In vitro and In vivo evidence demonstrating chronic absence of Ref-1 Cysteine 65 impacts Ref-1 folding configuration, redox signaling, proliferation and metastasis in pancreatic cancer.

Ref-1/APE1 (Redox Effector/Apurinic Endonuclease 1) is a multifunctional enzyme that serves as a redox factor for several transcription factors (TFs), e.g., NF-kB, HIF-1α, which in an oxidized state fail to bind DNA. Conversion of these TFs to a reduced state serves to regulate various biological responses such as cell growth, inflammation, and cellular metabolism. The redox activity involves a thiol exchange reaction for which Cys65 (C65) serves as the nucleophile. Using CRISPR editing in human pancreatic ductal adenocarcinoma (PDAC) cells, we changed C65 to Ala (C65A) in Ref-1 to evaluate alteration of Ref-1 redox dynamics as well as chronic loss of Ref-1 redox activity on cell signaling pathways, specifically those regulated by NF-kB and HIF-1α. The redox activity of Ref-1 requires partial unfolding to expose C65, which is buried in the folded structure. Labeling of Ref-1 with polyethylene glycol-maleimide (PEGm) provides a readout of reduced Cys residues in Ref-1 and thereby an assessment of partial unfolding in Ref-1. In comparing Ref-1WT vs Ref-1C65A cell lines, we found an altered distribution of oxidized versus reduced states of Ref-1. Accordingly, activation of NF-kB and HIF-1α in Ref-1C65A lines was significantly lower compared to Ref-1WT lines. The bioinformatic data revealed significant downregulation of metabolic pathways including OXPHOS in Ref-1C65A expressing clones compared to Ref-1WT line. Ref-1C65A also demonstrated reduced cell proliferation and use of tricarboxylic acid (TCA) substrates compared to Ref-1WT lines. A subcutaneous as well as PDAC orthotopic in vivo model demonstrated a significant reduction in tumor size, weight, and growth in the Ref-1C65A lines compared to the Ref-1WT lines. Moreover, mice implanted with Ref-1C65A redox deficient cells demonstrate significantly reduced metastatic burden to liver and lung compared to mice implanted with Ref-1 redox proficient cells. These results from the current study provide direct evidence that the chronic absence of Cys65 in Ref-1 results in redox inactivity of the protein in human PDAC cells, and subsequent biological results confirm a critical involvement of Ref-1 redox signaling and tumorigenic phenotype.

Pregnancies after reconstructive surgery for congenital cervicovaginal atresia.

Eighteen women aged 12 to 20 years with congenital cervicovaginal atresia were treated with a new technique of surgical canalization. All are now having regular menses. Two pregnancies have been achieved, with delivery of viable neonates. Surgical correction of this anomaly during adolescence may aid in restoration of menstrual and reproductive function.

Overview of tuberculosis of the female genital tract.

PIP: Female genital tuberculosis is not uncommon in countries where pulmonary tuberculosis is widespread. Improved diagnostic techniques (e.g., endometrial curettage and biopsy, histologic examination of curettage, hysterosalpingography, bacteriologic examination of menstrual blood, laparoscopy, and ultrasonography) have resulted in increased awareness of genital tuberculosis. Another trend has been heightened awareness of the potential role of female genital tuberculosis in infertility; this infection has been implicated in 5-10% of infertility cases. Genital organs most frequently affected include fallopian tubes (95-100%), endometrium (50-60%), and ovaries (20-30%). In 92% of cases, genital tuberculosis is secondary to a focus in the lungs, lymph nodes, urinary tract, bones, or joints. Given the hormone-dependent nature of female genital tuberculosis, 90% of cases involve women under 40 years of age. The major presenting symptoms are infertility (45-55%), pelvic pain (50%), poor general health (25%), and menstrual disturbances (20%). The differential diagnosis includes chronic pelvic inflammation, mycotic infection, enterobiasis, lipid salpingitis, and carcinoma. Female genital tuberculosis is treated with the same long-term, combined drug therapy used in pulmonary and extrapulmonary tuberculosis. Surgery should be undertaken only after continuous drug treatment of 12-18 months' duration. In women of childbearing age, an attempt can be made to preserve one ovary. Successful pregnancy is unlikely, however, after complete antituberculosis treatment or tuboplastic surgery.^ieng

Problems of AIDS in India especially in women.

According to the World Health Organisation survey, 3 million women are already infected with HIV all over the World, but in India the problem of AIDS as a whole is not that acute especially in women. At least it is not that alarming. India, having a very high incidence of STD, it will not take much time for the spread of the HIV infection. No case of mother-to-infant transmission of HIV and resultant clinical disease in the neonate and children has as yet been reported in India. Infected babies, however, face a short life--nearly all die before reaching 2 years of age. Breast feeding is not a significant means of transmitting AIDS. The disease can spread only through sexual contact, use of infected syringe or transfusion of infected blood. Therefore, doctors, nurses and other paramedical staff in hospitals who may have to deal with AIDS patients need not fear the virus if they take adequate precautions. There is no known curative treatment for HIV infection causing AIDS, but improving literacy, avoiding sexual promiscuity and using condoms in heterosexual intercourse are positive steps in keeping HIV infection at a considerably low level.

PIP: Although data from a World Health Organization survey indicate that 3 million women are infected with HIV worldwide, AIDS in India in general and especially among women has yet to become an acute problem. One should take note, however, that 60% of HIV infection over the world is the result of heterosexual intercourse. This proportion will increase to 75-80% by the year 2000, making women in their subordinate familial and social roles even more vulnerable to HIV infection from sexual intercourse. The very high incidence of sexually transmitted diseases in India will help facilitate the rapid spread of HIV. Accordingly, unless urgent steps are taken to stem the spread of HIV, HIV and AIDS will sweep over India in massive, destructive proportions in the coming years. Although no case of maternal-infant HIV transmission and resultant clinical disease in neonates and children has been reported, the risk of maternal-fetal and maternal-infant infection remains. Breastfeeding is not considered a significant means of transmitting AIDS. Insofar as the safety from infection of health professionals who care for people with AIDS is concerned, professionals must remember that HIV is transmitted through sexual contact, use of infected syringes, or the transfusion of infected blood. Health professionals, therefore, need not fear being infected by their patients as long as they adhere to proper precautionary procedures. Finally, while no curative treatment is known for HIV infections, improving literacy, avoiding sexual promiscuity, and using condoms in sexual intercourse will help minimize the incidence of HIV infection.

Risk of carcinogenesis among pill users.

PIP: Recently, Bishop and Wilson reviewed the evidence regarding a relationship between estrogen administration and cervical cancer. They separately concluded that there is no evidence that administered estrogens have a carcinogenic effect. It appears tht the balance of action of oral contraceptive (OC) tablets is progestogenic and progestogens has a protective effect in humans, although experimental work on laboratory animals has tended to confirm suspicions about the carcinogenicity of administered estrogen. From an epidemiological perspective, there is little reason to suppose that steroidal contraceptives would have any influence on cervical cancer. Several studies of the relationship between OCs and carcinoma of the cervix in situ have been undertaken. A study conducted by Melamed et al. included women who had used OCs for 1 year or more and diaphragm users who were attending family planning clinics in New York City. The subjects were individually matched taking into account 5 variables -- age, parity, age at 1st pregnancy, ethnic group and family income. The prevalence of carcinoma in situ was found to be about twice as high among those who used OCs than among diaphragm users. The result may be interpreted in at least 3 ways: the OCs caused the development of carcinoma in situ; the diaphragm protected against it; or the women who chose OCs were more likely, for reasons not considered in the matching procedure, to develop the disease. The studies of Thomas and Engineer et al. are also reviewed.^ieng

Sociological implications of pregnancy in unmarried teen-agers.

PIP: A critical analysis was made in 66 pregnancies in random unmarried teen-agers attending the Medical Termination of Pregnancy (MTP) Clinic, Eden Hospital, Calcutta, India over the 1976-1977 period in order to examine some of the sociological problems associated with unwanted pregnancies. The annual number of adolescent out-of-wedlock pregnancies has increased steadily in recent years, with the adolescent increase proportionately somewhat greater than the overall increase. The increase in sexual promiscuity must have some relation with liberalization of abortion and easy availability of the abortion service since the introduction of the Medical Termination of Pregnancy Act. Although teen-agers pregnant out-of-wedlock came from all segments of society, the data collected showed that more came from the lower socioeconomic group, low educational standard, poor residential status and with very little knowledge of reproductive biology and contraceptive practice. The teen-age females who became pregnant in this series were grossly uninformed about the human body and sexual relations. These young women tried to conceal their pregnancies, mostly in order to keep from confronting their parents. As a result, most of the cases sought abortion services between 16-20 weeks (47 out of 66). Along with enlightened sex education programs, it is necessary to provide fertility regulation to adolescents in properly selected cases and in a congenial atmosphere.^ieng