Aberrant STAT3 activation and persistent expression of HPV16 E6E7 transcripts are pivotal drivers of cervical cancer (CaCx) progression. The present study was aimed to develop a Flow cytometry- based Florescence In situ hybridization (Flow-FISH) assay for simultaneous detection of STAT3 and HPV16 E6E7 transcripts at single-cell level. A set of 48 STAT3 multi locus probes and 18 HPV16 E6E7 probes were designed using Stellaris Probe Designer. Fluorescence microscopy using these probe sets generated discrete punctate signals for both individual and simultaneous hybridizations, enabling accurate transcript identification. Flow-cytometry analysis showed quantifiable STAT3 expression across CaCx cell lines, namely HeLa, SiHa and C33a. However, HPV16 E6E7 probes showed non-specific binding, which was addressed by redesigning the probes with increased stringency. The specificity of both probe sets was then evaluated through extensive sequence alignment against all known STAT3 transcript variants (n = 27) and 98 HPV16 isolates. The redesigned phase 2 HPV16 E6E7 probes were subsequently tested in cell lines, demonstrating robust detection in HPV16-positive SiHa and CaSki cells and complete absence of signal in HPV-negative controls (C33a, MSB1, SF21) or HPV18-positive HeLa cells. Dual-color flow cytometry enabled simultaneous quantification of STAT3 and HPV16 E6E7 transcripts in both cell lines and patient’s exfoliated samples. Increased dual-positive fractions across LSIL, HSIL, and SCC samples were detected that corresponded with progressive viral oncogene activity and STAT3 co-activation. Overall, the optimized probe-based Flow-FISH assay provided a sensitive, specific, and high-throughput method for transcript-level diagnostics, with potential utility for stratifying cervical lesions.

ABSTRACT:Diffuse keratinization of peritoneum can be seen in various neoplastic and non-neoplastic diseases. This exuberant keratinization exhorts a granulomatous response with multinucleated giant cell reaction. In malignancy of ovary or endometrium they mimic tumor deposits, sometimes leading to erroneous upstaging. We report one such case of ovarian endometroid adenocarcinoma in 42-year multigravida with multiple nodules over peritoneum and pelvic tissue. The case was intraoperatively staged as FIGO stage III due to nodularity over momentum and pelvic areas. Hematoxylin and eosin-stained section from the ovary revealed endometroid adenocarcinoma (FIGO grade 1) with extensive squamous morales. Sections from the omentum showed diffuse keratinization with granulomatous response against keratin. There were no viable tumor cells in the multiple sections examined. The peritoneal fluid for cytology was negative for malignancy. Final stage was given as FIGO stage IB. This case highlights this rare presentation in Ovarian endometroid adenocarcinoma posing diagnostic dilemma to clinician and pathologist and emphasizes upon the importance of clinicopathological discussion, fluid cytology and extensive sampling for accurate staging.

Cervical cancer is one of the most common gynecological malignancies worldwide. Tumor angiogenesis is a hallmark of cancer, and vascular endothelial growth factor (VEGF) is a potent angiogenic marker. Targeted therapy against VEGF in cervical carcinoma has been found to be effective and prompted us to explore other markers having a role in angiogenesis. B7-H3, an immune checkpoint molecule, is known to promote tumor progression and angiogenesis. However, the relationship between B7-H3 and angiogenesis in cervical carcinoma remains unclear. Hence, we planned to evaluate and compare the immunohistochemical expression of B7-H3 and VEGF in cervical squamous cell carcinoma. This was a retrospective study conducted on 30 histologically diagnosed cervical squamous cell carcinoma cases. Immunohistochemistry for B7-H3 and VEGF was performed in all cases. The H-score for both markers was calculated by multiplying the intensity score with the percentage cell positivity. Data was analyzed using SPSS v 20.0 software. B7-H3 and vascular endothelial growth factor immunopositivity were found in 90

To compare telomere length, hTERT expression, and telomerase activity in maternal blood of patients with spontaneous preterm birth (sPTB) and term birth (TB). One hundred and twenty participants were enrolled in this study. 60 cases of sPTB and 60 healthy controls. Whole blood was used for the estimation of Telomere Length (TL), hTERT expression, and PBMC which were used for the estimation of telomerase activity. TL and hTERT expression were measured via real-time PCR, and telomerase activity was measured using commercially available research kit. The mean (SD) TL was 239.65 (160.2) in the sPTB group and 395.18 (170.4) in the TB group (p = 0.000). There was no difference in hTERT mRNA expression and telomerase activity between sPTB and TB. sPTB is associated with shorter TL as compared to TB, even though there was no difference in telomerase activity. Therefore, shorter maternal telomere length has the potential to be a marker of sPTB. What does this study add to the clinical work: Women with inherent shorter TL are more prone to sPTB compared to healthy controls. Maternal TL could serve as a predictive marker of sPTB.

ABSTRACT Invasive stratified mucin-producing carcinoma (ISMC) is a rare, recently described subtype of HPV-associated cervical adenocarcinoma. It shows varied morphological patterns, making it difficult to diagnose, especially the mixed forms. A 57-year-old female, P7L7, presented with postmenopausal bleeding for the past five months. On per-vaginal examination, a friable cervical growth was noted. Microscopic examination of the cervical biopsy showed an invasive tumor composed of nests of stratified columnar cells with peripheral palisading and a variable amount of (alcian blue-positive) intracytoplasmic mucin. Another component of the tumor showed sheets and nests of undifferentiated tumor cells. IHC on both components showed diffuse positivity for P16, PAX8, and keratin 7. Based on these findings, a diagnosis of ISMC was made. Awareness of the morphological features and ancillary stain results of ISMC can aid in picking up the diagnosis in a small biopsy. These findings have prognostic value, as most cases of ISMC behave aggressively.

Objectives To compare vitamin A, E, and C levels in cases with idiopathic preterm premature rupture of membranes (pPROM), idiopathic spontaneous preterm birth with intact membrane (sPTB), and term birth (TB). Material and Methods There were three groups in this study: pPROM, sPTB, and TB. The sample size was 60 patients in each group (n = 60, Total = 180). Serum vitamin A and E levels and leucocyte vitamin C levels were measured using commercially available research kits. Results The mean (SD) vitamin A levels were 49.56 (18.66) µg/dL in the pPROM group, 48.67 (10.28) µg/dL in the sPTB group, and 52.69 (24.39) µg/dL in the TB group. The mean (SD) vitamin E levels were 19.17 (9.23) µg/dL in the pPROM group, 16.94 (10.17) µg/dL in the sPTB group, and 17.47 (11.19) µg/dL in the TB group. The mean (SD) vitamin C levels were 47.89 (9.53) µM in the pPROM group, 45.78 (7.92) µM in the sPTB group, and 42.35 (6.14) µM in the TB group. Vitamin C levels were significantly higher in mothers who developed pPROM (p<0.05) when compared with TB and tended toward significance in mothers who developed sPTB compared with TB. Conclusion Vitamin A and E levels were comparable in all three groups. Higher leucocyte vitamin C levels, observed in patients with pPROM (vsTB) and sPTB (vsTB), were not able to protect against pPROM and sPTB. Thus, supplementation of these vitamins during pregnancy is questionable and needs further exploration.

OBJECTIVE:We compared efficacy of weight-based (0.4 IU/kg/h) versus fixed-dose (34 IU/h) oxytocin infusion during cesarean section.METHODS:The oxytocin infusion in either group (n = 32 each) was initiated upon cord clamping. Primary outcome measure was adequacy of uterine tone at 4 min after initiating oxytocin infusion. Oxytocin associated side effects were also observed.RESULTS:Significantly less oxytocin was used with the weight-based versus fixed-dose regimen (16.3 [11.2-22.4] IU vs 20.4 [15.8-26.9] IU; P = 0.036). Incidence of adequate uterine tone was clinically greater but not significantly different with the weight-based versus fixed-dose regimen (81.3% vs 71.9%; P = 0.376). The weight-based regimen was associated with clinically lesser, although not statistically significant need for rescue oxytocin (25% vs 46.9%; P = 0.068) and additional uterotonic (9.4% vs 15.6%; P = 0.708); as well as oxytocin associated side effects (hypotension [34.4% vs 46.9%; P = 0.309], nausea/vomiting [18.8% vs 40.6%; P = 0.055], and ST-T changes [0% vs 3.1%; P = 1.000]).CONCLUSION:Weight-based oxytocin was not significantly different from the fixed-dose regimen in terms of uterotonic efficacy or associated side-effects, despite significantly lower doses being used. Use of weight-based oxytocin infusion (0.4 IU/kg/h) can be considered in clinical practice.TRIAL REGISTRATION:Clinical Trial Registry of India (ctri.nic.in, number. CTRI/2021/01/030642).

Several ion channels including calcium-activated potassium channels like KCNMA1 have been proposed as tumor markers and therapeutic targets for various cancers. KCNMA1 channel expression has been found to be progressively increasing with increasing severity of the lesion in samples collected retrospectively. Therefore, we aimed to prospectively study the mRNA and protein expression of KCNMA1 in pre-invasive and invasive cervical cancer. Sixty women were recruited to the study and allocated equally (n = 15) into four groups on the basis of histopathology, i.e., control (Group 1), cervical intraepithelial neoplasia CIN1 (Group 2), CIN2/CIN3 (Group 3) and invasive cervical carcinoma (Group 4). Real-time PCR and immunohistochemistry were used for assessing mRNA and protein expression of KCNMA1 at mRNA and protein level, respectively. The mean KCNMA1 mRNA levels in Groups 1, 2, 3 and 4 were 0.23 (SD ± 0.58), 271.40 (SD ± 1050.21), 298.84 (SD ± 1153.33) and 326.54 (SD ± 861.97), respectively (p = 0.039). Protein expression was positive in 34

To study the association of serum biomarkers (leptin and adiponectin), anthropometry and insulin resistance with endometrial cancer. This case–control study enrolled 40 women diagnosed with endometrial cancer, matched with an equal number age and BMI-matched controls. Serum leptin (ng/ml) and adiponectin (µg/ml) levels were measured using ELISA. Anthropometric measurements; waist circumference, waist/hip ratio and insulin resistance parameters, fasting insulin, and blood sugar, were assessed. Receiver operating characteristic curves were employed to evaluate the accuracy of serum biomarkers and metabolic syndrome parameters in predicting the risk of endometrial cancer, and optimal cutoff points were determined. Baseline characteristics were similar between the two groups. The mean serum adiponectin level in cases was significantly lower (7.89 ± 2.80 µg/ml versus 11.23 ± 2.84 µg/ml; p < 0.001), whereas the mean serum leptin level and Leptin/Adiponectin (L/A) ratio were significantly higher in endometrial cancer patients compared to controls (28.18 ± 13.63 ng/ml versus 12.32 ± 7.96 ng/ml, p < 0.001, and 4.02 ± 2.29 versus 1.29 ± 1.05, p < 0.001, respectively). The optimal cutoff points were identified as > 14 ng/ml for serum leptin (sensitivity: 78.9

Objective: The aim of the study was to explore the utility of fluorescein sodium (FNa) as a contrast agent for colposcopy to detect premalignant and malignant lesions of cervix. The primary objective was to determine and compare the percentage detection of premalignant and malignant lesions of FNa and acetic acid (AA) positive areas.Methods: This study included 120 screen positive women who underwent colposcopy using both 3% AA and FNa (0.06%). Observations for FNa staining were made under blue filter and directed biopsies were taken from acetowhite and fluorescent green areas. Benign lesions were considered as disease-negative and low grade squamous intraepithelial lesions (LSIL), high grade SIL (HSIL), and invasive cancer were considered as disease-positive. Correlation between histopathology and FNa and AA was determined by Kappa statistics.Results: The mean age was 39.59 +/- 10.73 years and median parity was 2. Out of 120 patients, 57 had benign lesions, 18 had LSIL, 33 had HSIL and 12 had invasive carcinomas. Sensitivity was 98.41% versus 64.91% respectively and specificity was 85.71% versus 35.09% respectively with FNa and AA. Diagnostic accuracy of FNa and AA was 82.50% versus 61.60%. There was good agreement between FNa staining and final histopathology and fair agreement between AA application and HPE (kappa = 0.643 vs 0.213, P < 0.001).Conclusion: Using FNa as a contrast agent during colposcopy results in greater accuracy for detection of premalignant and malignant lesions of the cervix as compared to AA.

The incidence of endometrial cancer is on a rise and there is paucity of definitive biomarker for screening and diagnosis of endometrial cancer which precludes early diagnosis and treatment. The primary objective of this study was to estimate and compare the levels of these biomarkers in endometrial cancer and benign endometrial pathology. The secondary objective was to correlate the biomarker levels (HE4 and fibrinogen) with various clinicopathological parameters in endometrial malignancy. A case–control study was conducted and a total of 60 patients (30 cases and 30 controls) with endometrial cancer and benign endometrial pathology, respectively, were recruited. HE4 and fibrinogen levels were estimated in both groups. The diagnostic value was assessed by a receiver operating curve, sensitivity, specificity, positive and negative predictive values and accuracy. The cutoffs calculated from the ROC curve were 239 pmol/L for HE4 and 342.5 mg/dL for fibrinogen. The mean (SD) HE4 levels were significantly higher for cases compared to controls 371.14 (258.82) pmol/L and 207.85 (179.26) pmol/L; (p = 0.017). The mean (SD) fibrinogen value for cases was 421.20 (156) mg/dl and 251 (104.4) mg/dl for controls (p = 0.00). A combination of HE4 and fibrinogen fared better than either biomarker alone in diagnosing endometrial cancer (area under the receiver operating curve: HE4 and fibrinogen = 0.86, fibrinogen = 0.81 and HE4 = 0.68). No statistically significant correlation was found between the values of these biomarkers and the pathological parameters of endometrial cancer. Both HE4 and fibrinogen were significantly raised in endometrial cancer cases than controls. Combination of serum HE4 and plasma fibrinogen had highest accuracy to differentiate benign endometrial pathology from endometrial cancer. Further prospective studies are warranted to explore their role as prognostic biomarkers.

To compare the mRNA expression of placental iron transporters (TfR-1 and FPN), markers of placental vascularization (VEGF and sFLT1) and marker of structural integrity (LMN-A) in term women with and without iron deficiency anemia. A total of 30 pregnant women were enrolled; 15 cases of iron deficiency anemia (Hb 7-10.9 gm/dL) and 15 gestational age matched healthy controls (Hb ≥ 11 gm/dL). Peripheral venous blood was collected for assessment of hemoglobin levels and serum iron profile. Placental tissue was used for assessing the mRNA expression of TfR-1, FPN, VEGF, sFLT-1 and LMN-A via real time PCR. Placental expression of TfR-1, VEGF and LMN-A was increased in pregnant women with anemia compared to healthy pregnant controls. Placental expression of sFLT-1 was decreased in pregnant women with anemia compared to healthy pregnant controls. There was no change in the placental expression of FPN. The increased expression of TfR-1, VEGF and LMN-A in cases of iron deficiency anemia are most likely to be compensatory in nature to help maintain adequate fetal iron delivery. : Compensatory changes in the placenta aimed at buffering transport of iron to the fetus are seen in pregnant women with anemia compared to healthy pregnant controls.

This study aimed to evaluate and compare the diagnostic accuracy of two ultrasound scoring systems, Assessment of Different Neoplasias in the Adnexa (ADNEX) Model and Gynecology Imaging Reporting and Data System (GI-RADS), for the preoperative assessment of adnexal masses taking histopathology as gold standard. This analytical study assessed 60 patients of age > 14 years with adnexal masses, planned for surgery. Ultrasound assessment and risk categorization according to ADNEX and GI-RADS were performed 2–3 days prior to surgery. Histopathology was used as a reference standard for the calculation of validity of two ultrasound scoring systems for diagnosis of adnexal masses. Out of 60 women (mean age, 35.52 ± 13.86 years; range, 16–70 years) with adnexal masses, 24 were malignant and 36 were benign. The sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and accuracy for the ADNEX model were 87.50

OBJECTIVE:To isolate and quantify cell-free DNA, analysis for p53 mutations, and correlation with tumor burden in women with epithelial ovarian cancer compared with benign and borderline epithelial ovarian tumors. METHODS:In this case-control study, plasma samples of eligible women collected 1 hour before surgery and based on final histopathology, women with epithelial ovarian cancer recruited as cases and borderline, and benign ovarian tumors as controls. Cell-free DNA extracted from plasma serum and quantified using Nanodrop Spectrophotometer. Amplification refractory mutation system-based polymerase chain reaction was used to detect point mutation in exon 8, codon 239 of p53 using primer pairs. p53 immunostaining was performed on tissue samples. RESULTS:A total of 40 women (20 cases of epithelial ovarian cancer and 10 each of benign and borderline ovarian tumors [controls]) were included in a 2:1:1 ratio. The mean cell-free DNA amount was 1330 ± 1705.4 ng/mL in women with epithelial ovarian cancer compared with 748.5 ± 444.8 and 448.5 ± 203.9 ng/mL in benign and borderline ovarian tumors, respectively (p = .023). In those with high-grade serous ovarian cancer, it was 2640 ± 2450.6 ng/mL compared with 600 ± 316.7 and 652.5 ± 158.9 ng/mL in low-grade serous and mucinous ovarian cancer, respectively (p = .006). In stage I and II ovarian cancer, these were 502.5 ± 134.4 and 330 ± 296.9 ng/mL, respectively, compared with 1655 ± 1924.8 ng/mL in stage III disease (p = .004). A total of 11 (55%) women with epithelial ovarian cancer harbored mutation in exon 8, codon 239 of p53 compared with 2 (20%) each in benign and borderline ovarian tumors (p = 0.07). Fair agreement was noted between cell-free DNA p53 mutation and abnormal tissue p53 staining on immunohistochemistry (κ = 0.41). CONCLUSION:Cell-free DNA amount was higher in women with epithelial ovarian cancer than women with benign and borderline ovarian tumors, with higher levels in advanced stage and high-grade serous carcinoma sub-type. Cell-free DNA p53 mutational analysis yielded fair concordance with tumor tissue p53 immunohistochemical results.

Abstract Colposcopy is the essential diagnostic tool needed to examine the cervix of women with abnormal cervical cytology or high-risk human papillomavirus and abnormal-appearing vaginal or cervical tissue. Various scoring systems have been recommended in order to improve the accuracy of colposcopic impression. Reid’s Colposcopy Index, Swede score, International Federation of Cervical Pathology and Colposcopy 2011, and most recently American Society for Colposcopy and Cervical Pathology (ASCCP) 2017 have been introduced and management recommendations for cervical screening are formulated. The new consensus ASCCP 2017 guidelines follow a risk-based approach rather than a result-based approach to determine the need for colposcopy, surveillance, and management.

Introduction/Background Lack of a definitive biomarker for screening and diagnosis of endometrial cancer precludes early diagnosis and treatment and it can only be confirmed on histopathology in symptomatic women. The aim of the present study was to examine the role of human epididymis protein 4 (HE4) and fibrinogen in endometrial cancer. The objectives were to estimate and compare the biomarker levels in endometrial cancer and benign endometrial pathology and correlate them with various clinicopathological parameters of endometrial malignancy. Methodology A total of 60 patients (30 cases and 30 controls) with endometrial cancer and benign endometrial pathology respectively were recruited in this case control study. HE4 and fibrinogen levels were estimated in both groups. The diagnostic value was assessed by a receiver operating curve, sensitivity, specificity, positive predictive value, negative predictive value and accuracy. Results The mean (SD) HE4 levels were significantly higher for cases compared to controls (371.14pmol/L (258.82) and 207.85pmol/L (179.26); (p=0.017). Similarly, the mean (SD) fibrinogen value for cases was 421.20mg/dL (156) and 251mg/dL (104.4) for controls (p=0.00).A combination of HE4 and fibrinogen fared better than either biomarker alone in diagnosing endometrial cancer (area under the receiver operating curve: HE4 and fibrinogen = 0.86, fibrinogen=0.81 and HE4=0.68). At a cut-off level of 239 pmol/L for HE4 and 342.5mg/dL for fibrinogen, the sensitivity was 60% and 73.33% respectively and specificity was 76.67% and 83.33%. No statistically significant correlation was found between the values of these biomarkers and the pathological parameters. Conclusion Both HE4 and fibrinogen were significantly raised in endometrial cancer cases than controls. Combination of serum HE4 and plasma fibrinogen had highest accuracy while plasma fibrinogen fared better as a standalone marker to differentiate between benign and malignant histology. Disclosures Nil

ObjectiveTo compare the feasibility of vagino-hysteroscopy using alginate gel Interface (VAGI) with conventional vaginoscopic hysteroscopy (CVH). MethodsThirty women undergoing diagnostic vagino-hysteroscopy were randomly allocated into Group I (VAGI): Alginate occluder was used at introitus to facilitate hydrodistension during hysteroscopy; or Group II: Underwent no-touch hysteroscopy. Primary outcome was feasibility, defined as successful visualization of uterine cavity. Secondary outcomes included operative time, hydrostatic pressures for optimum visualization, pain experienced by patient on visual analog scale, maneuverability and surgeon satisfaction. Data analysis was performed using chi(2) and Fisher exact tests for qualitative variables and Student t test for quantitative variables. ResultsVAGI was significantly better than CVH (80% vs. 33.3%; relative risk 8, P = 0.025). With VAGI, optimum visualization was achieved at significantly lower pressures at all levels (vagina, P = 0.034; cervix, P = 0.01; uterus, P < 0.001), in less time (P = 0.007), and using less irrigation fluid (P < 0.001). Surgeon satisfaction was significantly higher for VAGI (P = 0.009). Subgroup analysis showed higher likelihood of success of VAGI in women who were premenopausal (P = 0.015), younger than 45 years (P = 0.024), and had a history of vaginal birth (P = 0.03). ConclusionsVAGI is quicker to perform and provides optimum visualization at much lower pressures than CVH. Use of alginate is patient friendly and yields higher surgeon satisfaction rate.