Patterns of a Rectal Microbicide Placebo Gel Use in a Preparatory Stage for a Phase I Trial Among Young Men Who Have Sex with Men.

We examined young gay, bisexual, and other men who have sex with men's (YGBMSM) usage patterns of a pre-coital, applicator-administered rectal placebo gel. An ethnically diverse sample of 94 YGBMSM (aged 18-30 years) were asked to insert hydroxyethylcellulose placebo gel rectally before receptive anal intercourse (RAI) and report their gel use through an interactive voice response system (IVRS) across 12 weeks. We used trajectory analyses to characterize participants' use of the rectal gel over the 12 weeks, and examine whether these trajectories varied based on participants' sociodemographic characteristics, sexual behaviors, application and insertion behaviors, and experiences using the placebo gel. A cubic model was the best fit for these longitudinal data, with two distinct trajectories of gel use observed. The first trajectory ('High with Varying Gel Use per Week') represented YGBMSM (N = 38; 40.3%) who reported using the rectal gel on several occasions per week. The second trajectory ('Low and Consistent Gel Use per Week') represented participants (N = 56; 59.7%) who reported a consistent average use of one gel per week. Participants in the High with Varying Gel Use Trajectory reported trying out a greater number of positions when inserting the gel across the 12-weeks than peers in the Low and Consistent Gel Use Trajectory. YGBMSM reporting more RAI occasions during the trial were more likely be present in the High with Varying Gel Use Trajectory than peers in the Low and Consistent Gel Use Trajectory. Future research examining how to facilitate gel application and adherence among YGBMSM is merited.

Anal human papillomavirus and HIV: A cross-sectional study among men who have sex with men in Moscow, Russia, 2012-2013.

Anal human papillomavirus (HPV) is prevalent among men who have sex with men (MSM), but has not been studied in the Russian Federation. A cross-sectional survey and HPV genotyping were conducted among HIV seropositive (n=58) and seronegative MSM (n=65)in Moscow. Multivariable logistic regression was performed to identify correlates of infection with oncogenic HPV genotypes 16 and/or 18 (HPV 16/18). Forty per cent (49/124) of all MSM were infected with at least one anal HPV genotype, 31.5% (39/124) had HPV16/18,and 11.5% (14/121) had high-grade squamous intraepithelial lesions (HSIL). HPV 16/18 was more prevalent in HIV seropositive than seronegative men (24/58,41.4% vs 15/65, 23.1%; p=0.03). HIV infection was independently associated with HPV 16/18 (adjusted odds ratio (AOR): 5.08; 95% confidence intervals (CI):1.49-17.34, p=0.01), as was having 2-4 steady male sex partners in the last year (vs ≤ 1 partner; AOR: 6.99;95%CI: 1.94­25.24, p<0.01). History of prison/detention,migration to/within Russia and use of incompatible lubricants were marginally associated with HPV16/18 (p<0.10). Comprehensive prevention options are needed to address HIV and HPV infection among MSM in Russia and may benefit from inclusion of young men in piloted HPV vaccination programmes.

Generation of neutralizing aptamers against herpes simplex virus type 2: potential components of multivalent microbicides.

The prophylactic use of topical antiviral agents has recently been validated by the reduction in human immunodeficiency virus (HIV) type 1 infection incidence seen using tonofovir-containing microbicides. In order to develop a wide-spectrum microbicide to prevent infection with a wide range of sexually transmitted viruses, we have previously reported the development of HIV-neutralizing aptamers and here report the isolation and characterization of aptamers that neutralize herpes simplex virus type 2 (HSV-2). These aptamers bind the envelope glycoprotein (gD), are potent (IC(50) of 20-50 nM) and are able to block infection pathways dependent on both major entry receptors, Nectin1 and HVEM. Structural analysis and mutagenesis of these aptamers reveal a core specificity element that could provide the basis for pharmaceutical development. As HSV-2 is a major risk factor for the acquisition of HIV-1, a microbicide capable of preventing HSV-2 infection would not only reduce the morbidity associated with HSV-2, but also that derived from HIV-1.

To Use a Rectal Microbicide, First Insert the Applicator: Gel and Applicator Satisfaction Among Young Men Who Have Sex With Men.

We examined how experiences with a rectal placebo gel and applicator used with receptive anal intercourse (RAI) related to young men who have sex with men's (YMSM) likelihood of using a rectal microbicide gel and applicator in the future. An ethnically diverse sample of 95 YMSM (aged 18 to 30 years) were asked to insert hydroxyethylcellulose (HEC) placebo gel rectally before RAI during 12 weeks and report the product's acceptability (i.e., satisfaction with applicator and gel, respectively; perceived gel side effects; and sexual satisfaction when gel was used) and likelihood of future microbicide use. Main and interaction effects predicting future use intentions were tested using linear regression. We found a positive association between future use intentions and applicator satisfaction (b = .33, p < .001). In a subsequent interaction effects model, we found that greater gel satisfaction was associated with increased future use intentions; however, the strength of this relationship was magnified when YMSM reported greatest satisfaction with the rectal applicator. Applicator satisfaction may be a salient factor in YMSM's decision-making to use a rectal microbicide in the future. Although the importance of developing a satisfactory rectal microbicide gel for YMSM is undeniable for its future use, our results also emphasize the importance of developing strategies that increase YMSM's comfort and skill when using a rectal applicator. Future research examining how to optimize the design, properties, and characteristics of a rectal applicator as a strategy to promote greater satisfaction and use among YMSM is merited.

Cytokine gene expression in HIV-infected intestinal mucosa.

OBJECTIVE: Cytokine dysregulation has been implicated in AIDS pathogenesis and the gastrointestinal tract, containing approximately 40% of the body's lymphoid tissue, is likely to act both as a reservoir of viral infection and a site for immune dysregulation. In this study evidence of cytokine dysregulation in intestinal mucosa has been sought using the reverse transcriptase polymerase chain reaction (RT-PCR) to amplify cytokine mRNA. METHODS: RT-PCR was performed on intestinal biopsies obtained from 50 HIV-infected patients and 31 controls. Tissue was obtained at diagnostic endoscopy and total RNA extracted using an RNAzol technique. Following RT, cDNA was amplified using primers specific for beta-actin, interleukin (IL)-1 beta, tumour necrosis factor (TNF)-alpha, interferon (IFN)-gamma, IL-2, IL-4, IL-10 and IL-13. RESULTS: There was a significant increase in the expression of the proinflammatory cytokines IL-1 beta and IFN-gamma in the HIV-infected compared with the control small intestinal samples (P < 0.01). IL-10 was significantly reduced in the respective groups' large intestine (P < 0.02). The expression of IL-2 was also reduced in both the small and large intestinal HIV samples although this was not significant. IL-13 mRNA was only detected in one control patient. CONCLUSIONS: Dysregulation of cytokine gene expression occurs in the intestinal mucosa of patients with HIV infection and is characterized by increased expression of proinflammatory cytokine mRNA. Further studies are needed to localize the cellular origin of such dysregulation and to quantify the degree of abnormality.

The natural history of cryptosporidial diarrhoea in HIV-infected patients.

OBJECTIVE: To determine the natural history of cryptosporidial infection in HIV-infected individuals. DESIGN: Retrospective study. SETTING: University teaching hospital HIV inpatient and outpatient unit. PATIENTS: Thirty-eight HIV-infected patients presenting with cryptosporidial diarrhoea between April 1986 and July 1991 were identified retrospectively from laboratory records. RESULTS: Eleven of the 38 patients had a clinical remission of their diarrhoea. Median lymphocyte count of the remission group was significantly higher than that of the non-remission group (1100 and 550 x 10(6)/l, respectively; P = 0.003). Median survival times were 66 and 11.5 weeks for the remission and non-remission groups, respectively (P = 0.001). Liver function tests performed at the initial diagnosis of cryptosporidial diarrhoea were available for 28 patients. Aspartate transaminase was raised in 16 and alkaline phosphatase in 10 of these 28 patients. Ten patients showed evidence of AIDS-associated sclerosing cholangitis, one patient had an episode of acute pancreatitis and another presented with acute cholecystitis. CONCLUSIONS: This study suggests that HIV-associated cryptosporidial diarrhoea does not have a uniformly poor prognosis. Eleven out of 38 patients had a spontaneous clinical remission, which appears to be predicted by the absolute lymphocyte count. Abnormal liver function tests and hepatobiliary disease were common.

Enhanced levels of functional HIV-1 co-receptors on human mucosal T cells demonstrated using intestinal biopsy tissue.

OBJECTIVE: To examine compartmental differences in co-receptor expression on CD4 lymphocytes between blood and gut using endoscopic biopsies. DESIGN: Mucosal and peripheral CD4 T cells from healthy controls were compared for co-receptor expression and vulnerability to infection by HIV-1. METHODS: Expression of CCR5 and CXCR4 was quantified by flow cytometry on isolated mucosal CD4 lymphocytes obtained from endoscopic biopsies and blood from healthy controls. Vulnerability to in vitro infection by both R5 and X4 strains was assessed by measuring p24. RESULTS: Biopsies yielded sufficient lymphocytes for flow cytometric characterization and infectivity studies. The percentage of mucosal CD4 T lymphocytes that expressed CCR5 and the per cell expression of CCR5 were both significantly increased compared with that in peripheral blood CD4 T lymphocytes. CXCR4 was expressed on the majority of CD4 lymphocytes in both compartments. In vitro infection of mucosal mononuclear cells supported greater viral replication of both R5 and X4 strains than peripheral blood mononuclear cells. CONCLUSIONS: Enhanced expression of CXCR4 and CCR5 on CD4 lymphocytes in normal intestinal mucosa predicts increased vulnerability to infection by both R5 and X4 HIV-1. Endoscopic biopsies provide a useful mucosal tissue sampling technique to identify compartmental immunologic differences that may be exploited by HIV-1 in establishing initial mucosal infection.

Sensitive and reproducible quantitation of mucosal HIV-1 RNA and DNA viral burden in patients with detectable and undetectable plasma viral HIV-1 RNA using endoscopic biopsies.

Mucosal tissue is the main portal of entry for HIV-1 infection and, in macaques, has been demonstrated to be a significant compartment for viral replication and CD4+ T lymphocyte depletion. Quantitating tissue viral burden in addition to plasma viral load provides insights into HIV-1 pathogenesis and an additional means to gauge antiretroviral response. The aim of this study was to develop reliable, reproducible, and sensitive assays to quantitate tissue viral burden of HIV-1 RNA and DNA using 1-3 endoscopically acquired, rectosigmoid biopsies. Total DNA and RNA were simultaneously extracted following homogenization from the same tissue samples. Quantitative polymerase chain reaction (PCR) assay in the HIV-1 LTR region was used to detect viral DNA and RT-PCR for viral RNA. It was determined that HIV-1 RNA and DNA can be reproducibly quantified from a single rectosigmoid biopsy with minimal intra-assay or intra-patient variability. These results reflect high recovery of extracted nucleic acids with calculated results accurately reflecting in vivo levels. The techniques outlined differ from currently available approaches by incorporating control standards to identify loss or degradation of RNA and DNA from acquisition through the in vitro assay and permit extraction with high yields of RNA and DNA from the same tissue sample.

Gastrointestinal mucosal biopsy in HIV disease and AIDS.

The role of the gastroenterologist as consultant for patients with HIV infection is reviewed, with a particular focus on when endoscopy with biopsy may be helpful in the diagnostic evaluation. Suggestions on where to biopsy, how to collect samples, and what pathologies might be anticipated are included. In the clinical setting of new antiviral therapies, there has been a dramatic change in the etiologic factors for common presentations such as diarrhea. A review of suspect infections and malignancies is included.

Controlled potential coulometry: the application of a secondary reaction to the determination of plutonium and uranium at a solid electrode.

A method is described for the simultaneous determination of plutonium and uranium in mixed oxides by controlled potential coulometry at a gold working electrode in two stages: first a coulometric oxidation, at 0.73 V vs. a silver/silver chloride electrode, of Pu(III) and U(IV) to Pu(IV) and U(VI) by a combination of a direct electrode reaction and a secondary chemical reaction proceeding concurrently, and secondly, a coulometric reduction at 0.33 V of Pu(IV) to Pu(III), leaving uranium as U(VI). The determination is carried out in a mixture of sulphuric and nitric acids, and Ti(III) is used to reduce plutonium and uranium to Pu(III) and U(IV) before electrolysis. The precision (3sigma) of Pu:U ratio results obtained from mixtures containing about 30% and 2% plutonium was 0.5% and 1-5% respectively. The effect of experimental variables on the time taken to complete the coulometric determination is discussed.

Determination of plutonium and uranium in mixed oxide fuels by sequential redox titration.

The use of a sequential determination of uranium and plutonium in a single sample solution results in a saving in analysis time and apparatus requirements. The method starts with U(IV) and Pu(in) in a mixture of sulphuric and nitric adds. Titration with dichromate, using amperometry at a pair of polarizable electrodes, produces two well-defined end-points corresponding to the sequential oxidation of U(IV) to U(VI) and Pu(III) to Pu(IV). The quantitative oxidation of U(IV) to U(VI) is achieved via the action of Pu(IV) as intermediate, and is dependent upon establishing conditions which favour rapid reaction between U(IV) and Pu(IV). The method is precise and accurate. With Pu-U mixtures containing between 15 and 30% plutonium the precision (3sigma) of the Pu: U ratio results is +/-0.6% on samples containing 100-120 mg of plutonium plus uranium. Iron and vanadium interfere quantitatively with plutonium, copper interferes non-quantitatively with uranium, and gross amounts of molybdenum mask the uranium end-point.

Palliative laser therapy for inoperable rectal cancer--does it work? A prospective study of quality of life.

Endoscopic neodymium yttrium aluminium garnet (Nd YAG) laser therapy, is a new and simple method for the palliative treatment of inoperable colorectal cancer. To date the authors have treated 70 patients and the value of this method was assessed prospectively in 14 patients. Quality of life was measured before, during, and after treatment. The quality of life (QL) index, a physician's assessment and a linear analogue self-assessment (LASA) were used. A close correlation was found between the two assessment methods (r = 0.79). Overall there was a significant improvement from the mean pretreatment score and the best score achieved posttreatment (QL: P = 0.002; LASA: P = 0.002). Patients with diarrhea, rectal bleeding, mucus discharge, or pain secondary to tumor bulk, will benefit most from this treatment. Malignant cachexia, pain secondary to sacral plexus involvement, tumor encroachment on the anal canal and/or sphincter dysfunction resulting from tumor invasion should be viewed as relative contraindications to laser therapy. The authors conclude that in selected patients endoscopic laser therapy can provide effective palliation in patients with malignant tumors of the rectum and descending colon.

A longitudinal psycho-physiological study of active and inactive men.

After an interval of ten years, physically active and inactive male university teachers were re-examined across four measures; predicted maximum oxygen uptake, percentage body fat, personality and attitudes towards physical activity. Both groups showed a decrease in predicted oxygen uptake and an increase in body fat although the active subjects continued to possess higher predicted maximum oxygen uptake values and have a lower percentage body fat than the inactive subjects. In general the psychological assessments revealed no major changes but minor shifts in attitudes towards physical activity were noted.

Symptomatic management of HIV associated gastrointestinal disease.

This chapter describes the differential diagnosis and management of gastrointestinal symptoms associated with HIV infection. There is no clear point when management moves from intervention to palliation, and as with other manifestations of HIV disease, clinical decisions have to be guided by the wishes of the patient. In general, early diagnosis and treatment of HIV associated opportunistic infection are likely to keep patients symptom free, but when specific therapy is unavailable, unsuccessful or unwanted, then there is a clear indication to strive for symptom control using conventional palliative care.

Advances in mucosal immunology.

HIV infection is likely to remain a significant medical and scientific problem well into the twenty-first century. During the first 15 years of the epidemic, much has been learned about the biology of HIV infection, but the majority of biomedical research has focused on the peripheral circulation. It is likely that the behavior of the virus within the unique immunologic environment of the intestinal mucosa differs from that which is observed in the periphery. Many clinical and epidemiologic features of HIV infection offer compelling reasons to encourage further examination of the mucosal immune system's role in AIDS pathogenesis. This article has touched on most of the significant observations concerning the mucosal immune system and HIV infection, and it is clear that much remains to be done. As mentioned earlier, the mucosal abnormalities observed in HIV infection are likely to have many causes. Careful evaluation of patients with early disease and fewer confounding variables may provide fresh insight into AIDS pathogenesis. Similarly, prospective evaluation of selected patient populations may be more informative in characterizing the progressive alterations in mucosal immune function than random cross-sectional studies of poorly defined groups. It is equally important for immunologic assessment to be correlated with nutritional and symptomatic evaluation. Finally, the success or failure of future antiretroviral therapies will be critically related to the impact of such agents on lymphoid reservoirs of HIV infection such as the gastrointestinal tract, which are at present refractory to treatment.

Intestinal microsporidiosis.

Intestinal microsporidiosis is caused by the protozoan parasites Enterocytozoon bieneusi and Encephalitozoon intestinalis. The disease has been described within the past decade and is found predominantly in acquired immunodeficiency syndrome (AIDS) patients in association with diarrhea. There have been rare reports of infections in immunocompetent patients. Both species of microsporidia invade and multiply within the enterocytes of the small intestine; Encephalitozoon intestinalis also causes a disseminated infection. Electron microscopy has been the mainstay of diagnosis, but improved noninvasive methods of detecting spores in feces are in development. The biology and pathogenicity of the parasites and the pathology and treatment of the disease are poorly understood.

High definition computed tomography in rheumatoid arthritis associated pulmonary disease.

Eighteen patients with rheumatoid arthritis (American Rheumatism Association definition) were selected consecutively from a rheumatology clinic. All patients were examined with plain chest radiographs, thin slice computed tomography, and pulmonary function tests. Four patients with normal chest radiographs, and normal pulmonary function tests were found to have normal CT scans. In ten patients, abnormalities consistent with rheumatoid associated lung disease were demonstrated, including changes of interstitial fibrosis in seven cases. This fibrosis had a predominantly peripheral pattern on CT scan. Computed tomography was found to be more sensitive than plain radiographs in detecting abnormalities; however, all the patients in this series with CT lung changes had abnormalities on pulmonary function testing.

Cytokine gene transcription of human colonic intraepithelial lymphocytes costimulated with epithelial cells bearing HLA-DR and its inhibition by 5-aminosalicylic acid.

The objectives of this study were to activate human colonic intraepithelial lymphocytes at the transcriptional level with HLA-DR+ human colonic epithelial cell line (HT29) in synergy with CD3 monoclonal antibody and to investigate the molecular mechanism for the therapeutic effects of 5-aminosalicylic acid. Lymphocytes were isolated by a mechanical method from resected colon of 22 cases and then cocultured on 10 ng/ml CD3mAb immobilized plates with HT29 which had been induced to express MHC class II molecules by interferon gamma. Flow cytometry analysis suggested that the lymphocyte population had a CD4/CD8 ratio similar to that observed in intact tissue sections and that there was no HT29 contamination of the lymphocytes isolated again from cocultured cells. The activation of intraepithelial lymphocytes showed the gene transcription of interferon gamma and tumor necrosis factor alpha, as measured by means of the reverse-transcriptase polymerase chain reaction, and this activation was antagonized by 5-aminosalicylic acid. Thus, epithelial cells bearing HLA-DR are capable of enhancing CD3-induced activation of human colonic intraepithelial lymphocytes and subject to inhibition by 5-aminosalicylic acid, the active moiety of salicylates used in inflammatory bowel disease.