Mukul Singh

Sustained release of microbicides by newly engineered vaginal rings.

Objective:An effective vaginal microbicide against sexual HIV transmission remains elusive, with requirements for adherence to appropriate application of effective, nontoxic products being a major deterrent. We explored methods to enable sustained release of combinations of antiretroviral microbicides, utilizing intravaginal rings composed of biosoluble Acacia gum or nonbiodegradable hydrogel of 2-hydroxyethyl methacrylate and sodium methacrylate, materials approved for use by the US Food and Drug Administration. Design and methods:The reverse transcriptase inhibitors TMC120, PMPA, 3′-azido-3′-deoxythymidine, and a newly characterized anti-HIV agent, Boc-lysinated betulonic acid, were incorporated into vaginal rings with different combinations. Daily and cumulative release rates of these inhibitors in ring eluates were determined by high-performance liquid chromatography, gas chromatography, or immunoassay. Anti-HIV effects were measured by assessment of p24 Gag antigen in T-cell cultures exposed to HIV-1 isolates. Results:Drug release rates were sustained at concentrations higher than the minimum effective dose for HIV inhibition. The release was maintained for no less than 15 and 28 days from the Acacia gum and 2-hydroxyethyl methacrylate and sodium methacrylate rings, respectively. Boc-lysinated betulonic acid showed more than 90% inhibition of HIV-1 infection in H9 cells, with little toxicity to normal cells. Conclusion:The intravaginal rings described here are capable of efficacious drug delivery. Incorporation of several antiretroviral agents, including betulinol derivatives, which act at multiple levels of the HIV life cycle, may provide a synergistic effect to achieve higher efficacy on the inhibition of HIV infection.

Investigation of the sensitivity of a cross-polarized light visualization system to detect subclinical erythema and dryness in women with vulvovaginitis

OBJECTIVE An enhanced visualization technique using polarized light (Syris v600 enhanced visualization system; Syris Scientific LLC, Gray, ME) detects surface and subsurface ( approximately 1 mm depth) inflammation. We sought to compare the Syris v600 system with unaided visual inspection and colposcopy of the female genitalia. STUDY DESIGN Erythema and dryness of the vulva, introitus, vagina, and cervix were visualized and scored by each method in patients with and without vulvitis. RESULTS Subsurface visualization was more sensitive in detecting genital erythema and dryness at all sites whether or not symptoms were present. Subsurface inflammation of the introitus, vagina, and cervix only was detected uniquely in women with vulvar vestibulitis syndrome (VVS). A subset of women presenting with VVS exhibited subclinical inflammation of the vulva vestibule and vagina (designated VVS/lichen sclerosus subgroup). CONCLUSION Enhanced visualization of the genital epithelial subsurface with cross-polarized light may assist in diagnosing subclinical inflammation in vulvar conditions heretofore characterized as sensory syndromes.

Clinical validation of enzymeimmunoassay of human luteinizing hormone (hLH) in the detection of the preovulatory luteinizing hormone (LH) surge in urine

The preovulatory luteinizing hormone (LH) surge (mean, 60.7 standard error +/- 4.7 mIU/ml) as determined by a solid-phase enzymeimmunoassay in urine has been correlated with clinical parameters in 24 women. In group A, of seven women, the preovulatory LH surge correlated with basal body temperature and cervical mucus. In one of the women in group A, serum levels of pituitary and gonadal hormones confirmed ovulation. In group B, of 17 women, the urinary estrone-3-glucuronide (E1-3-G) peak was either coincident with or preceded the LH surge. The LH surge in all cases occurred 12 to 24 hours prior to follicular rupture, as visualized by real-time sonography. The enzymeimmunoassay for the detection of the preovulatory LH surge is useful in patients for artificial insemination and for aspiration of mature oocytes for in vitro fertilization.

Verification of Early Pregnancy Tests in a Multicenter Trial

Abstract Tests for the diagnosis of early pregnancy have been available since 1974. However, no studies have systematically verified the accuracy of routine clinical laboratories in measuring human chorionic gonadotropin (hCG) prior to the time that pregnancy is clinically evident. We have conducted such a study in association with the NICHD-funded Diabetes in Early Pregnancy Study (DIEP). The purpose of this study was to elucidate the etiology of malformations in pregnancies complicated by diabetes mellitus, which probably occurs within the first few weeks of pregnancy, and therefore uniformity of pregnancy testing was necessary among the five centers to find an association of a teratogen at the time of organogenesis. We confirmed that routine clinical laboratories, in fact, could measure accurately hCG at the time of the missed menses; however, detection was not necessarily possible prior to that time. We conclude that in order to assure accurate diagnosis of early pregnancy, tests should ordinarily be delayed until time of the missed menses. When the test is used at this time, it is a reliable tool for early pregnancy testing and thus can be used to resolve questions relating to early pregnancy pathophysiology.

New approaches in fertility regulation

SummaryA Continuing search for new methods of fertility regulation has faced both promises and problems. More effective methods have led to more side effects, whereas methods with lesser problems have been less effective. Newer approaches are logically directed towards the development of contraceptive methods which are more effective and have lesser problems. Hence, newer approaches towards fertility regulation require further understanding of the phenomenon of reproduction at the cellular and molecular level and then the derivation of means to modify or intercept one or more of the earlier steps of the reproductive process. It is also vital that new approaches are (a) ethically compatible with the socio-economic, cultural, religious, and nutritional needs of a society; (b) aesthetic, preferably above the waist, and effective with little or no side effects, and (c) that they are long acting as well as of moderate cost. In view of these considerations we have directed our current approaches in the followin...

Contraceptive efficacy of norethindrone encapsulated in injectable biodegradable poly-dl-lactide-co-glycolide microspheres: phase II clinical study*

Contraceptive efficacy of injectable norethindrone (NET) microspheres of 90 day duration was evaluated for 6 months: nine women received two injections of 65 mg each and eight received two injections of 100 mg each with 90 days between each injection. Average serum NET levels were 5 to 9 ng/mL on the day after injection and subsequently remained at 1 to 3 ng/mL. With 65 and 100 mg doses, respectively, 36% and 25% of cycles had normal bleeding patterns, 36% and 48% were amenorrheic, and the remaining 28% and 27% had prolonged bleeding. Also, 56% and 40% of cycles showed increased spotting for the 65 mg and 100 mg dose, respectively. There was no incidence of pregnancy or serious side effects. The mean serum NET concentration returned to RIA nonspecific basal levels 100 days after the second injection. Women returned to ovulatory cycles between days 100 to 115 after the second injection.

Potential of LH Receptor as an Immunocontraceptive Vaccine: Studies in the Baboon (Papio)

Among the new approaches, the immunological regulation of fertility has recently received considerable attention (7,14,15). Immunization against antigens present in the ovum, sperm, and placenta (1), especially the β-subunit of the chorionic gonadotropin, has demonstrated a potential of an immunocontraceptive vaccine (15). We have examined the concept of active immunization against lutenizing hormone (LH) receptor (12) on the ovarian function of female baboons (Papio), as shown in Figure 22.1. Our observations have indicated that LH receptor has a potential for the development of a contraceptive vaccine (13).

Verification of Early Pregnancy Tests in a Multicenter Trial (42468)

Tests for the diagnosis of early pregnancy have been available since 1974. However, no studies have systematically verified the accuracy of routine clinical laboratories in measuring human chorionic gonadotropin (hCG) prior to the time that pregnancy is clinically evident. We have conducted such a study in association with the NICHD-funded Diabetes in Early Pregnancy Study (DIEP). The purpose of this study was to elucidate the etiology of malformations in pregnancies complicated by diabetes mellitus, which probably occurs within the first few weeks of pregnancy, and therefore uniformity of pregnancy testing was necessary among the five centers to find an association of a teratogen at the time of organogenesis. We confirmed that routine clinical laboratories, in fact, could measure accurately hCG at the time of the missed menses; however, detection was not necessarily possible prior to that time. We conclude that in order to assure accurate diagnosis of early pregnancy, tests should ordinarily be delayed until time of the missed menses. When the test is used at this time, it is a reliable tool for early pregnancy testing and thus can be used to resolve questions relating to early pregnancy pathophysiology.