiology is multifactorial with intra-amniotic infection being one contributing factor. This study aimed to determine whether asymptomatic women with a history of sPTB or cervical surgery have altered levels of inflammatory/antimicrobial mediators and/or microflora within cervical fluid at 22–24 weeks gestation. External cervical fluid was collected from women with history of previous sPTB and/or cervical surgery at 22–24 weeks gestation (n = 135). Cytokine and antimicrobial peptides were measured on a multiplex platform or by ELISA. qPCR was performed for detection of 7 potentially pathogenic bacterial species. IL-8 and IL-1β levels were lower in women who delivered preterm compared to those who delivered at term (IL-8 P = 0.02; IL-1β P = 0.04). There were no differences in elafin or human beta defensin-1 protein levels between the two groups. Multiple bacterial species were detected in a higher proportion of women who delivered preterm than in those who delivered at term (P = 0.005). Cervical fluid IL-8 and IL-1β and microflora have the potential to be used as biomarkers to predict sPTB in high risk women.

Acute Longus Colli Calcific Tendinitis: A Rare Cause of Neck Pain.

A 48-year-old man presented to our emergency department with neck pain without sensorimotor deficit and with a sore throat without signs of infection. Magnetic resonance imaging was performed because the patient had not responded to regular treatment and a blood test had showed inflammation. The images revealed cervical prevertebral fluid collection and calcification, compatible with acute calcific tendinitis of the longus colli muscle. Prednisolone 50 mg with pantoprazol 40 mg was administered for 5 days with rapid resolution of symptoms. Acute calcific tendinitis of the longus colli muscle is a rare and possibly underdiagnosed cause of atypical neck pain and sore throat.LEARNING POINTSThe signs and symptoms of acute calcific tendinitis of the longus colli muscle can easily be mistaken for serious disease such as spondylodiscitis or retropharyngeal abscess.Imaging with easily available modalities such as computed tomography can be used for diagnosis.Anti-inflammatory medication led to quick resolution of symptoms despite elevated signs of inflammation and delayed diagnosis in our patient.

Preterm Birth Prediction by Measurement of Biomarkers in Cervical Vaginal Fluid

Preterm Birth Prediction by Measurement of Biomarkers in Cervical Vaginal Fluid

Compliance with daily, home-based collection of urinary biospecimens in a prospective, preconception cohort.

Exposures in the periconceptional environment may impact fertility and future health. Assessing time-varying exposures during the periconceptional window requires identifying approximate fertile windows around ovulation. In this prospective cohort study, we instructed women in daily cervical fluid observation and interpretation to identify incipient ovulation; they used this information to time daily urine collection for both partners. Timing and completeness of collection were compared to expert review. One hundred seventy couples planning pregnancy enrolled from community volunteers from 2011 to 2015; women were taught the Peak Day method to identify fertile windows. Both partners collected daily urine specimens from the first day of fertile-quality fluid (estimator of the beginning of fertile window). Men discontinued on the estimated day of ovulation/conception +2 days; women continued through the onset of next menses, or positive pregnancy test at estimated day of ovulation/conception +18 days. We compared dates from samples with participants' fertility charts to determine proportion correctly collected. Also, expert reviewers judged on which days urine should have been collected, determining investigator-identified sampling days. One hundred sixty-nine couples submitted 6,118 urine samples from 284 cycles. Reviewers and participants agreed in 87% of cycles for the date of the beginning of the fertile window ±3 days (65% exact-day agreement); agreement on ovulation date, ±3 days, was 93% (75% exact-day agreement). Five thousand three hundred twenty-nine female samples were expected based on investigator-identified sampling days, and 4,546 were collected, of which 82% were correctly collected on expected days. Fifty-nine percent of male samples were correctly collected relative to investigator-identified sampling days. Intensively-scheduled, biologically-triggered, at-home biospecimen collection can successfully be targeted to the periconceptional window and completed in a longitudinal cohort study.

Nicotinamide phosphoribosyltransferase and intra-amniotic inflammation in preterm prelabor rupture of membranes

Introduction: The amniotic fluid nicotinamide phosphoribosyltransferase (NAMPT) levels have not been compared among women with preterm prelabor rupture of membranes (PPROM) comorbid with intra-amniotic infection, sterile intra-amniotic inflammation (IAI), colonization, or without IAI and microbial invasion of the amniotic cavity (MIAC). Therefore, the main aim was to quantify the amniotic fluid NAMPT in women with PPROM complicated by intra-amniotic infection, sterile IAI, or colonization. The second aim was to characterize the diagnostic indices of NAMPT to reveal IAI. The third aim was to determine whether the cervical fluid and maternal serum NAMPT quantitation might be of value in the identification of intra-amniotic inflammatory complications in PPROM.Methods of study: NAMPT levels in amniotic fluid, cervical fluid, and maternal serum were assessed in three independent cohorts of women with singleton pregnancies complicated by PPROM between 24+0 and 36+6 weeks of gestation consisting of 88, 121, and 88 women, respectively. Amniotic fluid samples were obtained by transabdominal amniocentesis, cervical fluid samples were obtained using a Dacron polyester swab and maternal blood was obtained by venipuncture of the cubital vein. The NAMPT levels were measured by an enzyme-linked immunosorbent assay. Testing for MIAC and IAI was performed on all women, who were then categorized into four subgroups: intra-amniotic infection (MIAC and IAI), sterile IAI (IAI alone), colonization (MIAC alone), and without MIAC and IAI.Results: Women with intra-amniotic infection and women with sterile IAI had higher NAMPT levels than did women with colonization and women without MIAC and IAI (intra-amniotic infection: median 73.6 ng/mL, sterile IAI: median 55.5 ng/mL, colonization: median 12.1 ng/mL, without MIAC and IAI: 10.6 ng/mL; p < .0001). An amniotic fluid NAMPT level of 37 ng/mL was the best value for the detection of intra-amniotic infection in women with PPROM. Cervical fluid (p = .51) and maternal serum (p = .50) NAMPT levels did not reflect intra-amniotic inflammatory complications in women with PPROM.Conclusions: Intra-amniotic infection and sterile IAI are associated with higher NAMPT levels in amniotic fluid but not in cervical fluid or maternal serum in women with PPROM. Amniotic fluid NAMPT might be a marker for invasive identification of IAI in PPROM.

Pentraxin 3 in Noninvasively Obtained Cervical Fluid Samples from Pregnancies Complicated by Preterm Prelabor Rupture of Membranes

Problem: To determine the changes of pentraxin 3 (PTX3) level in noninvasively obtained cervical fluid samples from women with preterm prelabor rupture of membranes (PPROM) based on the presence of microbial invasion of the amniotic cavity (MIAC) and/or intra-amniotic inflammation (IAI), and intra-amniotic infection (the presence of both MIAC and IAI). Methods of Study: A total of 160 women with PPROM were included. Cervical fluid samples were obtained using a Dacron polyester swab and amniotic fluid samples were obtained by transabdominal amniocentesis. Cervical fluid PTX3 levels were assessed using enzyme-linked immunosorbent assay. Results: PTX3 was found in all the cervical fluid samples and its levels were higher in women with MIAC, IAI, and intra-amniotic infection than in women without these conditions. When the women were categorized into four subgroups based on the presence of MIAC and/or IAI, women with intra-amniotic infection had higher cervical fluid PTX3 levels than those with sterile IAI (IAI alone), colonization (MIAC alone), or no MIAC or IAI. A cervical fluid PTX3 level of 11 ng/mL was the best value for identifying the presence of intra-amniotic infection in women with PPROM. Conclusions: PTX3 is a constituent of cervical fluid of women with PPROM. Cervical fluid PTX3 level reflects the situation in the intra-amniotic compartments of women with PPROM. Cervical fluid PTX3 is a potential marker for the noninvasive identification of intra-amniotic infection in PPROM.

Influence of respiration-induced B0 variations in real-time phase-contrast echo planar imaging of the cervical cerebrospinal fluid.

Respiration induces temporal variations of the main magnetic field B0 along the spinal cord. These variations are typically not compensated for in velocity quantifications using phase-contrast MRI. The goal of this study was to analyze errors caused by respiration-induced B0 variations in real-time phase-contrast echo planar imaging (PCEPI) of cervical cerebrospinal fluid (CSF) velocity measurements and to evaluate this effect for various sequence parameters using numerical simulations. Real-time B0 measurements with double gradient echo sequence and PCEPI measurements were acquired in the cervical CSF of 10 healthy subjects. Dynamic phase offsets attributed to respiration-induced B0 variations were analyzed by quantifying amplitudes and comparing the temporal behavior with respiratory signals. In experiments and simulations, the influence of the echo time (TE) and the delay between PCEPI images (Δt) with respect to respiration on the dynamic phase offsets were investigated. A good agreement was found between phase offsets extracted from both acquisition types. Furthermore, respiratory signals qualitatively matched the temporal behavior of the measured phase offsets showing a dependency on subject-dependent local B0 distribution and respiration physiology. Simulations revealed residual background phases in PCEPI velocity quantification varying with TE and Δt. Respiration-induced B0 variations result in dynamic background phases in real-time PCEPI velocity quantifications of the CSF in the cervical spine. The current work underlines that these background phases need to be corrected to avoid confounding effects.

Amylase level in cervical drain fluid and anastomotic leakage after cervical oesophagogastrostomy.

Anastomotic leakage after oesophageal cancer surgery is a serious complication. The purpose of this study was to evaluate the possibility of anastomotic leakage by repeatedly measuring amylase levels in the fluid obtained from the drainage tube inserted at the cervical anastomotic site. Ninety-nine patients who underwent oesophagectomy and cervical oesophagogastrostomy between April 2014 and March 2017 were retrospectively reviewed. A drainage tube was placed at the anastomotic site, and amylase levels were measured daily from postoperative day (POD) 1 until oral feeding or confirmation of anastomotic leakage. The amylase levels were analysed with a linear mixed model. The mean age of the patients was 64.9 ± 9.0 years, and there were 89 (89%) male patients. Almost all pathologies (92%) were squamous cell carcinomas. The anastomotic methods were as follows: 63 (63%) circular stapled, 33 (33%) hand-sewn and 3 (3%) semistapled. Anastomotic leakage was confirmed in 10 (10%) patients. The amylase levels increased until POD 2 in both the leakage and non-leakage groups, but the levels subsequently decreased in the non-leakage group, whereas the levels peaked on POD 3 in the leakage group. On performing the linear mixed model analysis, anastomotic leakage was significantly associated with the trends in postoperative amylase levels in the drainage tube (P < 0.001). Trends in the serum C-reactive protein levels and white blood cell count were not significantly associated with anastomotic leakage. Amylase level trends measured in the cervical drain fluid can be a useful indicator of anastomotic leakage after cervical oesophagogastrostomy.

Analytical and numerical study of creeping flow generated by active spermatozoa bounded within a declined passive tract

The fluid-swimmer interaction is effective in two ways, i.e. the rheology of the surrounding fluid assists/resists the swimming mechanism and an efficient swimming generates an enhanced fluid flow. In particular the swimming of spermatozoa consists in biorheological hydrodynamic propulsion at low Reynolds numbers, where the viscous damping by far dominates over inertia. In this paper, we study the self-propulsion of a spermatozoon in the human cervix. The organism is modeled as a self-propelling infinite sheet and the cervix is simulated as a two-dimensional channel composed of two rigid walls. We assumed that the cervical walls are declined at a certain angle to the horizontal. The cervical fluid is characterized by the non-Newtonian Carreau model which exhibits shear rate-dependent viscosity, a characteristic of cervical medium. The amplitude of the waves traveling down the declined sheet surface is arbitrary while their wavelength is assumed larger than the declined channel spacing. The flow equations are formulated under this assumption. The analytic expressions of pressure gradient and velocity of the fluid are obtained by utilizing the perturbation technique. These expressions (effective for small values of Carreau parameters) are used in dynamics equilibrium conditions to calculate cell speed and power losses. The propulsive speed and power delivered by the spermatozoa at larger values of the rheological parameters are computed by employing the implicit finite deference technique combined with the Broyden method (modified secant method). The most striking result of the present study is that the optimum speed can be achieved by suitably adjusting the rheological parameters of the cervical fluid and likewise a fast moving organism can generate a better fluid flow.

1009: Smart Diaphragm Study: Multi-omics profiling and cervical device measurements during pregnancy

Examine the relationship between parturition and maternal inflammation, metabolome, proteome, vaginal microbiome, and mechanobiological changes in the cervix during pregnancy. Pregnant women were recruited from the University of California, San Francisco Hospital. Urine and cervical fluid were collected up to 15 time points, while blood samples were collected at three time points during pregnancy. Deep molecular profiling was performed at the multi-omics level, including immune panels, metabolomics, lipidomics, proteomics, and vaginal microbiome. During the same visits that the samples were collected for the multi -omics studies, a unique device called the Smart Diaphragm was used to measure fluorescence and electrical impedance of the cervical tissue. Of the 36 women who completed the study, 34 women had a delivery >37 weeks. A total of 87 blood samples, 327 urine, and 332 swab samples were collected. Using untargeted metabolomics, over 10,000 unique metabolic features were detected, in which 245 pregnancy-related metabolic features were identified in the first set of 16 women (FDR < 0.05). Pathway analysis revealed the top altered pathway in pregnancy was a steroid hormone biosynthesis pathway centered on progesterone. In addition, other altered pathways were also discovered, including the primary bile acid biosynthesis pathway. We observed dynamic changes in the biomechanical properties of the cervical tissue properties during pregnancy (12,013 fluorescence (across 17 wavelengths) and 18,252 impedance measurements (over 10 channels)). In 6 subjects, signal across all spectra decreased dramatically toward the end of the pregnancy. In other subjects, the signal was mostly stable except for some variable changes in early pregnancy. Interestingly, we found five subsets of impedance behaviors, including consistently low impedance (n= 6), spikes immediately prior to labor (n=8), spikes mid-pregnancy and immediately prior to labor (n=7), spikes early in pregnancy (n=5), and consistent spikes (n=9). In this study, we performed multi-omics profiling on samples of 36 women seen at UCSF during their pregnancies. The complete multi-omics profiles and cervical device data analysis reveal novel signatures associated with parturition, which will help explore physiological changes in pregnancy.

19: Increased concentration of 17-alpha hydroxyprogesterone caproate correlates with IL-10 to reduce spontaneous preterm birth

17-alpha hydroxyprogesterone caproate (17-OHPC) 250 mg reduces recurrent spontaneous preterm birth in women by 33%. We have previously demonstrated that women with the lowest quartile of plasma concentrations of 17- OHPC have elevated rates of preterm birth compared with women in the 2-4th quartiles. The mechanism of action of 17-OHPC is unknown but immune modulation is a possibility. We have previously reported elevated cytokines in the cervical fluid of women with a prior preterm birth. Additionally, circulating cytokines of immunoinhibitory IL-10 and pro-inflammatory TNF alpha have correlated with spontaneous preterm birth. This study sought to determine the relationship between the concentration of 17-OHPC and serum cytokines to determine if efficacy of 17-OHPC correlates with altered immunity. This was an ancillary study analyzing laboratory results from the placebo controlled, randomized trial of omega-3 supplementation and reduction of recurrent preterm birth. Peripheral blood mononuclear cells were isolated from paired blood samples drawn at 16-22 weeks gestation (baseline) and 25-28 weeks gestation (post treatment). IL-10 and TNF-alpha levels were measured with and without stimulation by lipopolysaccharide by ELISA. All women received 17-OHPC in this trial. 17-OHPC concentrations were determined from a single blood sample obtained between 25-28 weeks. Baseline and post-treatment cytokine levels were compared with 17-OHPC serum concentrations in women with and without preterm birth. We compared serum cytokines in women with 17OHPC levels in the first quartile to women in quartiles 2-4. There was a statistically significant increase in LPS stimulated concentrations of IL-10 in women with quartiles 2-4 compared the first quartile (p<0.0001). No differences were seen prior to treatment. There was no statistically significant differences in post treatment TNF-alpha levels. After 17-OHPC administration, there appears to be a concentration relationship between 17-OHPC and LPS stimulated IL-10, an immunoinhibitory cytokine. As has been demonstrated by Harper et al 2013, IL-10 levels correlate with preterm birth. This finding supports an immunomodulatory mechanism of 17-OHPC to prevent recurrent preterm birth.View Large Image Figure ViewerDownload Hi-res image Download (PPT)

Rationale & design of the PROMISES study: a prospective assessment and validation study of salivary progesterone as a test for preterm birth in pregnant women from rural India

BackgroundIn India, 3.6 million pregnancies are affected by preterm birth annually, with many infants dying or surviving with disability. Currently, there is no simple test available for screening all women at risk of spontaneous PTB in low income setting, although high resource settings routinely use cervical length measurement and cervico-vaginal fluid fetal fibronectin for identification and care of women at risk due to clinical history. In rural India, where the public health system has limited infrastructure, trained staff and equipment, there is a greater need to develop a low-cost screening approach for providing early referral, treatment and remedial support for pregnant women at risk of preterm birth. There is interest in the use of a salivary progesterone test as a screening tool preliminary evidence from India, Egypt and UK has shown promise for this type of test. The test requires further validation in a low resource community setting.MethodsThe Promises study aims to validate and test the feasibility of introducing a low-cost salivary progesterone preterm birth prediction test in two rural districts in India with high rates of prematurity. It is a prospective study of 2000 pregnant women recruited from Panna and Satna in Madhya Pradesh over approximately 24 months. Demographic and pregnancy outcome data will be collected, and pregnancies will be dated by ultrasound sonography. Saliva progesterone will be measured by ELISA in samples obtained between 24–28 weeks of gestation. The association between salivary progesterone and preterm birth will be determined and the utility of salivary progesterone to predict preterm birth < 34, as well as < 30 and < 37 weeks assessed. Additional qualitative data will be obtained in terms of acceptability and feasibility of saliva progesterone testing and knowledge of PTB.DiscussionA validated cost-effective saliva test, which has potential for further adaptation to a ‘point of care’ setting will allow early identification of pregnant women at risk of preterm birth, who can be linked to an effective pathway of care and support to reduce preterm birth and associated adverse consequences. This will reduce both economic and emotional burden on the affected women and their families.

Cervical and Amniotic Fluid Matrix Metalloproteinase-8 and Interleukin-6 Concentrations in Preterm Pregnancies with or without Preterm Premature Rupture of Membranes

Introduction: Intra-amniotic inflammation is defined by elevated inflammatory biomarkers in the amniotic fluid (AF), either due to microbial invasion of the amniotic cavity (MIAC) or sterile inflammation. Amniocentesis being an invasive procedure, we wanted to investigate whether elevated matrix metalloproteinase-8 (MMP-8) or interleukin-6 (IL-6) concentrations could be detected from cervical fluid samples. Materials and Methods: This prospective study included 67 women with singleton nondiabetic pregnancies with or without preterm premature rupture of membranes (PPROM) between 22⁺⁰ and 37⁺⁰ weeks of gestation. Simultaneous AF and cervical samples were obtained. Results: In women without PPROM, cervical MMP-8 concentrations correlated with AF MMP-8 concentrations (r_S = 0.466, p = 0.002), but cervical IL-6 did not correlate with AF IL-6 (r_S = 0.277, p = 0.076). In PPROM cases no correlations were found. Women with MIAC had higher concentrations of AF MMP-8 and AF IL-6 compared to women without MIAC regardless of membrane status. However, only women without PPROM had higher concentrations of cervical MMP-8 in proven MIAC. Conclusion: In women without PPROM, cervical MMP-8 concentration reflects the magnitude of AF MMP-8, thus potentially guiding the selection of patients benefitting from amniocentesis.

High IL-6 and VEGF-A levels correlate with delayed wound healing in cervical lymph node tuberculosis patients

A high proportion of tuberculosis (TB) patients experience delayed wound healing after surgery, and the specific reasons for this phenomenon are not yet clear. To analyse cytokine and growth factors at surgical sites to determine their contribution to delayed wound healing in patients with cervical lymph node TB (CLNT). We measured levels of interleukin (IL) 1β, IL-6, IL-10, IL-22, epidermal growth factor, fibroblast growth factor-2, interferon-gamma, tumour necrosis factor alpha and vascular endothelial growth factor A (VEGF-A) in cervical wound drainage fluid from 36 CLNT patients who underwent lymphadenectomy and in 24 thyroidectomy patients. Wound drainage fluid from CLNT patients showed higher IL-6 (P = 0.007) and VEGF-A (P < 0.001) levels than control thyroidectomy patients (P < 0.05). CLNT patients with a delayed healing time (>5 days; n = 18) showed higher IL-6 (P = 0.002) and VEGF-A (P < 0.001) levels in wound drainage fluid than CLNT patients with normal healing times (5 days; n = 18). IL-6 (OR 11.280, 95%CI 1.413-90.028; P = 0.022) and VEGF-A (OR 13.510, 95%CI 2.168-84.182; P = 0.005) can independently and significantly predict wound healing time in CLNT patients. These findings demonstrate that high IL-6 and VEGF-A levels in the post-operative wound fluid of CLNT patients correlate with delayed wound healing.