Modulation of TNFalpha, a determinant of acute toxicity associated with systemic delivery of first-generation and helper-dependent adenoviral vectors.

Understanding the determinants of the host innate immune response to systemic administration of adenoviral (Ad) vectors is critical for clinical gene therapy. Acute toxicity occurs within minutes to hours after vector administration and is characterized by activation of innate immune responses. Our data indicate that in mice, indicators of vector toxicity include elevations of cytokine levels, liver transaminase levels and thrombocytopenia. To discern potential targets for blunting this host response, we evaluated genetic factors in the host response to systemically administered first-generation Ad vectors (FGV) and helper-dependent Ad vectors (HDV) containing beta-galactosidase expression cassettes. A preliminary screen for modulation of vector-induced thrombocytopenia revealed no role for interferon-gamma, mast cells or perforin. However, vector-induced thrombocytopenia and interleukin 6 (IL-6) expression are less evident in tumor necrosis factor alpha (TNFalpha)-deficient mice. Moreover, we also demonstrated that TNFalpha blockade via antibody or huTNFR:Fc pretreatment attenuates both thrombocytopenia (>40% increase in platelet count) and IL-6 expression (>80% reduction) without affecting interleukin 12 , liver enzymes, hematological indices or vector transduction in a murine model. Our data indicate that the use of HDV, in combination with clinically approved TNFalpha immunomodulation, may represent an approach for improving the therapeutic index of Ad gene therapy for human clinical trials.

Helper-dependent adenoviral gene therapy mediates long-term correction of the clotting defect in the canine hemophilia A model.

BACKGROUND: Adenoviral vector-mediated gene therapy might have potential for long-term correction of the monogenic disease hemophilia A. OBJECTIVE: In this study, we tested the efficacy of administering a helper-dependent adenoviral vector (HDV) designed for maximal liver-restricted canine factor VIII (cFVIII) expression on three out-bred hemophilia A dogs. METHODS: Three FVIII-deficient animals from the University of North Carolina colony were injected with 1 x 10(12) (Dog A), and 3 x 10(12) (Dog B and C) vp kg(-1) helper-dependent adenoviral vector, and we performed systematic analysis of toxicity, persistence of therapeutic gene expression, and molecular analysis of gene transfer. RESULTS: We observed acute dose-dependent elevation in liver enzymes and thrombocytopenia after injection, although both were transient and resolved within 2 weeks. The whole blood clotting time (WBCT), plasma FVIII concentration, FVIII activity, and activated partial thromboplastin time in all animals improved significantly after treatment, and two animals receiving a higher dose reached near normal WBCT with low-level FVIII activity until terminal sacrifice at 3 months, and 2 years. Importantly, the treated dogs suffered no bleeding events after injection. Moreover, we observed persistent vector-specific DNA and RNA in liver tissue collected from one high-dose animal at days 18 and 79, and could not detect the formation of inhibitory antibodies. CONCLUSION: Although vector-associated toxicity remains an obstacle, a single injection of HDV led to long-term transgene expression and vector persistence in two FVIII-deficient animals with conversion of their severe phenotype to a moderate one.

Epidemiology and natural history of ligase chain reaction detected chlamydial and gonococcal infections.

OBJECTIVES: Ligase chain reaction (LCR) technology has dramatically increased the sensitivity of tests for sexually transmitted infections (STIs). It is unknown whether low copy infections (LCR positive, culture negative) have any clinical consequences. We assessed the clinical significance of untreated low copy Chlamydia trachomatis and Neisseria gonorrhoeae infections in a cohort of sexually active women. METHODS: We studied a cohort of sexually active women followed at 6 month intervals for up to 3 years. Frozen urine specimens from 181 women with negative cultures for C. trachomatis and N. gonorrhoeae who were 'high risk' (defined as being less than 40 years old at baseline, and having either Trichomonas vaginalis at baseline or a history of more than one sexual partner during the 12 months before baseline) were tested for C. trachomatis and N. gonorrhoeae by LCR (Abbott Laboratories, Abbott Park, IL, USA). The specimens from all visits for each person were pooled and LCR was performed on the pool. Laboratory results were linked to clinical information. We also tested all urine samples obtained from patients with a positive culture. RESULTS: 10 additional infections (nine C. trachomatis and one N. gonorrhoeae) were detected with LCR technique. None of the women with low copy infection had evidence of subsequent pelvic inflammatory disease or ectopic pregnancy. Pooling of urine samples resulted in a 47% decline in the number of tests performed. CONCLUSIONS: Additional STIs can be identified when using LCR. Pooling of urine specimens is a cost saving technique for C. trachomatis and N. gonorrhoeae testing.

Comparison of clindamycin phosphate vaginal cream with triple sulfonamide vaginal cream in the treatment of bacterial vaginosis.

BACKGROUND: Triple sulfonamide vaginal cream has been used to treat bacterial vaginosis for many years. There are few studies in which triple sulfonamide cream has been compared with newer regimens. GOAL: To compare the efficacy and safety of clindamycin phosphate vaginal cream with that of triple sulfonamide vaginal cream in the treatment of bacterial vaginosis. STUDY DESIGN: In this double-blind, randomized multicenter study, nonpregnant women 16 years of age or older with symptomatic bacterial vaginosis were assigned to receive either 2% clindamycin phosphate vaginal cream or triple sulfonamide vaginal cream for 7 days. Follow-up visits were conducted 5 to 10 days and 25 to 39 days after completion of treatment. RESULTS: Clinical cure or improvement at 25 to 39 days was noted in 55 (69.6%) of 79 assessable participants who received clindamycin vaginal cream and in 33 (41.8%) of 79 women who received triple sulfonamide vaginal cream (P < 0.0001). Most of the difference between the treatment groups was noted in women with a history of bacterial vaginosis. Among women without a history of bacterial vaginosis, clindamycin and triple sulfonamide creams had similar efficacy. Evaluation of Gram-stained vaginal smears correlated with clinical outcome. Most patients in both treatment groups reported an improvement in symptoms. No significant difference was observed between the treatment groups in the incidence of adverse events. CONCLUSION: Clindamycin 2% vaginal cream is more effective than triple sulfonamide vaginal cream in the treatment of bacterial vaginosis.

Single-dose gatifloxacin compared with ofloxacin for the treatment of uncomplicated gonorrhea: a randomized, double-blind, multicenter trial.

BACKGROUND: Treatment of gonorrhea is complicated by widespread resistance of Neisseria gonorrhoeae to antimicrobial agents of choice, including decreased susceptibility to ciprofloxacin. GOAL: To demonstrate the efficacy and safety of gatifloxacin, a novel 8-methoxy fluoroquinolone antibiotic, compared with ofloxacin in treating patients with uncomplicated gonococcal infection. STUDY DESIGN: In a double-blind, randomized (2:2:1), controlled trial, 340 men and 388 women with uncomplicated gonorrhea who were 16 years or older received a single oral dose of gatifloxacin (400 mg or 600 mg) or ofloxacin (400 mg). Primary analysis of efficacy was based on bacteriologic eradication from sites of infection. Secondary analyses examined clinical response and adverse event profiles. RESULTS: Bacteriologic eradication rates for gatifloxacin in evaluable men with urethral gonorrhea were 99% (400 mg) and 100% (600 mg) versus 100% for ofloxacin (n = 117, 122, and 55, respectively; P = ns). Eradication rates in evaluable women with endocervical gonorrhea were 99% for both 400 mg and 600 mg gatifloxacin versus 100% for ofloxacin (n = 101, 104, and 55, respectively; P = ns). Eradication rates were 100% for both rectal (n = 43) and pharyngeal (n = 31) infection across all treatment groups. All three drug regimens were well tolerated and exhibited similar clinical response profiles. CONCLUSION: Gatifloxacin is safe and effective as a single 400-mg or 600-mg dose for the treatment of uncomplicated gonorrhea. Similar efficacy rates were observed with the 400-mg and 600-mg doses. A single 400-mg dose can be recommended for treatment of uncomplicated gonorrhea.

HIV infection with negative serological tests: development of seropositivity in association with highly active antiretroviral therapy.

A 19-year-old woman with well-documented HIV-1 infection had persistently negative enzyme immunoassay (EIA) and Western blot serological tests. She has plasma HIV-1 RNA levels of > 480,000 copies/mL and T-helper cell counts of approximately 100/mm3. When treated with highly active antiretroviral therapy (HAART), the viral load became undetectable (< 400 copies/mL), the T-helper cell count increased to > 500/mm3 and EIA and Western blot tests became positive.

Ability of the digene hybrid capture II test to identify Chlamydia trachomatis and Neisseria gonorrhoeae in cervical specimens.

The Digene Hybrid Capture II (HCII CT/GC) test is a combination test designed to detect Chlamydia trachomatis and Neisseria gonorrhoeae in a single specimen. It is a nucleic acid hybridization test which uses signal amplification to increase sensitivity. We compared its performance to that of culture on cervical specimens from 1,370 women. Direct fluorescent-antibody assay was used to resolve discrepant results for C. trachomatis. Samples were collected with a proprietary cervical brush or with endocervical swabs. The HCII CT/GC test proved to be sensitive and specific in detecting these organisms. Compared to N. gonorrhoeae culture, it had a sensitivity of 93% (87/94) and a specificity of 98.5% (1,244/1,263). Compared to C. trachomatis culture, the sensitivity was 97.7% (129/132) and specificity was 98.2% (1,216/1,238). Testing of some specimens with discrepant results by PCR suggested that the test would actually prove to be even more specific if it were compared to a nucleic acid amplification test (NAAT). The sensitivity of C. trachomatis culture was somewhat less, at 88.6% (117/132). The endocervical brush appeared to be better than Dacron swabs for collecting specimens. The HCII CT/GC test offers an attractive format that allows simultaneous detection of C. trachomatis and N. gonorrhoeae with a single specimen. An initial positive result is followed by repeat tests with probes to identify chlamydiae or gonococci. This test is more sensitive than C. trachomatis culture and is at least as sensitive as culture for gonococci. It deserves further evaluation and comparison with NAATs and may well offer an attractive alternative for diagnosis and screening of these infections.

Evaluation of the surgical treatment of vulvar vestibulitis.

OBJECTIVE: To assess the efficacy of perineoplasty in the management of vulvar vestibulitis. STUDY DESIGN: Forty-two women who had undergone operative perineoplasty for the treatment of vulvar vestibulitis completed a questionnaire, a mean of 4.8 years postoperatively. RESULTS: Vulvodynia was constant or daily in 29 (69%) before surgery and in eight (19%) of respondents after surgery. In all, 27 (80%) of 34 women who had preoperative vulvar discomfort reported that the discomfort was much better or absent following surgery. Before surgery, 26 (70%) of 37 women who were not celibate for reasons other than vulvar vestibulitis, were celibate because of vulvar vestibulitis or always had pain during coitus and sometimes had to discontinue coitus because of pain. In contrast, only two (5.7%) of 35 women had this degree of dyspareunia following surgery. Similarly, 28 (85%) of 33 sexually active women who had dyspareunia before surgery reported that intercourse was much less painful or pain-free following surgery. CONCLUSION: Perineoplasty has a role in the management of vulvar vestibulitis for women who do not achieve satisfactory relief of vulvodynia and/or dyspareunia with nonoperative treatments.

Double-blind comparison of trovafloxacin and doxycycline in the treatment of uncomplicated Chlamydial urethritis and cervicitis. Trovafloxacin Chlamydial Urethritis/Cervicitis Study Group.

BACKGROUND: Chlamydia trachomatis is among the most common sexually transmitted bacteria worldwide. With excellent activity against C. trachomatis and Neisseria gonorrhoeae and prolonged elimination half-life allowing once-daily dosage, the fluoroquinolone trovafloxacin has potential advantages in the treatment of uncomplicated chlamydial infection. GOAL OF THIS STUDY: This study compared the efficacy of trovafloxacin with that of doxycycline for the treatment of uncomplicated chlamydial infection. STUDY DESIGN: In a double-blind, multicenter trial, trovafloxacin 200 mg was administered once daily for 5 days and doxycycline 100 mg was administered twice daily for 7 days to patients with uncomplicated chlamydial urethritis or cervicitis. Follow-up visits were conducted 10, 21, and 35 days after enrollment. RESULTS: Of the 970 patients (403 men, 567 women) observed, 511 were microbiologically evaluable and 360 were clinically evaluable. C. trachomatis eradication rates in the trovafloxacin and doxycycline groups were equivalent in women (95% and 97%, respectively), but not in men (89% and 99%). Similarly, rates of clinical success (cure plus improvement) demonstrated equivalence of trovafloxacin and doxycycline in women (96% and 94%), but not in men (94% and 100%). The most frequent treatment-related adverse events were dizziness, nausea, and headache in patients given trovafloxacin, and nausea, vomiting, and headache in patients given doxycycline. Treatment-related discontinuations were comparable between the drug groups. CONCLUSION: Trovafloxacin given once daily for 5 days was clinically and bacteriologically equivalent to doxycycline given twice daily for 7 days in women with uncomplicated chlamydial cervicitis. This equivalence was not demonstrated in men with uncomplicated chlamydial urethritis.

Bacterial vaginosis is not associated with circumcision status of the current male partner.

OBJECTIVE: Bacterial vaginosis (BV) is common in sexually active women, and in a large proportion the underlying aetiology is unknown. We evaluated partner circumcision status as a potential risk and hypothesised that women with uncircumcised partners were at increased risk for BV. METHODS: Retrospective audit of a partner study (272 heterosexual couples) conducted in Baltimore between 1990 and 1992. BV defined by clinical criteria and circumcision status of males was determined by physical examination. RESULTS: BV was diagnosed in 83 (30%) female partners; 75 (27%) males were uncircumcised. In males and females respectively, gonorrhoea was diagnosed in 20% and 16%, and chlamydia in 7% and 11%. In women with circumcised partners, 58/197 (29%) had BV compared with 25/75 (33%) with uncircumcised partners (p = 0.53). CONCLUSION: Women with uncircumcised current partners are not at increased risk for BV.

Daily oral grepafloxacin vs. twice daily oral doxycycline in the treatment of Chlamydia trachomatis endocervical infection.

OBJECTIVE: To compare the efficacy and safety of a 7-day course of treatment with oral grepafloxacin, 400 mg once daily, and oral doxycycline, 100 mg twice daily, in patients with chlamydial cervicitis. METHODS: Women aged 18 years or older attending 17 sexually transmitted disease clinics in the United States who had clinical signs of mucopurulent cervicitis or who had a recent positive culture or nonculture test for Chlamydia trachomatis or who had contact with a male partner with a positive culture for C. trachomatis were enrolled into this randomized, double-blind, active-controlled clinical study. The diagnosis of chlamydial cervicitis was based on culture for C. trachomatis. Patients were randomized to receive a 7-day course of treatment with either oral grepafloxacin, 400 mg once daily, or oral doxycycline, 100 mg twice daily. Response to therapy was assessed 3-8 days and 21-28 days after completion of treatment. The primary measure of efficacy was eradication of C. trachomatis at the 21-28 day follow-up visit. Clinical success, defined as improvement or complete resolution of the signs and symptoms of cervicitis, was a secondary measure of efficacy. RESULTS: Of the 451 female patients enrolled, 228 received grepafloxacin and 223 received doxycycline. In all, 154/451 (35%) patients were evaluable at the 21-28 day follow-up (81 who received grepafloxacin and 73 who received doxycycline). Microbiologic and clinical success rates demonstrated the equivalence of the two treatments. The C. trachomatis eradication rates were 96.3% (78/81) and 98.6% (72/73) for patients receiving grepafloxacin or doxycycline, respectively. The two study drugs were also equivalent in resolving clinical signs and symptoms, with clinical success rates of 88.9% (64/72) and 89.5% (51/57) for patients treated with grepafloxacin and doxycycline, respectively. Both drugs were well tolerated, with 47% of patients receiving grepafloxacin and 46% of patients receiving doxycycline experiencing drug-related adverse events, none of which was serious. CONCLUSIONS: Seven days of treatment with oral grepafloxacin, 400 mg once daily, was as effective as 7 days of treatment with oral doxycycline, 100 mg twice daily, in patients with cervicitis caused by C. trachomatis. Both agents were well tolerated and had comparable safety profiles. Grepafloxacin's once-daily dosing regimen may offer advantages in terms of patient compliance.

A pilot study of metronidazole vaginal gel versus oral metronidazole for the treatment of Trichomonas vaginalis vaginitis.

BACKGROUND: Trichomonas vaginalis is a common sexually transmitted pathogen. In the United States, oral metronidazole is the only officially sanctioned treatment option. OBJECTIVE: This study was undertaken to compare the efficacy and safety of 0.75% metronidazole vaginal gel with that of oral metronidazole for the treatment of trichomonal vaginitis. STUDY DESIGN: Women with trichomoniasis were enrolled in this randomized, open-label pilot study of 0.75% metronidazole vaginal gel twice daily for 7 days compared with 7 days of generic oral metronidazole, 250 mg, three times daily. Patients were seen for follow-up visits 5 to 7 days and 21 to 28 days after the last dose of medication. RESULTS: Using culture for test of cure, trichomonal infection was eliminated in all 15 women treated with oral metronidazole and 7 (44%) of 16 women treated with intravaginal metronidazole. Adverse events were similar, except that there were more taste-related adverse events in the oral metronidazole group. Significant reductions in genitourinary symptoms were seen in both the oral and intravaginal groups. CONCLUSION: This study has shown that 0.75% metronidazole vaginal gel is not effective as a single agent for the treatment of trichomoniasis. Future studies may define a role for metronidazole gel for symptomatic relief in patients intolerant of oral medication or as adjunctive treatment with oral metronidazole for the management of patients infected with metronidazole-resistant strains of T. vaginalis.

Sexual behavior and contraceptive use. Changes from 1975 to 1995 in college women.

OBJECTIVE: The objective of this study was to compare the sexual practices and contraceptive use in a sample of college women in 1995 with women surveyed in 1975, 1986 and 1989. STUDY DESIGN: We surveyed 336 college women seen at a university student health service or on campus and compared their responses to those of women surveyed at the university in 1975, 1986 and 1989. RESULTS: The proportions of women who were sexually experienced, number of life-time male sexual partners, number of male sexual partners in the past year and frequencies of specific sexual practices were similar over the four survey times. Condom use was reported as the usual method of contraception in 7% of sexually experienced women in 1975, 14% in 1986, 25% in 1989 and 46% in 1995 (P < .00001, linear trend). CONCLUSION: We found little change in sexual practices in this college population over the four survey years, with the exception of an increase in the self-reported use of condoms. Increased educational efforts should emphasize safe sexual practices (barrier methods) to prevent sexually transmitted diseases and highly efficacious methods of contraception (hormonal contraception) to avoid unintended pregnancy.

Trichomoniasis in a postmenopausal woman cured after discontinuation of estrogen replacement therapy.

BACKGROUND: Epidemiologic and experimental data suggest that estrogen has a salutary effect on Trichomonas vaginalis. CASE: A metronidazole-allergic postmenopausal woman was cured of vaginal trichomoniasis in association with discontinuation of estrogen replacement therapy. CONCLUSION: Hormonal manipulation should be studied for the management of women with trichomoniasis who are allergic to metronidazole or who are infected with metronidazole-resistant strains of Trichomonas vaginalis.

Comparison of single-dose oral grepafloxacin with cefixime for treatment of uncomplicated gonorrhea in men. The STD Study Group.

In a randomized open study, 351 male patients with uncomplicated gonorrhea were given single oral doses of grepafloxacin (400 mg) or cefixime (400 mg). In the 299 microbiologically evaluable patients, urethral infections were cured in 99% (147 of 149) of those receiving grepafloxacin and 97% (145 of 150) of those given cefixime. Eradication rates for both regimens were 100% in the 16% (47 of 299) of participants who were infected with penicillin-resistant Neisseria gonorrhoeae and 97% in the 21% (62 of 299) of participants infected with tetracycline-resistant strains. Grepafloxacin is a well-tolerated alternative to cefixime for treatment of uncomplicated gonorrhea in males.

Multicenter comparison of clotrimazole vaginal tablets, oral metronidazole, and vaginal suppositories containing sulfanilamide, aminacrine hydrochloride, and allantoin in the treatment of symptomatic trichomoniasis.

BACKGROUND AND OBJECTIVES: Trichomonas vaginalis is a common vaginal pathogen. Oral metronidazole is the drug of choice for the treatment of trichomoniasis. Oral metronidazole, however, may cause unpleasant side effects and is contraindicated during the first trimester of pregnancy. In vitro studies and preliminary clinical data have suggested that intravaginal clotrimazole may be effective against this pathogen. GOALS: To compare the efficacy of clotrimazole vaginal tablets, oral metronidazole, and vaginal suppositories containing sulfanilamide, aminacrine, and allantoin (AVC suppositories) in the treatment of women with symptomatic trichomoniasis. STUDY DESIGN: In a multicenter, open-label trial conducted in 1982 and 1983, 168 symptomatic women with microscopically evident vaginal trichomoniasis were randomized to receive any of 2 g of metronidazole as a single oral dose, two 100-mg clotrimazole vaginal tablets once a day for 7 days, or vaginal suppositories containing 1.05 g of sulfanilamide, 14 mg of aminacrine hydrochloride, and 140 mg of allantoin (AVC suppositories) twice a day for 7 days. Wet mounts and cultures were repated at 1 to 2 and 4 to 6 weeks after completion of treatment. RESULTS: The number of patients who had positive cultures after treatment were 40/45 (88.9%) in the clotrimazole group, 35/43 (81.4%) in the AVC suppository group, and 9/45 (20%) in the metronidazole group (P < 0.001). All treatments were associated with a reduction in reported symptoms. Oral metrohidazole was more effective in reducing symptoms than either of the topical preparations. Adverse events, mostly mild or moderate in severity, were reported by 7 (14.6%) of 48 patients who had received oral metronidazole and 4 (7.8%) of 51 women who used AVC suppositories. There were no adverse events reported by the 50 women who used clotrimazole vaginal tablets. CONCLUSIONS: Oral metronidazole was more effective in eradicating T. vaginalis than clotrimazole vaginal tablets or AVC vaginal suppositories. All three regimens reduced symptoms; oral metronidazole was more effective in reducing symptoms than either topical preparation.

Grepafloxacin versus cefixime as single-dose therapy for uncomplicated gonorrhea in women.

OBJECTIVE: To compare the efficacy and tolerance of single-dose grepafloxacin with cefixime for treatment of uncomplicated gonorrhea in women. METHODS: Women attending nine sexually-transmitted-disease clinics in the United States who had suspected uncomplicated gonorrhea were enrolled in an open study. Participants were randomized to receive single oral doses of grepafloxacin (400 mg) or cefixime (400 mg), and efficacy was evaluated in those who returned for follow-up assessment 5 to 10 days later. The primary measure of efficacy was microbiological response to therapy as determined by pre- and posttreatment culture results for Neisseria gonorrhoeae. RESULTS: Of 380 patients enrolled, 124 in the grepafloxacin group and 131 in the cefixime group were evaluated for microbiological response. Cervical gonococcal infections were eradicated in 99% of patients in both treatment groups, with only one persistent infection in each group. All pharyngeal (n = 15) and rectal (n = 32) gonococcal infections treated with grepafloxacin were cured, whereas 5 of 16 (31%) pharyngeal and 1 of 38 (3%) rectal infections failed to respond to cefixime. Although a third (123 of 386) of N. gonorrhoeae pretreatment isolates were resistant to penicillin or tetracycline, this had no impact on cure rates. Both drugs were well tolerated, with vaginitis being the most common treatment-related adverse event in each group. CONCLUSIONS: This study shows that single-dose grepafloxacin is at least as effective as cefixime for treating women with uncomplicated cervical gonorrhea. Grepafloxacin also appears to be highly effective against extragenital infections.