Response of primary culture of human ovarian cancer cells to chemotherapy: In vitro individualized therapy.

OBJECTIVE: This study focused on whether primary cultures of ovarian cancer (OC) cells established from ascites can be used to evaluate response to chemotherapeutic agents and if curcumin could enhance the efficacy of these agents. MATERIALS AND METHODS: We established five primary cultures of ascitic cells from OC patients and treated them with curcumin, carboplatin, and paclitaxel singly and in combinations. The percentage of apoptotic cells was determined by flow cytometry. RESULTS: There was a wide variation in the response of individual primary cultures to treatment with the chemotherapeutic agents. Curcumin by itself was as good as carboplatin or paclitaxel in inducing apoptosis in the primary OC cells. Curcumin was not able to affect the carboplatin mediated cell killing. However, a combination of curcumin and paclitaxel was additive and was equally effective as a combination of carboplatin and paclitaxel. A combination of curcumin carboplatin, and paclitaxel was also found to be additive and, in fact, turned out to be the best combination that gave the highest percentage of apoptosis in vitro. CONCLUSION: This study highlights the fact that primary cultures of OC cells can be used to detect response to chemotherapeutic agents and help to individualize the treatment offered to OC patients.

Serum one-carbon metabolites and risk of cervical cancer.

Most cases of cervical cancer are associated with human papilloma virus (HPV) infection of high risk types. In folate deficiency, heterogeneous nuclear ribonucleoprotein E1 (hnRNP-E1) interferes with HPV16 viral capsid protein synthesis. We aimed to study the importance of 1-carbon metabolism in cervical carcinogenesis by examining serum vitamin B12 (cobalamin), homocysteine, folate levels, and the RNA and protein expression of HPV16 L1, L2, E6, E7, and to correlate them with hnRNP-E1 expression and HPV infection in normals, squamous intraepithelial lesions (SILs), and cervical cancer subjects. Serum cobalamin, folate, and homocysteine were estimated using kits, RNA by real time PCR and proteins by Western blotting. We observed that lower folate and vitamin B12 levels were associated with HPV infection. hnRNP-E1 progressively decreased from normals (100%) to SILs (75%) to cervical cancer (52.6%). The findings show that HPV16 E6 and E7 are overexpresed whereas HPV16 L1 and L2 are downregulated at mRNA and protein levels in cervical cancer as compared to normals and SILs. The results indicate that perhaps the reduced expression of hnRNP-E1 might be involved with the cervical cancer pathogenesis, with folate playing a role in the natural history of HPV infection.

A prospective randomized trial comparing the efficacy of Letrozole and Clomiphene citrate in induction of ovulation in polycystic ovarian syndrome.

OBJECTIVES: To compare the efficacy of letrozole and clomiphene citrate (CC) in patients of anovulatory polycystic ovarian syndrome (PCOS) with infertility. MATERIALS AND METHODS: This prospective randomized clinical trial included 204 patients of PCOS. 98 patients (294 cycles) received 2.5-5 mg of letrozole; 106 patients (318 cycles) received 50-100 mg of CC (both orally from Days 3-7 of menstrual cycle). The treatment continued for three cycles in both the groups. MAIN OUTCOME MEASURES: ovulation rate, endometrial thickness, and pregnancy rate. Statistical analysis was done using SPSS 13 software. P value less than 0.05 was considered significant. RESULTS: The mean number of dominant follicles in letrozole groups and CC groups was 1.86±0.26 and 1.92±0.17, respectively (P=0.126). Number of ovulatory cycle in letrozole group was 196 (66.6%) versus 216 (67.9%) in CC group (P=0.712). The mean mid-cycle endometrial thickness was 9.1±0.3 mm in letrozole group and 6.3±1.1 in CC group, which was statistically significant (P=0.014). The mean Estradiol [E2] level in clomiphene citrate group was significantly higher in CC group (364.2±71.4 pg/mL) than letrozole group (248.2± 42.2 pg/mL). 43 patients from the letrozole group (43.8%) and 28 patients from the CC group (26.4%) became pregnant. CONCLUSION: Letrozole and CC have comparable ovulation rate. The effect of letrozole showed a better endometrial response and pregnancy rate compared with CC.

Role of FDG PET-CT in detecting recurrence in patients with uterine sarcoma: comparison with conventional imaging.

PURPOSE: The purpose of the present study was to evaluate the role of 18F-fluorodeoxyglucose (FDG) PET-CT in detecting recurrent disease in posttherapy patients of uterine sarcoma and compare the same with conventional imaging (CI). METHODS: A total of 15 FDG PET-CT studies were acquired in 12 posttherapy uterine sarcoma patients. The images were evaluated by two experienced nuclear medicine physicians in consensus. Clinical/imaging follow-up (minimum 6 months) and histopathology were taken as the reference standard. All the patients had also undergone CI (CT or MRI or ultrasonography) of the chest, abdomen, and pelvis. The diagnostic accuracy of FDG PET-CT was calculated and compared with that of CI. RESULTS: The median age of the patients was 51.5 years (interquartile range: 47.5-53). Histopathology was leiomyosarcoma in six, carcinosarcoma in five, and endometrial stromal sarcoma in one patient. Six FDG PET-CT studies were carried out for suspected recurrence and nine for posttherapy surveillance. Six FDG PET-CTs were positive and nine were negative for recurrence. The sensitivity, specificity, and accuracy of FDG PET-CT were 85.7, 100, and 93.3%, respectively, on per study-based analysis, and 80, 100, and 83.3% on per lesion-based analysis. PET-CT showed higher sensitivity and specificity compared with CI for both study-based and lesion-based analysis. However, no significant difference was found between FDG PET-CT and CI either in the study-based or in the lesion-based analysis (P not significant). CONCLUSION: FDG PET-CT is a highly sensitive and specific modality for detecting recurrence in posttherapy patients with uterine sarcoma. However, it provides no significant advantage over CI for this purpose.

Postcoital vaginoperitoneal fistula after hysterectomy for gynecological malignancy.

A true incidence of postcoital fistula is not known as it is seldom suspected. It presents as acute pneumoperitoneum with signs and symptoms same as that of perforation of a hollow viscus. A 38-year-old parous woman presented with postcoital fistula 10 weeks after panhysterectomy for carcinoma ovary stage IIIc. Pneumoperitoneum was detected, with large amount of gas under the diaphragm on radiograph of the abdomen. A breach was found in the vaginal vault. All other causes of fistula were excluded. Fistula healed spontaneously on follow-up.

Human papilloma virus genotyping, variants and viral load in tumors, squamous intraepithelial lesions, and controls in a north Indian population subset.

A study of human papilloma virus (HPV) types and variants is important for developing preventive protocols and appropriate intervention targets. The presence of HPV types, their variants, and viral load in a population subset from North India was studied. Polymerase chain reaction (PCR) and line blots were used for HPV genotyping; HPV 16 and 18 viral loads were measured using real-time PCR. Variant analysis was done by sequencing of the PCR-amplified E6/E7 regions of HPV 16 and the long control region and E6/E7 regions of HPV 18. The 93.6%, 78.6%, and 10% of tumors, squamous intraepithelial lesions (SILs), and controls were HPV-positive, respectively. The most commonly observed type was HPV 16. Human papilloma virus 73 which is uncommonly observed was seen in 2 tumors. Multiple infections were more common in controls and SILs than tumors. The majority (86.4%) of the HPV 16-positive and all of the HPV 18-positive samples belonged to the European variant class. Five novel nonsynonymous changes were seen in the HPV 16-positive and 2 in HPV 18-positive samples. There was a significant increase in viral loads from controls through SILs to tumors, but no significant differences in viral loads were observed between different stages of cancer. In tumors, a significant increase in HPV 16 viral loads was seen with increasing age. The study shows a similar HPV type and variant distribution to European studies, with some differences in type distribution. Viral load does not appear to be good marker for stage wise progression and intralesional variability may affect its use as a differentiating parameter between high-grade squamous intraepithelial lesion and low-grade squamous intraepithelial lesions.

Hysterosalpingographic findings in infertile women with genital tuberculosis.

OBJECTIVES: To evaluate the hysterosalpingographic findings from infertile women who were subsequently diagnosed with genital tuberculosis. METHODS: A retrospective study of 70 infertile women who underwent hysterosalpingography to investigate infertility and were subsequently diagnosed with genital tuberculosis. RESULTS: The mean age of the women was 27.3 years and the mean duration of infertility was 6.1 years. A total of 57 (81.4%) women had primary infertility while 13 had secondary infertility. Diagnosis of genital tuberculosis was made by polymerase chain reaction (54.3%), tuberculous granuloma on biopsy (22.8%), acid-fast bacilli culture (2.8%), and at laparoscopy or hysteroscopy (20%). Hysterosalpingographic findings were a normal uterine cavity observed in 57.1% of women, an irregular cavity in 18.5%, a shrunken cavity in 2.8%, and an irregular filling defect in 18.5%. Synechiae were observed in 17.1% of women. CONCLUSION: Genital tuberculosis is a common cause of infertility in India, causing significant uterine and tubal pathologies.

Genital tuberculosis: an important cause of Asherman's syndrome in India.

OBJECTIVE: To demonstrate the association between genital endometrial tuberculosis and Asherman's syndrome. MATERIALS AND METHODS: A total of 28 women who underwent hysteroscopy with or without laparoscopy for suspected Asherman's syndrome from symptoms (amenorrhoea or oligomenorrhoea, and or primary or secondary infertility) and who were found to have genital tuberculosis on endometrial biopsy (histopathology or culture) or positive polymerase chain reaction (PCR) on endometrial aspirate or positive findings of tuberculosis on laparoscopy or hysteroscopy were enrolled in this retrospective study. RESULTS: The mean age and parity were 26.5 years and 0.3, respectively. There was past history of TB in 67.8% women. All women had menstrual dysfunction, with oligomenorrhoea and hypomenorrhoea in 16 (57%) women and amenorrhoea in 12 (42.8%). All women had primary (n = 19, 67.8%) or secondary (n = 9, 32%) infertility. On hysteroscopy, there were various grades of adhesions in all women, with grade I in 17.8%, grade II in 28.5%, grade III in 28.5% and grade IV in 17.5% women. Only four women (14.3%) had open ostia, while others had bilateral (28.5%) or unilateral (21.3%) blocked ostia or inability to see ostia (28.5%). On laparoscopy performed on 18 women, there were varying grades of adhesions in 16 (88.8%) women, with beading (33.3%), tubercles (33.3%), caseation (11.1%) and tubo-ovarian masses (11.1%). The diagnosis of genital TB was made by histopathology (tuberculous granuloma) on endometrial biopsy in 28.6%, positive culture in 3.6%, positive polymerase chain reaction (PCR) in 46.4% and observation of tubercles, beading or caseation on laparoscopy in 17.8% or shaggy cavity with caseation on hysteroscopy in 3.6% women. CONCLUSION: Genital tuberculosis appears to be an important and common cause of Asherman's syndrome in India, causing oligomenorrhoea or amenorrhoea with infertility.

Corpus luteum hemorrhage: rare complication of congenital and acquired coagulation abnormalities.

Women taking anticoagulants or those with a clotting factor deficiency are at increased risk of corpus luteum rupture due to coagulation abnormalities and three such cases are described here. Case 1 was a 35-year-old woman with prosthetic mitral valve replacement who was on anticoagulant therapy, in whom hemoperitoneum secondary to ruptured corpus luteum was seen. Emergency laparotomy revealed 1.2 L of massive hemoperitoneum. Left salpingo-oophorectomy was performed. Case 2 was two episodes of hemoperitoneum from luteal cyst rupture in a young patient with the rare congenital factor X deficiency. This patient was managed conservatively with fresh frozen plasma and blood transfusion. This is the first case of congenital factor X deficiency manifested as luteal rupture to be managed conservatively. Case 3 was two episodes of hemoperitoneum from luteal cyst rupture in a patient with antiphospholipid antibody syndrome who was on oral anticoagulants. Laparotomy was done twice with left salpingo-oophorectomy in the first instance and partial excision of the right ovary in the second instance. Hemoperitoneum secondary to rupture of the corpus luteum should be considered in the differential diagnosis of acute abdominal pain in women with congenital and acquired coagulation deficiencies.

Mutations in the mitochondrial DNA D-loop region are frequent in cervical cancer.

BACKGROUND: Mitochondrial DNA (mtDNA) is known for high mutation rates caused by lack of protective histones, inefficient DNA repair systems, and continuous exposure to mutagenic effects of oxygen radicals. Alterations in the non-coding displacement (D) loop of mitochondrial DNA are present in many cancers. It has been suggested that the extent of mitochondrial DNA mutations might be useful in the prognosis of cancer outcome and/or the response to certain therapies. In order to investigate whether a high incidence of mutations exist in mitochondrial DNA of cervical cancer patients, we examined the frequency of mutations in the D-loop region in 19 patients of cervical cancer. RESULTS: Mutations, often multiple, were detected in 18 of 19 (95%) patients. The presence of mutations correlated with Human Papilloma Virus (HPV) infection in these patients. Mutations were also detected in normal samples and lymphocytes obtained from cervical cancer patients, but their frequency of occurrence was much lower as compared to the cervical cancer tissues. CONCLUSION: Our findings indicate that D-loop alterations are frequent in cervical cancers and are possibly caused by HPV infection. There was no association of mtDNA D-loop mutations with the histopathological grade and tumor staging.