Trends in prevalence and determinants of severe and moderate anaemia among women of reproductive age during the last 15 years in India.

BACKGROUND: Anaemia is a serious global public health problem that disproportionally affects children, adolescent girls, and women of reproductive age, especially pregnant women. Women of reproductive age are more vulnerable to anaemia, particularly severe and moderate anaemia leads to adverse outcomes among pregnant women. Despite continuous Government efforts, anaemia burden still poses a serious challenge in India. The objective of this study is to assess the trends in prevalence and determinants of severe and moderate anaemia among women of reproductive age between 15 and 49 years. METHOD: We used three rounds of the large-scale National Family Health Survey (NFHS) India, conducted on a representative sample of households using a cross-sectional design across the country in 2005-06, 2015-16 and 2019-2021. We included all the women aged 15 to 49 years in our analysis. We used the same haemoglobin (Hb) cut-off values for all the three rounds of surveys to ensure comparability. Generalized linear regression analyses with log link were done. Survey weights were incorporated in the analysis. RESULTS: The prevalence of severe or moderate Anaemia (SMA) in non-pregnant women was 14.20%, 12.43% and 13.98%; it was 31.11%, 25.98% and 26.66% for pregnant women in 2006, 2016 and 2021 respectively. The decline in SMA prevalence was 1.54% in non-pregnant women, whereas it was 14.30% in pregnant women in 15 years. Women who were poor, and without any formal education had a higher risk for severe and moderate Anaemia. CONCLUSION: Despite the intensive anaemia control program in India, SMA has not declined appreciably in non-pregnant women during the last two decades. Despite the decline, the prevalence of SMA was about 26% in pregnant women which calls for a comprehensive review of the existing anaemia control programmes and there must be targeted programmes for the most vulnerable and high-risk women such as rural, poor and illiterate women of reproductive age to reduce the burden of anaemia among them.

Utility of the lateral flow urine lipoarabinomannan tuberculosis assay in patients with advanced HIV disease at antiretroviral therapy centres in Mumbai, India.

BACKGROUND: People with Advanced HIV Disease (AHD) are at higher risk of TB coinfection and mortality. However, there are challenges in TB diagnosis with the currently recommended diagnostic tools. WHO recommends lateral flow urine lipoarabinomannan (LF-LAM) assay to assist TB diagnosis among AHD patients. We assessed the utility and acceptability of using urine LF-LAM assay for TB diagnosis among patients at public Antiretroviral Therapy (ART) Centres in Mumbai. METHODS: The cross-sectional study was conducted among adult AHD patients accessing care from 17 ART centres during November,2020-June, 2021. Urine LF-LAM was offered as routine care for eligible patients in combination with standard diagnostic tests. We calculated the proportion of positive LF-LAM results by CD4 categories and TB symptoms and performed multivariable logistic regression to determine the factors associated with LF-LAM positivity. RESULTS: Among 2,390 patients, the majority (74.5%) had CD4 between 101-200 cells/mm3. The mean age was 43.7 years (SD:10.6), 68.6% were male, 8.4% had TB symptoms and 88.0% were on ART. The overall proportion of patients with urine LF-LAM positive results was 6.4%. Among PLHIV with CD4≤100 cells/mm3, the positivity was 43.0% and 7.7% in symptomatic and asymptomatic patients, respectively. Among PLHIV with a CD4>100 cells/mm3, the positivity was 26.7% and 2.7% in symptomatic and asymptomatic patients respectively. Urine LF-LAM positivity was higher among inpatients, ART naïve, patients on treatment for <6 months, symptomatic and in WHO clinical stage III/IV of HIV disease as compared to the reference categories. We detected an additional 131 TB cases with urine LF-LAM in combination with the standard diagnostic tests. CONCLUSION: The study demonstrated the utility of urine LF-LAM for TB diagnosis among AHD patients and the simple, user-friendly test was acceptable as part of routine care. Inclusion of urine LF-LAM test in the current diagnostic algorithm may facilitate early TB diagnosis among AHD patients.

The clinical demand and supply of blood in India: A National level estimation study.

BACKGROUND: Estimating the clinical demand for blood and components arising in a health facility is crucial to ensure timely availability of blood. This study aims to estimate disease-specific clinical demand, supply and utilization of whole blood and components in India. METHODS: We conducted a national level cross-sectional study in five randomly selected states from five regions of the country. We included 251 public and private facilities representing primary, secondary and tertiary care facilities. We collected annual disease-specific demand, supply and utilization of blood and components using a structured tool. We estimated the national demand by extrapolating the study data (demand and beds) to the total number of estimated beds in the country. FINDINGS: According to the study, the total clinical demand of 251 health facilities with 51,562 beds was 474,627 whole blood units. Based on this, the clinical demand for India was estimated at 14·6 million whole blood units (95 CI: 14·59-14·62), an equivalent of 36·3 donations per 1,000 eligible populations, which will address whole blood and component requirement. The medicine specialty accounted for 6·0 million units (41·2%), followed by surgery 4·1 million (27·9%), obstetrics and gynecology 3·3 million (22·4%) and pediatrics 1·2 million (8·5%) units. The supply was 93% which is equivalent to 33·8 donations against the demand. CONCLUSION: The study indicated a demand and supply gap of 2.5 donations per 1,000 eligible persons which is around one million units. The gap emphasises the need for sustained and concerted efforts from all stakeholders and for increasing the awareness about repeat voluntary non-remunerated blood donation (VNRBD); optimizing the availability of blood components through efficient blood component separation units; promoting modern principles of patient blood management and strengthening capacities of human resources in the blood transfusion system in India.

A review of legal, regulatory, and policy aspects of blood transfusion services in India: Issues, challenges, and opportunities.

BACKGROUND: Blood transfusion services (BTS) in India have progressed significantly during the last three decades. However, there is still inequity in the availability and access to blood due to various demand and supply-side issues. Appropriate laws, regulations, policies, and guidelines are critical to ensure universal access to blood. AIMS AND OBJECTIVES: This article aims to review the evolution and current status of legal, regulatory, and policy framework and analyses the issues, challenges, and opportunities for improvement of BTS in India. METHODS: This article is based on an extensive review of currently available literature and government documents. RESULTS: The review highlights the gaps and challenges in terms of licensing, safety and quality, voluntary blood donations, the organization of BTS, access to services, and regulatory bodies. The findings emphasize the need for a coordinated response by either the National Blood Transfusion Council or a newly established autonomous "National Blood Authority" consisting of technical, administrative, and legal experts which must be exclusively responsible for regulating the BTS. As adherence to quality management systems in blood banks is not a mandatory requirement, it recommends a legal measure to ensure mandatory quality assurance in blood banks and storage centers. Towards ensuring efficiency and universal access to blood, this article recommends evidence-based criteria for establishing new blood banks to avoid skewed distribution of blood banks, component separation facilities, and blood storage centers. CONCLUSION: The review emphasizes the need for periodic reviews and updates of the legal, regulatory and policy framework, considering the rapid developments and technical advancements with increasingly complex systems and processes in transfusion medicine.

Assessment of Performance of Blood Banks in India: A National Level Cross Sectional Study.

India lacks comprehensive information about blood transfusion systems, which could help ensure universal access to safe blood through evidence-based strategies and programs. We conducted the first national assessment of blood bank systems, services, practices, and performance in India. We conducted a cross-sectional survey of all 2626 blood banks and assessed the administrative, technical, and quality aspects in 2016. In addition to descriptive analysis, we compared the means of different variables using independent t-test or ANOVA and a generalized linear model. We performed linear regression analysis between the collection of blood per 100 people and the number of blood banks per million people. The disaggregated mean score of quality management system and overall performance are presented by different groups. Besides, we graded the performance based on tertile classification, as low, medium and high-performance blood banks. Of the 2493 blood banks that participated in the study, most were public (38%) or not-for-profit(38%), and 51% had component separation facilities. Of the 11.65 million units of blood collected annually, 72% was through voluntary blood donation. There were 2.2 blood banks per one million people, collecting around one unit per 100 persons annually with wide variation between states. The mean overall performance score was 62(95% confidence interval [CI]:61.6-62.5), and the mean quality management system score was 57.4(95% CI:56.8-58.0), with significant variation across different categories of blood banks. This assessment provides critical information for developing evidence-based policies, programs, and monitoring systems to improve the performance of blood transfusion services in India.

A national level estimation of population need for blood in India.

BACKGROUND: The population need for blood is the total volume required to transfuse all the individuals who need transfusion in a defined population over a defined period. The clinical demand will arise when people with a disease or condition who require transfusion, access healthcare services, and subsequently the clinicians request blood. Essentially, the conversion of need to demand must be maximum to avoid preventable mortality and morbidity. The study estimated the population need for blood in India. METHODS: The methodology included a comprehensive literature review to determine the diseases and conditions requiring transfusion, the population at risk, and prevalence or incidence; and Delphi method to estimate the percentage of people requiring transfusion, and the quantum. RESULTS: The estimated annual population need was 26.2 million units (95% CI; 17.9-38.0) of whole blood to address the need for red cells and other components after the separation process. The need for medical conditions was 11.0 million units (95% CI:8.7-14.7), followed by surgery 6.6 million (95% CI:3.8-10.0), pediatrics 5.0 million (95% CI:3.5-7.0), and obstetrics and gynecology 3.6 million units (95% CI:1.9-6.2). The gap between need and demand which depends upon the access and efficiency of healthcare service provision was estimated at 13 million units. CONCLUSION: The study brings evidence to highlight the gap between need and demand and the importance of addressing it. It cannot be just the responsibility of blood transfusion or health systems, it requires a multi-sectoral approach to address the barriers affecting the conversion of need to clinical demand for blood.

Forecasting COVID-19 epidemic in India and high incidence states using SIR and logistic growth models.

BACKGROUND: Ever since the Coronavirus disease (COVID-19) outbreak emerged in China, there has been several attempts to predict the epidemic across the world with varying degrees of accuracy and reliability. This paper aims to carry out a short-term projection of new cases; forecast the maximum number of active cases for India and selected high-incidence states; and evaluate the impact of three weeks lock down period using different models. METHODS: We used Logistic growth curve model for short term prediction; SIR models to forecast the maximum number of active cases and peak time; and Time Interrupted Regression model to evaluate the impact of lockdown and other interventions. RESULTS: The predicted cumulative number of cases for India was 58,912 (95% CI: 57,960, 59,853) by May 08, 2020 and the observed number of cases was 59,695. The model predicts a cumulative number of 1,02,974 (95% CI: 1,01,987, 1,03,904) cases by May 22, 2020. As per SIR model, the maximum number of active cases is projected to be 57,449 on May 18, 2020. The time interrupted regression model indicates a decrease of about 149 daily new cases after the lock down period, which is statistically not significant. CONCLUSION: The Logistic growth curve model predicts accurately the short-term scenario for India and high incidence states. The prediction through SIR model may be used for planning and prepare the health systems. The study also suggests that there is no evidence to conclude that there is a positive impact of lockdown in terms of reduction in new cases.

Modelling of reproduction number for COVID-19 in India and high incidence states.

BACKGROUND: Since the onset of the COVID-19 in China, forecasting and projections of the epidemic based on epidemiological models have been in the centre stage. Researchers have used various models to predict the maximum extent of the number of cases and the time of peak. This yielded varying numbers. This paper aims to estimate the effective reproduction number (R) for COVID-19 over time using incident number of cases that are reported by the government. METHODS: Exponential Growth method to estimate basic reproduction rate R0, and Time dependent method to calculate the effective reproduction number (dynamic) were used. "R0" package in R software was used to estimate these statistics. RESULTS: The basic reproduction number (R0) for India was estimated at 1.379 (95% CI: 1.375, 1.384). This was 1.450 (1.441, 1.460) for Maharashtra, 1.444 (1.430, 1.460) for Gujarat, 1.297 (1.284, 1.310) for Delhi and 1.405 (1.389, 1.421) for Tamil Nadu. In India, the R at the first week from March 2-8, 2020 was 3.2. It remained around 2 units for three weeks, from March 9-29, 2020. After March 2020, it started declining and reached around 1.3 in the following week suggesting a stabilisation of the transmissibility rate. CONCLUSION: The study estimated a baseline R0 of 1.379 for India. It also showed that the R was getting stabilised from first week of April (with an average R of 1.29), despite the increase in March. This suggested that in due course there will be a reversal of epidemic. However, these analyses should be revised periodically.

Who is dying from COVID-19 and when? An Analysis of fatalities in Tamil Nadu, India.

BACKGROUND: As the number of COVID-19 cases continues to rise, public health efforts must focus on preventing avoidable fatalities. Understanding the demographic and clinical characteristics of deceased COVID-19 patients; and estimation of time-interval between symptom onset, hospital admission and death could inform public health interventions focusing on preventing mortality due to COVID-19. METHODS: We obtained COVID-19 death summaries from the official dashboard of the Government of Tamil Nadu, between 10th May and July 10, 2020. Of the 1783 deaths, we included 1761 cases for analysis. RESULTS: The mean age of the deceased was 62.5 years (SD: 13.7). The crude death rate was 2.44 per 100,000 population; the age-specific death rate was 22.72 among above 75 years and 0.02 among less than 14 years, and it was higher among men (3.5 vs 1.4 per 100,000 population). Around 85% reported having any one or more comorbidities; Diabetes (62%), hypertension (49.2%) and CAD (17.5%) were the commonly reported comorbidities. The median time interval between symptom onset and hospital admission was 4 days (IQR: 2, 7); admission and death was 4 days (IQR: 2, 7) with a significant difference between the type of admitting hospital. One-fourth of (24.2%) deaths occurred within a day of hospital admission. CONCLUSION: Elderly, male, people living in densely populated areas and people with underlying comorbidities die disproportionately due to COVID-19. While shorter time-interval between symptom onset and admission is essential, the relatively short time interval between admission and death is a concern and the possible reasons must be evaluated and addressed to reduce avoidable mortality.

Is BCG associated with reduced incidence of COVID-19? A meta-regression of global data from 160 countries.

BACKGROUND: Global research is running towards to find a vaccine to stop the threat of the COVID-19. The Bacillus Calmette-Guérin (BCG) vaccine that prevents severe forms of tuberculosis is getting more attention in this scenario. The objective of our study was to determine the association between BCG vaccine coverage and incidence of COVID-19 at a national-level across the Globe. METHODS: The data of 160 countries were included in the study. Meta-regression was done to estimate the difference in the incidence of COVID-19 cases between countries with BCG vaccination coverage. BCG coverage was categorized as ≤70%, >70% and no vaccination. The analyses were carried out by adjusting for factors such as population density, income group, latitude, and percentage of the total population under age groups 15-64 and above 65 years of each country. RESULTS: The countries that had ≤70% coverage of BCG vaccine reported 6.5 (95% CI: -8.4 to -4.5) less COVID-19 infections per 10,000 population as compared to countries that reported no coverage. Those that had >70% coverage reported 10.1 (95% CI: -11.4 to -8.7) less infections per 10,000 population compared to those with no BCG countries. CONCLUSION: Our analysis suggests that BCG is associated with reduced COVID-19 infections if the BCG vaccine coverage is over 70%. The region-wise analyses also suggested similar findings, except the Middle East and North African region.

Demystifying the varying case fatality rates (CFR) of COVID-19 in India: Lessons learned and future directions.

INTRODUCTION: At the end of the second week of June 2020, the SARS-CoV-2 responsible for COVID-19 infected above 7.5 million people and killed over 400,000 worldwide. Estimation of case fatality rate (CFR) and determining the associated factors are critical for developing targeted interventions. METHODOLOGY: The state-level adjusted case fatality rate (aCFR) was estimated by dividing the cumulative number of deaths on a given day by the cumulative number confirmed cases 8 days before, which is the average time-lag between diagnosis and death. We conducted fractional regression analysis to determine the predictors of aCFR. RESULTS: As of 13 June 2020, India reported 225 COVID-19 cases per million population (95% CI:224-226); 6.48 deaths per million population (95% CI:6.34-6.61) and an aCFR of 3.88% (95% CI:3.81-3.97) with wide variation between states. High proportion of urban population and population above 60 years were significantly associated with increased aCFR (p=0.08, p=0.05), whereas, high literacy rate and high proportion of women were associated with reduced aCFR (p<0.001, p=0.03). The higher number of cases per million population (p=0.001), prevalence of diabetes and hypertension (p=0.012), cardiovascular diseases (p=0.05), and any cancer (p<0.001) were significantly associated with increased aCFR. The performance of state health systems and proportion of public health expenditure were not associated with aCFR. CONCLUSIONS: Socio-demographic factors and burden of non-communicable diseases (NCDs) were found to be the predictors of aCFR. Focused strategies that would ensure early identification, testing and effective targeting of non-literate, elderly, urban population and people with comorbidities are critical to control the pandemic and fatalities.

Trends in risk behaviors among female sex workers in south India: priorities for sustaining the reversal of HIV epidemic.

HIV epidemic in India is predominantly concentrated in subgroups of population, such as female sex workers (FSWs) and their clients, whose behavior exposes them to a higher risk of acquiring HIV infection. This paper aims to present the changing patterns of socio-demographic characteristics, behaviors, reported sexually transmitted infections (STIs), and associated factors among FSWs over 11 years. Multistage cluster sampling with probability-proportional-to-size (PPS) method was used in the surveys. A sample of 400 FSWs was studied every year. The mean age and literacy at the baseline level increased significantly over the years. House-based sex increased by 40% from 43.3% in 1997 to 83% in 2008 (p<0.001). Condom use at last sex with one-time clients; consistent condom use (CCU) with one-time and regular clients indicated increasing trends. FSWs reported low levels of condom use at last sex (14.5% in 1997 to 5% in 2008; p<0.001) and CCU (12.6% in 2004 to 3.6% in 2008; p<0.01) with regular partners. FSWs who used condom with one-time clients at last sex reported significantly less STI symptoms. A two-third reduction in genital ulcers was found from 13.1% in 1997 to 4.5% in 2008 (p<0.001). Nonliterate and hotel-based sex workers were 1.6 (1.0-2.5; 95% CI) and 2.2(1.3-3.7; 95% CI) times more likely to have reported STI symptoms. The percentage of FSWs who underwent HIV testing increased (p<0.001); similarly, a 20% increase was found in FSWs who availed counseling services from 65.2% in 1997 to 85.4% in 2008 (p<0.001). Poor, illiterate, and marginalized were more likely to get involved in risky behaviors which suggest the need for structural interventions as part of HIV prevention strategy.

High tissue factor activity and low tissue factor pathway inhibitor concentrations in patients with preterm labor.

OBJECTIVE: Preterm labor (PTL) has been associated with an increased thrombin generation in the maternal circulation and amniotic fluid. Tissue factor (TF) is a potent initiator of the coagulation cascade, which can trigger the hemostatic system to generate thrombin. The aims of this study were to determine whether spontaneous PTL with intact membranes is associated with changes in the maternal plasma concentrations and activity of TF as well as tissue factor pathway inhibitor (TFPI). METHODS: This cross-sectional study included women in the following groups: (1) normal pregnancies (n = 86); (2) term pregnancies in spontaneous labor (TIL) (n = 67) and not in labor (TNL) (n = 88); and (3) patients with spontaneous PTL and intact membranes (n = 136) that were classified into three sub-groups: (a) PTL without intra-amniotic infection and/or inflammation (IAI) who delivered at term (n = 49); (b) PTL without IAI who delivered preterm (n = 54); and (c) PTL with IAI who delivered preterm (n = 33). Plasma concentrations of TF and TFPI were measured by ELISA, and their activity was measured by chromogenic assays. Non-parametric statistics were used for analysis. RESULTS: (1) Among women at term, those with spontaneous labor had a higher median maternal plasma TF and a lower median TFPI concentration than those without labor. (2) Patients with PTL had a significantly lower median maternal plasma TFPI concentration than that of normal pregnant women, regardless of the presence of IAI. (3) There was no significant difference in the median maternal plasma TF concentration between patients with a normal pregnancy and those with PTL. (4) In contrast, the median maternal plasma TF activity was higher among patients with PTL than in women with normal pregnancies, regardless of the presence of IAI or preterm delivery. (5) However, maternal plasma TFPI activity did not differ among the study groups. CONCLUSION: Women with preterm parturition, in contrast to those in labor at term, have a higher TF activity and a lower TFPI concentration, without a significant change in the median maternal plasma TF concentration. These observations suggest that the increased thrombin generation reported in patients with PTL may be the result of activation of the extrinsic pathway of the coagulation cascade. In addition, the increased thrombin generation reported in patients with PTL could be due to insufficient anti-coagulation, as reflected by the low maternal plasma TFPI concentration.

Preeclampsia and pregnancies with small-for-gestational age neonates have different profiles of complement split products.

OBJECTIVE: The activation of the complement system results in the generation of split products with pro-inflammatory properties. The objective of this study was to determine whether preeclampsia and small-for-gestational age (SGA) are associated with changes in the maternal plasma concentrations of anaphylatoxins C3a, C4a and C5a. METHODS: A cross-sectional study was conducted in the following groups: (a) normal pregnant women (n = 134); (b) women who delivered an SGA neonate (n = 53); (c) preeclampsia with (n = 52) and without SGA (n = 54). Maternal plasma anaphylatoxin concentrations were determined by enzyme-linked immunoassay. RESULTS: (1) Women with preeclampsia with or without SGA had a significantly higher median plasma C5a concentration than that of normal pregnant women and those with SGA alone (all P < 0.01); (2) women with SGA alone did not have an increase in plasma C5a concentration; (3) in contrast, the median maternal plasma concentration of C4a was lower in women with preeclampsia and SGA than that of those with a normal pregnancy (P = 0.001); (4) no changes in C3a were observed among the study groups. CONCLUSION: Preeclampsia is associated with increased plasma concentration of C5a, regardless of the presence or absence of an SGA fetus. In contrast, there was no difference in the plasma C3a, C4a and C5a concentration in patients with SGA.

Amniotic fluid sTREM-1 in normal pregnancy, spontaneous parturition at term and preterm, and intra-amniotic infection/inflammation.

OBJECTIVE: Intra-amniotic infection/inflammation (IAI) is one of the most important mechanisms of disease in preterm birth. Triggering receptor expressed on myeloid cells (TREM)- 1 is a transmembrane glycoprotein expressed by neutrophils, macrophages and mature monocytes. TREM-1 is upregulated in biological fluids and tissues infected by Gram (+) and Gram (-) bacteria and fungi, amplifies the production of pro-inflammatory cytokines and chemokines, and its soluble form (sTREM-1) is released in the presence of infection. The aim of this study was to determine the effect of gestational age, parturition (term and preterm) and IAI in the amniotic fluid (AF) concentrations of sTREM-1. STUDY DESIGN: This cross-sectional study included 434 patients in the following groups: (1) mid-trimester of pregnancy (14-18 weeks, n = 38); (2) normal pregnant women at term with (n = 39) and without (n = 39) labor; (3) patients with spontaneous preterm labor (PTL) and intact membranes classified into: (a) PTL who delivered at term (n = 99); (b) PTL who delivered preterm (<37 weeks gestation) without IAI (n = 80); and (c) PTL with IAI (n = 59); and (4) women with preterm prelabor rupture of membranes (PROM) with (n = 40) and without (n = 40) IAI. The AF concentration of sTREM-1 was determined by enzyme-linked immunoassay. Non-parametric statistics were used for analyses. RESULTS: (1) sTREM-1 was detected in all the AF samples; (2) the median AF sTREM-1 concentration at term was higher than in the mid-trimester (4277.6 pg/ml vs. 1140.4 pg/ml; p < 0.001); (3) among patients with PTL, the median AF sTREM-1 concentration was higher in patients with IAI than in those without IAI (6154.4 pg/ml vs. 3282.8 pg/ml; p < 0.001) and those with PTL who delivered at term (6154.4 pg/ml vs. 2794 pg/ml; p < 0.001); (4) patients with preterm PROM with IAI had a higher median AF sTREM-1 concentration than those without IAI (7893.1 pg/ml vs. 3386.6 pg/ml; p < 0.001); (5) no differences were observed in the median AF sTREM-1 concentration between patients with spontaneous labor at term and those at term not in labor (4712.4 pg/ml vs. 4277.6 pg/ml; respectively p = 0.4); and 6) an AF sTREM-1 concentration >or=6416 pg/ml (derived from a ROC curve) had a sensitivity of 72% and a specificity of 89% for the diagnosis of intra-amniotic infection. CONCLUSIONS: sTREM-1 is a physiologic constituent of the AF, and its concentration: (1) is significantly elevated in the presence of IAI; (2) increases with advancing gestation; and (3) does not change in the presence of spontaneous labor at term. We propose that sTREM-1 play a role in the innate immune response against intra-amniotic infection.

Amniotic fluid soluble human leukocyte antigen-G in term and preterm parturition, and intra-amniotic infection/inflammation.

OBJECTIVE: Circulating soluble human leukocyte antigen-G (sHLA-G) has been associated with pregnancy complications, and determination of sHLA-G concentrations in amniotic fluid (AF) has been reported in normal pregnancies. Our aim was to determine if the AF concentrations of sHLA-G change with advancing gestation, spontaneous labor at term, and in patients with spontaneous preterm labor (PTL) with intact membranes, as well as in those with preterm prelabor rupture of membranes (PROM), in the presence or absence of intra-amniotic infection/inflammation (IAI). STUDY DESIGN: This cross-sectional study included the following groups: (1) mid-trimester (n = 55); (2) normal pregnancy at term with (n = 50) and without (n = 50) labor; (3) spontaneous PTL with intact membranes divided into: (a) PTL who delivered at term (n = 153); (b) PTL who delivered preterm without IAI (n = 108); and (c) PTL with IAI (n = 84); and (4) preterm PROM with (n = 46) and without (n = 44) IAI. sHLA-G concentrations were determined by ELISA. Non-parametric statistics were used for analysis. RESULTS: (1) Among patients with PTL, the median AF sHLA-G concentration was higher in patients with IAI than in those without IAI or women that delivered at term (p < 0.001 for both comparisons); (2) Similarly, patients with preterm PROM and IAI had higher median AF sHLA-G concentrations than those without IAI (p = 0.004); (3) Among patients with PTL and delivery, those with histologic chorioamnionitis and/or funisitis had a higher median AF sHLA-G concentration than those without histologic inflammation (p < 0.001); and (4) The median AF sHLA-G concentration did not change with advancing gestational age. CONCLUSIONS: AF sHLA-G concentrations are elevated in preterm parturition associated to IAI as well as in histologic chorioamnionitis. We propose that sHLA-G may participate in the regulation of the host immune response against intra-amniotic infection.

Amniotic fluid fetal hemoglobin in normal pregnancies and pregnancies complicated with preterm labor or prelabor rupture of membranes.

OBJECTIVE: Hemoglobin and its catabolic products have been associated with amniotic fluid (AF) discoloration and intra-amniotic infection/inflammation (IAI). However, the origin of AF hemoglobin (maternal or fetal) has not been determined. The aims of this study were to determine if fetal hemoglobin can be detected in AF obtained from normal pregnancies, and whether there is an association between AF fetal hemoglobin concentrations and gestational age, spontaneous labor (term and preterm), preterm prelabor rupture of membranes (PPROM) and IAI. STUDY DESIGN: This cross-sectional study included pregnant women in the following groups: (1) mid-trimester (n = 60); (2) term not in labor (n = 21); (3) term in labor (n = 47); (4) spontaneous preterm labor with intact membranes (PTL) without IAI who delivered at term (n = 89); (5) PTL without IAI who delivered preterm (n = 74); (6) PTL with IAI (n = 78); (7) PPROM with (n = 48) and (8) without IAI (n = 48). AF fetal hemoglobin concentrations were determined by ELISA. Non-parametric statistics were used for analyses. RESULTS: (1) Fetal hemoglobin was detected in 80.4% of all AF samples; (2) women at term not in labor had a higher median AF fetal hemoglobin concentration than those at mid-trimester (p = 0.008); (3) labor at term was not associated with a significant difference in the median AF fetal hemoglobin concentration; (4) the median AF fetal hemoglobin concentration was not significantly different among the three PTL groups or between the PPROM groups; (5) women with PTL and IAI had a lower AF fetal hemoglobin percentage of the total hemoglobin than those without IAI who delivered preterm (p = 0.03) or at term (p < 0.001); (6) The median AF fetal hemoglobin concentration was higher in pregnancies complicated with PTL or PPROM than in women at term (p < 0.001 for all comparison). CONCLUSIONS: (1) The concentration of immunoreactive AF fetal hemoglobin increases with gestational age; (2) the median AF fetal hemoglobin concentration is higher in pregnancies complicated with PTL or PPROM than in term pregnancies; (3) among women with PTL or PPROM, the AF fetal hemoglobin concentrations were not associated with IAI; (4) however, women with PTL and IAI had a lower percentage of AF fetal hemoglobin of the total hemoglobin than those without IAI, suggesting different mechanisms of disease.

Total hemoglobin concentration in amniotic fluid is increased in intraamniotic infection/inflammation.

OBJECTIVE: Discolored amniotic fluid (AF) has been associated with intraamniotic infection/inflammation (IAI) in patients with preterm labor (PTL). The presence of hemoglobin and its catabolic products has been implicated as a cause for AF discoloration. The aim of this study was to determine whether there is an association between total hemoglobin concentration in AF and gestational age, spontaneous labor (term and preterm), and the presence or absence of IAI. STUDY DESIGN: This cross-sectional study included patients in the following groups: (1) mid trimester (n = 65 patients); (2) term not in labor (n = 22 patients); (3) term in labor (n = 47 patients); (4) spontaneous PTL who delivered at term (n = 92 patients); (5) PTL without IAI who delivered preterm (n = 76 patients); (6) PTL with IAI (n = 81 patients); (7) preterm prelabor rupture of the membranes (PPROM) with IAI (n = 48 patients); and (8) PPROM without IAI (n = 49 patients). Total hemoglobin concentrations in amniotic fluid were determined by an enzyme-linked immunoassay. Nonparametric statistics were used for analysis. RESULTS: Hemoglobin was detected in all AF samples (n = 480). The median AF total hemoglobin concentration at term was significantly higher than in mid trimester (520.6 ng/mL [interquartile range (IQR), 271.2-1549.2] vs 58.5 ng/mL [IQR, 26.1-200.8]; P < .001]). Among patients with PTL, the median AF total hemoglobin concentration was significantly higher in patients with IAI than in patients without IAI (4671.7 ng/mL [IQR, 1294.2-8620.7] vs 2013.6 ng/mL [IQR, 629.2-5420.4]; P = .01) or women who delivered at term (1143.4 ng/mL [IQR, 451.8-4037.9]; P = .001). Similarly, among patients with PPROM, the median AF total hemoglobin concentration was significantly higher in patients with IAI than in patients without IAI (10753.7 ng/mL [IQR, 2053.9-56026.6] vs 2281 ng/mL [IQR, 938.2-9191.7]; P = .02). Women at term in labor had a higher median hemoglobin concentration than did women who were not in labor (1952.6 ng/mL [IQR, 709.6-6289.2] vs 520.6 ng/mL [IQR, 271.1-1549.2]; P = .003). CONCLUSION: The AF concentration of immunoreactive total hemoglobin increases with advancing gestational age, and is elevated in pregnancies that are complicated with IAI. Spontaneous labor at term is associated with higher AF concentrations of total hemoglobin.

Microbial prevalence, diversity and abundance in amniotic fluid during preterm labor: a molecular and culture-based investigation.

BACKGROUND: Preterm delivery causes substantial neonatal mortality and morbidity. Unrecognized intra-amniotic infections caused by cultivation-resistant microbes may play a role. Molecular methods can detect, characterize and quantify microbes independently of traditional culture techniques. However, molecular studies that define the diversity and abundance of microbes invading the amniotic cavity, and evaluate their clinical significance within a causal framework, are lacking. METHODS AND FINDINGS: In parallel with culture, we used broad-range end-point and real-time PCR assays to amplify, identify and quantify ribosomal DNA (rDNA) of bacteria, fungi and archaea from amniotic fluid of 166 women in preterm labor with intact membranes. We sequenced up to 24 rRNA clones per positive specimen and assigned taxonomic designations to approximately the species level. Microbial prevalence, diversity and abundance were correlated with host inflammation and with gestational and neonatal outcomes. Study subjects who delivered at term served as controls. The combined use of molecular and culture methods revealed a greater prevalence (15% of subjects) and diversity (18 taxa) of microbes in amniotic fluid than did culture alone (9.6% of subjects; 11 taxa). The taxa detected only by PCR included a related group of fastidious bacteria, comprised of Sneathia sanguinegens, Leptotrichia amnionii and an unassigned, uncultivated, and previously-uncharacterized bacterium; one or more members of this group were detected in 25% of positive specimens. A positive PCR was associated with histologic chorioamnionitis (adjusted odds ratio [OR] 20; 95% CI, 2.4 to 172), and funisitis (adjusted OR 18; 95% CI, 3.1 to 99). The positive predictive value of PCR for preterm delivery was 100 percent. A temporal association between a positive PCR and delivery was supported by a shortened amniocentesis-to-delivery interval (adjusted hazard ratio 4.6; 95% CI, 2.2 to 9.5). A dose-response association was demonstrated between bacterial rDNA abundance and gestational age at delivery (r(2) = 0.42; P<0.002). CONCLUSIONS: The amniotic cavity of women in preterm labor harbors DNA from a greater diversity of microbes than previously suspected, including as-yet uncultivated, previously-uncharacterized taxa. The strength, temporality and gradient with which these microbial sequence types are associated with preterm delivery support a causal relationship.