Maternal thyroid function during pregnancy as sero-diagnostic marker of pregnancy, delivery and birth outcome

1373 The Biobrio An International Quarterly Journal of Life Sciences Website: www.thebiobrio.in SJIF 2024:8.259 ISSN : 2393-9508 e-ISSN: 2582-4902 12(1&2): 1373-1386, 2025 Abhishek Mitra1, Hasib Ansari2, Birendra Kumar Gupta1, Ajay Kumar Sharma1 & Mohammad Sohail2* 1University Department of Zoology, Vinoba Bhave University, Hazaribag, Jharkhand, India 2Centre for Tribal Health and Biotechnology Foundation, New Delhi, India. Received : 28th May, 2025 ; Accepted : 28th June, 2025 DOI:- https://doi.org/10.5281/zenodo.17426336 Maternal thyroid function during pregnancy as sero-diagnostic marker of pregnancy, delivery and birth outcome ABSTRACT The burden of infection during pregnancy is of public health concern across the globe, in view of the pathogenesis, hormonal alterations, clinical sequelae, prevention and treatment of infections; all have unique features during pregnancy. The combinatorial effect of infection/s, physiological, metabolic alterations and various other obstetric complications during human pregnancy further complicates the prompt and accurate diagnosis of hormonal alterations and infections. This prompted us to investigate the prevalence of infections, haematological perturbations and the role of maternal thyroid function as reliable biochemical marker for diagnostic potential in pregnancy and delivery. Between September 2021 and December 2022, 1715 and 870 women at ANC and DU were enrolled, and screened for malaria by microscopy and RDT. Anaemia was defined as haemoglobin concentration. Thyroid function tests were measured in six stratified groups from ANC and two from DU using competitive ELISA kit. The prevalence of malaria during pregnancy was 5.4% and 4.3% at ANC and DU, respectively and 13.2% in non-pregnant women with malaria, majority of which were infected with P. vivax. Anaemia was significantly associated with malaria; however, severe anaemia was more common among women with parasitaemia. Further, observed gestational age specific increasing trend of TSH concentration and differences were significant for all the trimesters as compared to healthy women. TSH were significantly higher in infection as compared to healthy women and those with malaria without pregnancy. The TSH concentrations were highest in caesarean and still birth & higher in normal mode of delivery as compared to healthy women, whereas marginally elevated in caesarean and still birth as compared to normal delivery. Pre-term delivery had highest TSH followed by post-term and term delivery; whereas compared to term delivery, TSH was higher in pre-term and post-term delivery. High prevalence of infection in women and in pregnant women with associated anaemia suggests prompt diagnosis regardless of symptoms and comprehensive drug regime to be offered. Further, thyroid function evaluation could be considered to be reliable prognostic markers and possess promising rationale for diagnostic potential in association with existing measures in clinical spectrum of infection, pregnancy, delivery and its outcome. Key Words - TSH, Delivery, Birth-Outcome, Diagnostic Marker, Pregnancy *Corresponding author: [email protected] 1374 INTRODUCTION A number of factors make diagnosing malaria during pregnancy more difficult, such as multiple pregnancy stages with compromised immunity, heightened vulnerability to severe illnesses, different obstetric complications, parasite sequestration in the spleen and placenta, different types of anemia, and variations in patient presentation. Therefore, one of the main objectives of MIP research is the creation of an accurate and timely diagnosis. Central to this effort we evaluated the thyroid function test to identify and step towards the development of a fast, specific, safe and costeffective biochemical marker to serve as suitable and probable serodiagnostic marker. Reproductive hormones have been shown to impact thyroid physiology during pregnancy (Alexander et al., 2004 and Soldin et al., 2004) and likely lead to influence the maternal thyroid function and measures of thyroid functions (Brent, 1997). Normal maternal thyroid function during the periconception window defined as the transition from pre-pregnancy through the early first trimester is important as implantation disorders may predispose to adverse obstetrical outcomes (Lala et al., 2003) most miscarriages occur during this interval (Wang et al., 2003) and normal early fetal neurological development requires maternal thyroxin (Calyo et al., 2008). Further, maternal thyroid dysfunction during pregnancy has been shown to be associated with an increased risk of pre-term birth, placental abruption, fetal death, low birth weight infants and impaired neurological development in the child (Casey et al., 2005). Thus, in view of the prevalence, association of maternal thyroid with adverse outcomes, hormonal changes and metabolic demands occur not only in pregnancy and various trimesters, but also in labour and puerperium; prompted us to investigate with an objective to evaluate the levels of thyroid hormones in various trimesters of pregnancy and/ or with malaria and non-pregnant women with malaria as compared with healthy women to elucidate the role of thyroid regulation during malaria in pregnancy and its effect on delivery and birth outcome. Further, to explore the role of maternal thyroid function as reliable biochemical marker for diagnostic potential in malaria during pregnancy, in various modes of delivery and birthoutcome. As far as we are aware, no previous study has explored the profile, epidemiological links, and clinical correlations of malaria in pregnancy from Hazaribag, Jharkhand—one of India’s malariaendemic areas. Notably, our work represents the first attempt to examine the relationship between thyroid hormone levels and malaria during pregnancy, categorized by clinical groups in an adult population living in a region of perennial transmission where both Plasmodium vivax and Plasmodium falciparum are co-dominant (Jharkhand, Eastern India). METHODS Screening and Enrolment The study comprised three components, with recruitment focused on women attending antenatal clinics (ANC), delivery units (DU), or admitted to the antepartum ward. In the ANC group, pregnant women aged 17 years and above visiting the facility for routine care were screened, and those who provided consent were enrolled. In the DU group, women aged 18 years or older who presented for childbirth and agreed to participate through written informed consent were included. For the inpatient group, pregnant women admitted with suspected malaria, anemia, or unexplained fever were screened, and those with confirmed malaria diagnoses were enrolled after consent. At each visit, participants underwent clinical assessment, microscopic examination of peripheral blood smears, and measurement of axillary temperature prior to enrolment. The enrolment strategy, sampling procedures, and classification into broad study groups are depicted in the schematic flow chart (Figure 1). ANC procedures Trained research staff interviewed enrolled participants to obtain data on socio-demographic details (such as age, educational status, and socioeconomic background), reproductive history including gravidity, prior fever episodes, use of antiMaternal thyroid function during pregnancy as sero-diagnostic marker of pregnancy, delivery and birth outcome 1375 malarial medications, and preventive practices against malaria. A thorough clinical examination was carried out, with gestational age determined using fundal height palpation along with last menstrual period records. Axillary temperature was measured using a digital thermometer, and other vital signs were also recorded. From each participant, 3–5 ml of peripheral venous blood was DU procedures At the delivery units (DUs), enrolled pregnant women were interviewed to gather information on socio-demographic and anthropometric characteristics, obstetric complications, fever history, use of anti-malarial drugs during pregnancy, preventive practices, delivery outcomes, and mode of childbirth. Following delivery, 3–5 ml of peripheral venous blood was collected for malaria microscopy, rapid diagnostic testing (RDT), hemoglobin estimation, as well as additional biochemical and molecular analyses. Women testing positive by RDT or blood smear were referred for appropriate treatment. In addition to assessing malaria prevalence in the DU cohort, Figure-1 Schematic Flow Chart Summarizing the Sampling Strategy and Groups collected for malaria microscopy, rapid diagnostic testing (RDT), hemoglobin estimation, and additional biochemical and molecular analyses. Women who tested positive on RDT or were found to be anemic were promptly referred to the hospital physician for treatment. Furthermore, hospital staff were notified of any parasitemia detected through microscopy to ensure timely clinical management clinical and demographic information and biological samples were obtained according to mode of delivery (normal, cesarean, or stillbirth) and classified further by delivery outcomes, including pre-term, term, and post-term births. The summarized details are presented in Table 1. Table 1Baseline characteristics of pregnant women attending antenatal and delivery units Characteristics Antenatal clinics n=1271 Delivery units n=870 N, (%) N, ( %) AGE (YEARS) <20 166(13.1) 109(12.5) 20-34 983(77.4) 708(81.4) ≥35 122(9.5) 53(6.1) PRIOR PREGNANCIES Abhishek Mitra, Hasib Ansari, Birendra Kumar Gupta, Ajay Kumar Sharma & Mohammad Sohail 1376 Laboratory procedures Peripheral blood samples obtained from ANC and DU participants were used to prepare thick and thin smears, which were Giemsa-stained and examined microscopically under high power. Parasite density was estimated by counting asexual parasite stages per 200 leukocytes, assuming a standard leukocyte count of 8,000/μl of blood. Thin smears were utilized for species identification. All slides were independently verified by trained staff following strict diagnostic criteria to confirm Plasmodium infection. Additionally, the First Response Malaria pLDH/HRP2 combo rapid diagnostic test (RDT) kits (Premier Medical Corporation, Mumbai, India) were employed according to the manufacturer’s instructions as a screening tool for malaria detection in pregnant women. Hemoglobin (Hb) concentration in peripheral blood was measured using a portable HemoCue hemoglobinometer (HemoCue AB, Ängelholm, Sweden).” Sample processing and assay for thyroid function test Three to Five ml venous blood was drawn as per the sampling strategy depicted in Figure-1; before administration of antimalarial therapy, aseptically by dripping from the syringe without anticoagulant into a sterile pro-clot activator coated tubes. Blood was allowed to coagulate in a refrigerator for 4 to 6 h at 4oC before being processed by centrifugation. Sera were preserved in three to five aliquots immediately stored at -20oC, and maintained at - 80oC until measurements were performed. The TSH, T3 and T4 serum concentration were measured by competitive ELISA micro well plate-based assay using commercially available kit form Bene SpheraTM (Avantor Performance Materials, PA, USA). The assays were performed according to manufacturer’s instructions and optical densities measured using a microplate reader set to 450 nm wavelength. The sensitivity of the assay for TSH, T3 and T4 were 0.078µlU/ml, 0.04ng/ml and 0.4µg/ dl, respectively. The intra-assay TSH, T3 and T4 coefficient of variation were 5, 5.5 and 6.7%, respectively; the TSH, T3 and T4 inter-assay coefficient of variation were 6, 7 and 8.3%, PRIOR PREGNANCIES Primigravid 423(33.3) 338(38.38) Secundigravid 578(45.5) 209(24.1) Multigravid* 270(21.2) 323(37.1) GESTATIONAL AGE AT ENROLMENT (WEEKS)** <20 weeks 567(44.6) n/a 20-36 weeks 641(50.4) 57(6.5) ≥37 weeks 63(5) 813(93.5) CASTE Schedule caste 169(13.3) 93(10.7) General caste 428(33.7) 307(35.3) Other backward caste 311(24.5) 219(25.2) Scheduled trib e 363(28.5) 251(28.8) EDUCATION No formal schooling 357(28.1) 321(36.9) Attended school any length of time 914(71.9) 549(28.8) SOCIOECONOMIC CHARACTERISTICS Owns TV 567(44.6) 387(44.5) Owns bicycle 1173(92.2) 687(78.9) Owns house 958(75.4) 643(73.9) Owns refrigerator 123(9.6) 83(905) ROOF MATERIAL Mud 622(48.9) 513(58.9) Corrugated iron/asbestos sheet 242(19) 182(20.9) cement/concrete 329(25.8) 107(12.3) Other 78(6.1) 68(7.8) WALL MATERIAL Mud/sand/dung 673(52.9) 478(54.9) Mud bricks 127(9.9) 93(10.7) Cement bricks 419(32.9) 267(30.7) Other 52(4.1) 32(3.7) PRIMARY COOKING FUEL wood 619(48.7) 387(44.5) charcoal 437(34.4) 279(32.1) Gas 153(12.1) 136(15.6) Other 62(4.9) 68(7.8) MODE OF DELIVERY AMONG PREGNANT WOMEN Normal n/a 586(67.3) Caesarean n/a 179(20.6) Still Birth n/a 105(12.1) BIRTH OUTCOME Pre-Term Delivery(≤36 weeks) n/a 129(14.8) Term Delivery (31-41 weeks) n/a 623(71.6) Post-Term Delivery (after 41 weeks) n/a 118(13.5) Numbers may not add to sample size secondary to missing data. * Defined as 3 or more pregnancies ** For ANC enrollees, gestational age assessed by fundal height. For DU enrollees, gestational age was assessed by Ballard score. Maternal thyroid function during pregnancy as sero-diagnostic marker of pregnancy, delivery and birth outcome 1377 respectively. The reference interval for TSH, T3 and T4 were 0.39-6.16 µlU/ml, 0.52-1.85ng/ml and 4.411.6µg/dl, respectively. Ethics Statement and Subject Consent All blood samples in this study were obtained only after informed consent was provided by the participants, following protocols approved by the Institutional Ethics Committee (IEC) of Vinoba Bhave University, Hazaribag, Jharkhand. The procedures adhered to the ethical standards outlined by the Medical Ethics Committee, Ministry of Health, Government of India. Approval for the study protocol was granted by the IEC, VBU under memo no. VBU/R/888/2012 dated 05-06-2012. Data management and analysis All clinical, demographic, and anthropometric records were thoroughly reviewed for accuracy, and discrepancies were resolved prior to analysis. Data were entered into MS Excel and analyzed using SPSS version 16.0 (SPSS Inc., Chicago, IL, USA) and GraphPad Prism version 5.0 (GraphPad Software, CA, USA). For comparisons between two groups, Student’s t-test was applied when data followed a normal distribution, whereas the Mann–Whitney U test was used for non-normally distributed data. Categorical variables were expressed as frequencies (percentages) and compared using chisquare or Fisher’s exact test, as appropriate. Continuous variables were presented as mean ± standard error and compared using t-test or analysis of variance. Since many participants were unable to recall their exact date of birth, age was categorized into ranges based on self-reported estimates. Risk factors associated with Plasmodium falciparum or Plasmodium vivax parasitemia were first assessed by univariate analysis, followed by multivariate analysis to adjust for significant predictors. A p-value <0.05 was considered statistically significant. RESULTS Recruitment and enrolment were carried out from September 2021 to December 2022. During this period, 1,890 pregnant women attending ANC visits at Sadar Hospital, Hazaribagh, Jharkhand, were screened (Figure 2). Of these, 1,746 expressed willingness to participate, and 1,715 consented to peripheral blood sampling, while 31 declined. Thus, a total of 1,715 women were enrolled. Based on pregnancy screening, structured interviews by trained staff, and clinical assessments, participants were grouped into pregnant and non-pregnant categories. The pregnant group was further stratified by trimester, comprising 135 women in the first, 492 in the second, and 644 in the third trimester. The non-pregnant group was classified as malaria-free (227 women) and malaria-infected (217 women). At the delivery unit, 870 pregnant women were screened, all of whom were eligible and subsequently enrolled. Figure 2Map of Jharkhand Showing Study Site Antenatal clinics Most of the pregnant women attending ANC were between 18 and 38 years of age and had received some level of formal education (Table 1). A large majority were Hindi-speaking (97.6%) and nonsmokers (98.7%). About three-fourths owned their homes (75.4%) and the majority were engaged in household activities (76.7%), while a smaller proportion worked in farming (12.3%). Participants had attended a median of one ANC visit (range: 0– 9) during the current pregnancy, with nearly onethird being primigravidae (33.3%). Slightly more than half presented during the later half of pregnancy, while 44.6% reported before 20 weeks of gestation. Less than half of the women reported taking iron/ folate supplements (46.3%), and about one-third were on multivitamins (33.2%). Regarding malaria prevention, most households owned untreated bed nets and reported recent use, though very few had insecticide-treated nets (ITNs). Only nine women Abhishek Mitra, Hasib Ansari, Birendra Kumar Gupta, Ajay Kumar Sharma & Mohammad Sohail 1378 reported taking malaria prophylaxis, of whom seven (77.8%) were unable to name the medication; the remaining identified chloroquine as the drug used. Among the total cohort, malaria was confirmed in 5.4% (68/1271) of participants. Microscopic examination of blood smears identified infection in 4.3% of pregnant women, while an additional 1.1% (14 cases) were detected through rapid diagnostic tests (RDTs). Delivery units Like the ANC cohort, most pregnant women attending DUs were aged 20-36 years and had some level of formal education (Table-1). All were nonsmokers (100%) and nearly all spoke Hindi (97.2%). Most owned their own home (73.9%) and were involved in household work (84.3%); a minority engaged in farming (14.6%). Study participants had attended a median of three ANC visits (range 0-9) and about slightly less than two-thirds were primigravidae and secundigravidae (Table-1). The majority of pregnant women reported having untreated bed nets in their homes and using them recently but ITN ownership was uncommon. Only three women were taking chemoprophylaxis for malaria and none knew the name of the medication that they were taking. Only 4.3% of the women enrolled at the DUs had peripheral parasitaemia (either a positive blood smear and/or RDT). P. falciparum was identified in 5.4% (2/37), P. vivax in 86.5% (32/37), and mixed infection in 8.1% (3/37). As observed in the ANC participants, most episodes of parasitaemia occurred in July to September during the monsoon season. Nearly two third of the DU participants were anaemic whereas 16.3% had severe anaemia. For DU participants with peripheral parasitaemia, 83.7% had anaemia as compared to 47.6% of those who did not have parasitaemia (p=0.004). More women with peripheral parasitaemia had severe anaemia (5.7%) than those without parasitaemia (2.6%) and the difference was significant (p=0.02). As we observed 5.4% and 4.3% malaria during pregnancy at ANC and DU unit, respectively as compared to 1.8% and 1.7% at ANC and DU unit, respectively reported by Hamer et al. (2009) from the series of cross-sectional study in Jharkhand. However, our study design is slightly broader than the earlier investigation from Hamer et al. (2009) in terms for subject stratification, as we also taken into the account of women with malaria without pregnancy and the prevalence of malaria were found to be 13.2%, which itself reflects the importance of the investigated region and population under malaria sensitive zone. Thus, in view of the sizable prevalence, both in women with malaria without pregnancy and malaria during pregnancy, we thought to explore the serodiagnostic marker using thyroid function test. In order to investigate further, we selected 100 subjects (representative samples) each in all the groups as described previously except in women with malaria in pregnancy group (N=68) for the analysis of thyroid function tests. The experiments for this marker-based analysis were restricted with sample size due to financial constraint of the project and baseline subject characteristics are shown in Table-2. Parameters Healthy Women N=100 Mean±SE Women with Malaria N=100 Mean±SE Pregnant Women N=300 Mean±SE First Trimester N=100 Mean±SE Second Trimester N=100 Mean±SE Third Trimester N=100 Mean±SE Delivery Unit N=200 Mean±SE Pregnant Women with Malaria N=68 Mean±SE Age (years) 24.1±0.7 33.4±1.3 22.1±0.5 22.3±1.0 21.6±0.2 22.6±0.4 27.3±0.2 22.8±0.7 Weight (Kg) 49.4±1.1 47.4±0.9 47.9 ±0. 8 48.0±1.4 46.8±0.6 49.0±0.6 42 .0±0. 4 47.4±0.9 Height (Meters) 1.54±0.1 1.56±0.6 1.47±0.5 1.48±0.2 1.48±0.6 1.45±0.6 1.35±0.6 1.41±0.1 BMI (kg/m2) 20.6±0.4 19.3±0.4 22.4±0.5 22.1±0.7 22.1±0.7 23.0±0.3 22.0±0.2 24.0±0.6 Sys tolic (mm/hg) 118±2.8 119±1.8 106 ± 3 . 3 105±4.0 105 ±4.0 109±2.1 11 9±2. 3 103±2.5 Diastolic (mm/hg) 71±2.1 78±1.1 67±2.3 65±2.4 65±2.4 71±2.1 78±2.4 66±2.2 Pulse 96±2.0 96±1.0 107±3.1 110±3.9 110±3.9 101±1.2 121±1.7 105±2.3 Table 2Baseline Characteristics of the Study Population Maternal thyroid function during pregnancy as sero-diagnostic marker of pregnancy, delivery and birth outcome 1379 Thyroid Functions during Pregnancy in serum obtained from Malaria Negative Pregnant Women Little is known about thyroid function levels during normal pregnancies and to our knowledge of baseline levels at different gestational ages of Jharkhand populations have not been reported. We, therefore first determined the levels of thyroid panel (TSH, T3 and T4) in the women groups who remained infection-free and all the trimesters (from first trimester to third trimester) were observed to be significantly higher thyroid panel as compared to healthy women without pregnancy, as shown in Table-3 and Figure-3A-C. Interestingly, we observed the increasing trends of levels of the TSH are gestational age specific as compared to healthy women without pregnancy and their differences were statistically significantly for all the trimesters; P=0.005, P=0.001 and P=0.0001, respectively as shown in the Figure-3A. Further, T3 was observed to highest in second trimester and higher in first trimester as compared to third trimester, whereas T4 was highest in third trimester and higher in first trimester as compared to second trimester; although the differences were significant in all the cases as compared to healthy women as shown in Figure-3B-C. Parameters Group of Subject TSH (µIU/ml.) Mean ± SE T3 (ng/ml.) Mean ± SE T4 (µgm/ml.) Mean ± SE Healthy Women (N=100) 1.1±0.11 0.52±0.06 0.8±0.1 FirstTrimester (N=100) 1.7±0.13 0.91±0.10 1.7±0.2 Second-Trimester (N=100) 2.2±0.26 0.98±0.12 1.6±0.2 Third-Trimester (N=100) 2.4±0.24 0.81±0.07 2.1±0.3 Pregnancy with Malaria (N=68) 2.05±0.18 0.96± 0.08 1.5±0.2 Women With Malaria (N=100) 1.01±0.14 0.88±0.12 1.3±0.1 Table 3Serum triiodothyronine, thyroxin and thyroid stimulating hormone levels in the healthy women and other investigated clinical groups Figure 3 (A-C)- Circulating level of (A) TSH, (B) T3 and (C) T4 hormones among healthy women subject (HW), pregnant women in first trimester (First-TM), pregnant women in second trimester (Second-TM) and in pregnant women in third trimester (Third-TM); malaria infected pregnant women (MIP) and women with malaria (WWM). Data is presented as mean and error bar represent the plus or minus SE *p ≤ 0.01; **p ≤ 0.001; ***p ≤ 0.0001 compared with healthy subjects by two-way analysis using paired ‘t’ test through GraphPad Prism version 5.0 Abhishek Mitra, Hasib Ansari, Birendra Kumar Gupta, Ajay Kumar Sharma & Mohammad Sohail 1380 Thyroid Panel and Malaria Infection To evaluate the effect of plasmodium infection we compared the concentration of the thyroid panel (TSH, T3 and T4) in the serum of women with malaria during pregnancy and women with malaria without pregnancy as compared to healthy women as shown in Table-3 and there statistical correlation in Figure-3A-C. Most interestingly, TSH level was found to be almost double in case of malaria in pregnancy as compared to malaria-free women and malarious women with without pregnancy and differences were significant (P=0.0001, and P=0.002) (Figure-3A). Additionally, concentration of TSH was slightly lower in malaria without pregnancy as compared to healthy women and difference was not significant. Further, T3 and T4 during malaria in pregnancy were nearly double in the concentration and the differences for both were found to be significant (P=0.0001, and P=0.001, respectively) as compared to healthy women, whereas slightly higher as compared to women with malaria without pregnancy (Figure-3B-C) but in neither case (T3 and T4) did the difference reach statistical significance. Precisely, we observed that all the thyroid panel (TSH, T3 and T4) levels in malaria during pregnancy were markedly higher and their differences were statistically significant (P=0.0001, P=0.0001, and P=0.001, respectively) as compared to healthy women; whereas T3 and T4 were higher and TSH was marginally lower in case of malaria without pregnancy and their differences were statistically significant except TSH as compared to healthy women (Figure-3A-C). Assessment of Thyroid panel in Birth-Outcome and mode of Delivery To investigated the effect of thyroid function on mode and outcome of delivery, we compared the concentration of the thyroid panel (TSH, T3 and T4) in the serum of women from delivery unit; those who had delivered normal, caesarean, and still birth mode of delivery groups respectively and those who had pre-term, post-term, and term delivery groups, respectively as compared to healthy women shown in Figure-4A-C and Figure-5A-C, respectively. The TSH concentration were highest in caesarean and still birth and higher in normal mode of delivery and their differences were significant (P=0.001, P=0.01, and P=0.04, respectively) as compared to healthy women. As compared to normal delivery, we observed marginally elevated TSH in case of caesarean and still birth mode of delivery but the differences were not significant shown in Table-4. We observed highest T3 elevation in caesarean followed by normal and still birth mode of delivery and the differences were significant (P=0.001, and P=0.006, respectively) except still birth as compared to healthy women, whereas compared to normal, T3 level in caesarean and still birth mode of delivery was higher and lower, respectively and the differences were not significant. Further, T4 concentration was observed to be highest in caesarean followed by still birth and normal mode of delivery and the differences were significant (P=0.0001, P=0.006, and P=0.001, respectively), whereas compared to normal, higher levels of T4 were observed in caesarean and still birth mode of delivery although the differences were not significant. Precisely, we observed that the entire thyroid panel (TSH, T3 and T4) were reasonably elevated in caesarean and still birth mode of delivery except T3 in case of still birth as compared to normal delivery; however, the differences were not significant. We also observed that the entire thyroid panel (TSH, T3 and T4) were significantly elevated in all the mode of delivery except lower and non-significant T3 content in still birth as compared to healthy women. In addition to this, we evaluated the effect of thyroid panel (TSH, T3 and T4) test in outcome of delivery, we observed highest TSH level in pre-term delivery followed by post-term and term delivery and the differences were significant (P=0.0003, P=0.003, and P=0.04, respectively) when compared to healthy women, whereas compared to term delivery, TSH were higher in pre-term and post-term delivery and differences were significant (P=0.03, and P=0.01, respectively). Though we observed significantly elevated level of T3 in all the three Maternal thyroid function during pregnancy as sero-diagnostic marker of pregnancy, delivery and birth outcome 1381 outcome of delivery (term, pre-term and post-term delivery) compared to healthy women but the content were almost equal in all the outcome of delivery; however, the differences were significant (P=0.0001, P=0.01, and P=0.01, respectively). The T4 content was highest in pre-term delivery followed by term and post-term delivery and differences were significant (P=0.0004, P=0.0001, and P=0.0001, respectively) as compared to healthy women. Further, T4 were found to be higher in preterm and lower in post-term delivery and the differences were not significant as compared to term delivery. Precisely, we observed that the entire thyroid panel (TSH, T3 and T4) were reasonably elevated in all the three outcomes of delivery (term, pre-term and post-term delivery) and the differences were significant as compared to healthy women; whereas only TSH was found to be elevated in case of pre and post-term deliver and differences were significant as compared to term delivery. Table 4-Serum triiodothyronine, thyroxin and thyroid stimulating hormone levels in the delivery based stratification and based on the birthoutcome Parameters Groups of Subject TSH (µIU/ml.) Mean ± SE T3 (ng/ml.) Mean ± SE T4 (µgm/ml.) Mean ± SE Based on Delivery Normal Delivery 1.5±0.12 0.86±0.12 2.4±0.23 Caesarean Delivery 1.7±0.12 1.02±0.08 2.9±0.15 Still Birth 1.7±0.26 0.70±0.17 2.5±0.31 Based on Birth Outcome Pre-Term Delivery 2.1±0.24 0.93±0.17 2.8±0.30 Post-Term Delivery 1.6±0.07 0.89±0.11 2.6±0.20 Term Delivery 1.5±0.13 0.98±0.09 2.7±0.17 Figure 4 (A-C)- Circulating level of (A) TSH, (B) T3 and (C) T4 hormones based on the delivery group i.e. in normal delivery, ceserian delivery and still birth groups as compared to healthy women subject (HW). Data is presented as mean and error bar represent the plus or minus SE *p ≤ 0.01; **p ≤ 0.001; ***p ≤ 0.0001 compared with healthy subjects by two-way analysis using paired 't' test through GraphPad Prism version 5.0 Abhishek Mitra, Hasib Ansari, Birendra Kumar Gupta, Ajay Kumar Sharma & Mohammad Sohail