Use of cryopreserved cells for enabling greater flexibility in compound profiling.

Measurement of intracellular calcium release following agonist challenge within cells expressing the relevant membrane protein is a commonly used format to derive structure-activity relationship (SAR) data within a compound profiling assay. The Fluorometric Imaging Plate Reader (FLIPR) has become the gold standard for this purpose. FLIPR traditionally uses cells that are maintained in continuous culture for compound profiling of iterative chemistry campaigns. This supply dictates that assays can only be run on 4 of 5 weekdays, or alternative cell culture machinery is required such that plating can occur remotely at the weekend. The data reported here demonstrate that high-quality compound profiling data can be generated from the use of cryopreserved cells and that these cells can also be plated at various densities to generate equivalent data between 24 and 72 h post-plating. Hence, the authors report a method that allows data generation throughout the week and without the requirement of highly automated cell culture or continuous culture.

'Chemical condoms' for the prevention of HIV infection: evaluation of novel agents against SHIV(89.6PD) in vitro and in vivo.

BACKGROUND: Vaginal agents which are antiviral and/or inhibit the entry of HIV into the cell could prevent heterosexual transmission of HIV, and protect women who cannot negotiate condom use. METHODS: Four agents have been investigated for activity in vitro and in vivo against SHIV(89.6PD): two anionic polymers, dextrin-2-sulphate (D2S) and PRO 2000 (P2K), and two virucidal agents; a non-ionic detergent, nonoxynol-9 (N9) and a cyclic peptide ionophore, gramicidin-D (GD). All four agents were investigated in rhesus macaques, using an intra-vaginal challenge of two inoculations of 1 x 104 50% tissue culture infectious doses (TCID)50 of SHIV(89.6PD). RESULTS: D2S, P2K, GD and N9 all inhibited SHIV(89.6PD) in vitro. In vivo, three out of four control macaques were infected as judged by viral culture, seroconversion, DNA and RNA PCR; infection was confirmed in four out of eight macaques pre-treated with P2K, two out of four pre-treated with D2S, one out of four pre-treated with N9, two out of four pre-treated with GD and four out of four pre-treated with D2S + GD, a combination additive in vitro. INTERPRETATION: D2S and PRO-2000, novel inhibitors of HIV entry, showed evidence of protection in vivo, comparable to that seen with the virucide, N9. These data, together with the results of phase I and phase II studies in healthy women which have shown minimal toxicity, support plans for a phase III efficacy trial of chemically simple inhibitors of HIV entry with low toxicity, for the prevention of HIV infection in women.

Ethnic differences in women with HIV infection in Britain and Ireland. The study group for the mrc collaborative study of HIV infection in women.

OBJECTIVE: To examine ethnic differences in the socio-epidemiological and clinical characteristics of a cohort of women with HIV infection in Britain and Ireland. DESIGN AND METHODS: Analysis of baseline data (ethnic group, sexual history, likely route of HIV infection, reasons for HIV testing and first AIDS-defining disease) from 400 women with HIV infection recruited into a cohort study from 15 genitourinary medicine/HIV clinics in Britain and Ireland. RESULTS: Sixty-five per cent of women were white and 29% black African. Their median number of lifetime sexual partners was seven and three, respectively (P < 0.001). Ninety-three per cent of black African and 43% of white women were probably infected through sexual intercourse. Injecting drug use was the most likely route of infection in 55% of white women, but none of the black African women. Perceived risk (33%) or investigation of symptoms (26%) were the most common reasons for HIV testing. Seven per cent of white women and 16% of black African women (P < 0.001) had AIDS when HIV infection was diagnosed. The distribution of first AIDS-defining diagnoses differed (P = 0.001) by ethnic group. For white women, the most common disease was Pneumocystis carinii pneumonia; for black African women it was pulmonary tuberculosis. CONCLUSION: There are important differences between black African and white women in sexual history and route of transmission, disease stage at diagnosis and pattern of AIDS-defining diseases.

Menstrual symptoms in women infected by the human immunodeficiency virus.

OBJECTIVE: To determine whether there is a higher prevalence of menstrual symptoms in women seropositive for the human immunodeficiency virus (HIV) compared to a matched control group and to examine the relation between menstrual symptomatology and immunosuppression. METHODS: In a cross-sectional study, 55 HIV-seropositive women and a matched control group underwent detailed gynecologic assessment. The prevalence of regular cycles, oligomenorrhea, amenorrhea, menorrhagia, dysmenorrhea, and dyspareunia was assessed in the two groups. Any association with clinical disease or CD4 lymphocyte count was sought. RESULTS: There were no significant differences in the prevalence of oligomenorrhea, amenorrhea, menorrhagia, dysmenorrhea, or dyspareunia between the groups. Furthermore, no differences were demonstrated between symptomatic and asymptomatic women infected by HIV, nor was any correlation found between CD4 lymphocyte count and menstrual loss or dysmenorrhea. CONCLUSION: Infection with HIV and related immunosuppression do not seem to have a clinically significant effect on menstruation.

Rape and HIV.

The risk of contracting HIV infection as a result of rape is unknown. We describe 4 female patients who were found to have antibodies to HIV-1 following rape, only one of whom had another recognized risk factor for HIV infection. The need for careful follow-up of rape victims, and the potential for HIV acquisition by the assailant is discussed.

Changing patterns of HIV-1 transmission in southern Brazil 1985-1991.

In order to describe the changing patterns of risk factors for HIV-1 transmission of patients using hospital services at an AIDS referral centre in Porto Alegre, southern Brazil, data on demographic characteristics, referral patterns and risk factors at time of first presentation were collected prospectively on 405 patients between October 1985 and September 1991. Overall HIV-related patient workload increased during the study period, as did the proportion of infected female patients seen (P < 0.05). Of all patients, 147 (36%) presented with symptomatic HIV disease and 77 (19%) presented with an AIDS defining condition; men were more likely to present with symptomatic disease than women. Approximately 156 (44%) of men were self-referred compared with 4 (8%) of the women (P < 0.0001). Of the 357 infected men, 82 (23%) were bisexuals; of the 26 heterosexually infected women, 7 (24%) had bisexual male partners. These data suggest the increasing importance of heterosexual HIV transmission in this hitherto 'low' prevalence area, with male bisexuals constituting an important route through which heterosexual females are being infected in this area. The data also suggest that heterosexual women in Southern Brazil do not perceive themselves to be at risk for HIV-1 infection.

PIP: Unprotected homosexual intercourse and IV drug use were originally described as the predominant routes of HIV transmission in South America. Recent data, however, indicate that heterosexual transmission is becoming increasingly important in the region. With the goal of describing the changing patterns of risk factors for HIV-1 transmission, the authors collected prospectively data on demographic characteristics, referral patterns, and risk factors of patients at the time of first presentation to hospital services at an AIDS referral center in Porto Alegre, southern Brazil. The 357 male and 48 female patients presented over the period October 1985-September 1991. The overall HIV-related patient workload increased during the study period, as did the proportion of infected female patients. 44% of the men and 8% of women were self-referred. 36% of patients presented with symptomatic HIV disease and 19% with an AIDS-defining condition, with the men more likely than women to present with symptomatic disease. 23% of the infected men were bisexual; 24% of the heterosexually infected women had bisexual male partners. These findings suggest the importance of heterosexual HIV transmission in this area, with male bisexuals serving as an important route through which heterosexual females are being infected in the area. The data also suggest that heterosexual women in southern Brazil do not perceive themselves as being at risk for HIV-1 infection.

Nonoxynol-9 in lubricated condoms. Results of a study in female prostitutes.

BACKGROUND AND OBJECTIVES: Debate continues on the efficacy and safety of intravaginal nonoxynol-9 for the prevention of horizontal transmission of human immunodeficiency and other sexually transmitted diseases. Little attention has been paid to the effects of nonoxynol-9 contained in the lubricant of many condoms. GOAL: To assess the tolerability of different levels of nonoxynol-9 in condom lubricants. STUDY DESIGN: Pilot, randomized, controlled trial in 70 female prostitutes. RESULTS: There was no association between dose of nonoxynol-9 and reported symptoms or signs of genital tract inflammation; an increased dose of nonoxynol-9 was associated with increased numbers of polymorphonuclear leukocytes on a vaginal wall smear. CONCLUSIONS: There is no recognized simple method of defining inflammation in the female genital tract. Future studies of the effects of low-dose nonoxynol-9 on the female genital tract require highly controlled exposures, plus colposcopy with or without vaginal biopsy to define inflammation.

PIP: It remains unclear whether the intravaginal application of nonoxynol-9 (N-9) safely prevents the transmission of HIV and other sexually transmitted diseases (STDs) during sexual intercourse. Some studies have suggested a protective effect, while others a potential adverse effect created through the inflammation and ulceration of the genital tract caused by N-9. N-9 is commonly found in the lubricating agents used on condoms. However, the dose in each condom is small relative to pessaries or spermicidal gels. 70 female prostitutes participated in a study to assess the tolerability of different levels of N-9 in condom lubricants. Women were excluded from participation if they had a STD, vaginal candidiasis or bacterial vaginosis, were pregnant, or were using intravaginal spermicides for contraception. Participants were asked to use the trial condoms for all vaginal intercourse for a 2-week period and to avoid using any other spermicidal preparation. 1636 identical condoms were used during the study, either lubricated with polyethyleneglycol, polyethyleneglycol with 2% N-9, or polyethyleneglycol with 4% N-9. No association was found between the dose of N-9 and reported symptoms or signs of genital tract inflammation, although an increased dose of N-9 was associated with increased numbers of polymorphonuclear leukocytes on a vaginal wall smear. Future studies of the effects of low-dose N-9 on the female genital tract require highly controlled exposures together with colposcopy with or without vaginal biopsy for confirmation.

Gummatous penile ulceration and generalised lymphadenopathy in homosexual man: case report.

A case of granulomatous penile ulceration associated with generalised lymphadenopathy in a homosexual man aged 43 is presented. His serum gave positive results to tests for syphilis, but was negative for antibody to human immunodeficiency virus (HIV). His condition responded briskly to treatment with procaine penicillin, and we conclude that the clinical features were attributable to infection with Treponema pallidum.

HIV infection and the gynaecologist.

PIP: As women infected with human immunodeficiency virus (HIV) will soon account for 1 in 500 gynecological patients in Great Britain, gynecologists have an obligation to become informed about the transmission, clinical manifestations, and management of HIV. Women with HIV infection are at increased risk of lower genital tract neoplasia with extensive cervical, vaginal, and vulvar lesions. There is also a strong association between HIV infection and sexually transmitted diseases involving genital ulcerations. In fact, herpes, warts, and candidiasis may be the initial clinical presentation of HIV infection. Estradiol levels may fall in seropositive women, but, in general, menstruation and ovulation are maintained. Latex condoms remain the only contraceptive choice for HIV-infected women. Sufficient data have not been accumulated on the effectiveness of the female condom, but it has the potential of giving women in developing countries in particular greater control over HIV prevention. Condom use is essential even if both partners are HIV-positive since the acquisition of different HIV strains can accelerate disease progression. Given their higher risk of percutaneous in injury compared to other surgeons, gynecologists should use double gloving and blunt tipped needles and staples.^ieng

Is endometrial infection with Candida albicans a cause of recurrent vaginal thrush?

OBJECTIVE: It was hypothesised that the endometrium might act as a reservoir for candida, thus infecting the vagina as the endometrium is shed during menstruation. DESIGN: A prospective study of women with recurrent vulvo-vaginal candidiasis. The endometrium was sampled and cultured for candida species. SETTING: Central London STD clinic. SUBJECTS: 26 women were enrolled, of whom 20 completed the study. RESULTS: One patient had a positive endometrial culture for candida species, the isolate being Candida krusei. CONCLUSIONS: The endometrium is not a common resevoir for candida species and therefore, infection at this site is an unlikely cause of recurrent vaginal candidiasis.

Immunity in the female lower genital tract and the impact of HIV infection.

This study investigates the distribution of immunocompetent cells in the ectocervix, and cytokine and immunoglobulin (Ig) levels in cervicovaginal secretions to determine whether they are altered in asymptomatic human immunodeficiency virus (HIV) infection. Ectocervical biopsies from 10 HIV+ and 10 presumed HIV-ve women were studied by immunocytochemistry. Levels of Igs in cervicovaginal secretions were quantified by radial immunodiffusion (RID) and cytokine levels by ELISA. HIV+ women had significantly increased numbers of CD8+ lymphocytes resulting in reversal of the CD4:CD8 ratio. There was a significant increase in the proportion of activated CD8+ HLA-DR+ and CD4+ HLA-DR + lymphocytes, but not in CD8+ TIA-1+ cells. The epithelium of the cervix from HIV+ subjects showed a significant increase in both numbers of macrophages (CD68+) and proportions of activated macrophages (CD68+ HLA-DR+) compared to normal. The stroma contained increased proportions of inductive (D1+) and suppressive (D1+ D7+) macrophages but a decrease in effector phagocyte (D7+) proportions and Langerhans' cells. Significantly lower tumour necrosis factor (TNF)-alpha levels were observed in cervicovaginal secretions from HIV+ subjects. IgG levels were 4 times higher and IgM levels twice higher in cervicovaginal secretions from HIV+ women, compared to results from normal subjects. These results suggest a response within the CD8+ cells in HIV+ women, yet these cells may have a low cytolytic capacity. The raised proportions of HLA-DR+ and D1+ CD4+ macrophages could act as antigen-presenting cells (APC) for CD4+ CD45RO+ lymphocytes, and represent a local acquired response. However, the close juxtaposition of these cells offers the potential for them to act as a local reservoir of virus and promote its proliferation. The increase of IgG over sIgA in secretions of HIV+ subjects provides evidence suggesting a dysregulation of local humoral immunity.

Gummatous lesions in men infected with human immunodeficiency virus and syphilis.

Two HIV-infected men are reported who developed gummatous lesions more than 12 months after appropriate treatment of presumptive syphilis. In one patient the lesions developed without any change in the VDRL titre. The most likely explanation for these lesions is reactivation of syphilis in the context of HIV infection. As these lesions respond to penicillin, the possible reactivation of appropriately treated syphilis, or even yaws, should now be considered in any ulcerative lesion in HIV infected individuals at risk from treponemal infection.

Comparison of ofloxacin with doxycycline in the treatment of non-gonococcal urethritis and cervical chlamydial infection.

The efficacy of ofloxacin was compared with that of doxycycline in the treatment of non-gonococcal urethritis (NGU) and cervical chlamydial infection in an open randomized study. In total, 182 men and 100 women were enrolled: 92 men and 50 women received ofloxacin at a dose of 400 mg once daily for seven days, and 90 men and 50 women received doxycycline at a dose of 100 mg twice daily for seven days. All women and 42 men, that is 29 men in the ofloxacin group (32%) and 13 in the doxycycline group (14%) were chlamydia-positive on entry into the study. Patients were reviewed on completion of the treatment course (day 8) and two weeks later (day 22). Clinical cure rates for NGU in assessable patients in each treatment group were similar; 73% in the ofloxacin group and 63% in the doxycycline group at day 8, and 70% and 90% in the respective treatment groups on study completion. Repeat tests for chlamydiae were negative for all male and female patients at both day 8 and day 22. Both ofloxacin and doxycycline were well tolerated. We conclude that ofloxacin given at a dose of 400 mg once daily for seven days is a safe and effective alternative treatment for NGU and cervical chlamydial infection.