Tumor Variant Identification That Accounts for the Unique Molecular Landscape of Pediatric Malignancies.

Precision oncology trials for pediatric cancers require rapid and accurate detection of genetic alterations. Tumor variant identification should interrogate the distinctive driver genes and more frequent copy number variants and gene fusions that are characteristics of pediatric tumors. Here, we evaluate tumor variant identification using whole genome sequencing (n = 12 samples) and two amplification-based next-generation sequencing assays (n = 28 samples), including one assay designed to rapidly assess common diagnostic, prognostic, and therapeutic biomarkers found in pediatric tumors. Variant identification by the three modalities was comparable when filtered for 151 pediatric driver genes. Across the 28 samples, the pediatric cancer-focused assay detected more tumor variants per sample (two-sided, P < .05), which improved the identification of potentially druggable events and matched pathway inhibitors. Overall, our data indicate that an assay designed to evaluate pediatric cancer-specific variants, including gene fusions, may improve the detection of target-agent pairs for precision oncology.

Cumulative exposure to stimulants and immune function outcomes among HIV-positive and HIV-negative men in the Multicenter AIDS Cohort Study.

We examined associations between stimulant use (methamphetamine and cocaine) and other substances (nicotine, marijuana, alcohol and inhaled nitrites) with immune function biomarkers among HIV-seropositive (HIV +) men taking highly active antiretroviral therapy (ART) and HIV-seronegative (HIV-) men in the Multicenter AIDS Cohort Study. Among HIV + men, cumulative adherence to ART (4.07, 95% confidence interval [CI]: 3.52, 4.71, per 10 years of adherent ART use), and recent cohort enrolment (1.38; 95% CI: 1.24, 1.55) were multiplicatively associated with increase in CD4+/CD8+ ratios. Cumulative use of methamphetamine (0.93; 95% CI: 0.88, 0.98, per 10 use-years), cocaine (0.93; 95% CI: 0.89, 0.96, per 10 use-years) and cumulative medical visits (0.99; 95% CI: 0.98, 0.99, per 10 visit-years), each showed small negative associations with CD4+/CD8+ ratios. Among HIV- men, cumulative medical visits (0.996; 95% CI: 0.993, 0.999), cumulative number of male sexual partners (0.999; 95% CI: 0.998, 0.9998, per 10 partner-years) and cigarette pack-years (1.10; 95% CI: 1.02, 1.18, per 10 pack-years) were associated with CD4+/CD8+ ratios over the same period. ART adherence is associated with a positive immune function independent of stimulant use, underscoring the influence of ART on immune health for HIV+ men who engage in stimulant use.

Sexual sensation seeking, reduced concern about HIV and sexual risk behaviour among gay men in primary relationships.

Gay and bisexual men who indicated they were currently in a primary relationship with another man (N = 230) completed measures of HIV treatment attitudes, sexual risk behaviour and sexual sensation seeking. Results indicate non-primary partner sexual activity is common in many gay relationships and men in non-exclusive relationships possessed greater levels of sexual sensation seeking and treatment-related reduced concern about the dangerousness of HIV than men in exclusive relationships. Results also suggest that individuals who were members of HIV-seroconcordant relationships were more likely to engage in unprotected sexual activity with their primary sexual partners than gay men who were members of HIV-discordant couples. A series of regression analyses revealed that reduced concern about HIV mediated the relationship between sexual sensation seeking and sexual risk behaviour. The next generation of HIV prevention interventions must address the attitudinal shifts that have occurred among some gay men regarding the seriousness of HIV and should be sensitive to the dynamics of gay relationships.

Expectancies of sexual "escape" and sexual risk among drug and alcohol-involved gay and bisexual men.

PURPOSE: We tested the hypotheses that sexual risk would relate to gay/bisexual men's patterns of combining alcohol or drugs with sex, their motivation to use drugs to cognitively "escape" awareness of HIV risk, and their use of bars as social and sexual settings. METHODS: We conducted extensive interviews among African-American (n = 139) and White (n = 112) gay and bisexual men who were attending a behavioral intervention for safer sex results. Those who frequently combined drugs with sex reported higher rates of sexual risk and Hepatitis B infection than did men who infrequently combined substances with sex, or who combined only alcohol with sex. Sexual risk was pronounced among more frequent drug users who also reported strong expectancies that alcohol or drugs facilitate sex and cognitively escape from awareness of HIV risk. Frequenting bars per se was not an important factor in sexual risk. IMPLICATIONS: Men who use alcohol or drugs to enhance sexuality and escape self-awareness of HIV risk have a significantly diminished capacity to avoid sexual risk.

Recombinant human interleukin-6 enhances the immunoglobulin secretion of a rabbit-rabbit hybridoma.

The generation of stable rabbit-rabbit hybridomas is now possible by the recent development of a rabbit fusion partner. The ability to generate rabbit monoclonal antibodies (MAbs) can be advantageous because these rabbit immunoglobulins tend to exhibit higher affinity than murine MAbs. Furthermore, it has been observed that, in general, rabbits will elicit an immune response to antigens of limited immunogenicity in mice. Unfortunately, these rabbit-rabbit hybridomas secrete only 200 ng/mL to 5 microg/mL of immunoglobulin, which may limit larger scale production of rabbit antibodies. This study sought to determine if interleukin 6 (IL-6), which has been reported to have proliferative and secretory stimulating effects on some murine hybridomas, had any effect on a rabbit cell line that secretes a monoclonal IgG specific for estradiol. The results demonstrated that recombinant human IL-6 had a dose-dependent enhancing effect on the IgG secretion of the rabbit-rabbit hybridoma. The enhancing effect was consistent when the cells were continuously passed in the presence of IL-6. However, IL-6 did not affect the growth of the hybridoma. In contrast, no discernible effect was accomplished with recombinant mouse IL-6. Furthermore, no basal IL-6 activity was detected in the rabbit hybridoma extracellular medium. The IL-6 enhancement effect observed in this study may help to increase the immunoglobulin yield of rabbit hybridomas and to assist in the understanding of the mechanism(s) behind the lowered secretion level.

Risk of psychiatric disorders among individuals reporting same-sex sexual partners in the National Comorbidity Survey.

OBJECTIVES: This study examined the risk of psychiatric disorders among individuals with same-sex sexual partners. METHODS: Data are from the National Comorbidity Survey, a nationally representative household survey. Respondents were asked the number of women and men with whom they had sexual intercourse in the past 5 years. Psychiatric disorders according to Diagnostic and Statistical Manual of Mental Disorders, Revised Third Edition (DSM-III-R) criteria were assessed with a modified version of the Composite International Diagnostic Interview. RESULTS: A total of 2.1% of men and 1.5% of women reported 1 or more same-sex sexual partners in the past 5 years. These respondents had higher 12-month prevalences of anxiety, mood, and substance use disorders and of suicidal thoughts and plans than did respondents with opposite-sex partners only. Decomposition showed that the elevated same-sex 12-month prevalences were largely due to higher lifetime prevalences. Ages at onset and persistence of disorders did not differ between the same-sex and opposite-sex subsamples. CONCLUSIONS: Homosexual orientation, defined as having same-sex sexual partners, is associated with a general elevation of risk for anxiety, mood, and substance use disorders and for suicidal thoughts and plans. Further research is needed to replicate and explore the causal mechanisms underlying this association.

Impact of combination therapies on HIV risk perceptions and sexual risk among HIV-positive and HIV-negative gay and bisexual men.

The availability of improved HIV treatments may prompt reduced concern about HIV and sexual risk. Gay and bisexual men (N = 554, 17% HIV-positive) completed measures of treatment attitudes, sexual risk, and assumptions regarding the infectiousness of sexual partners. A substantial minority reported reduced HIV concern related to treatment advances. Reduced HIV concern was an independent predictor of sexual risk, particularly among HIV-positive men. In response to hypothetical scenarios describing sex with an HIV-positive partner, participants rated the risk of unprotected sex to be lower if the partner was taking combination treatments and had an undetectable viral load, relative to scenarios with a seropositive partner not taking combination treatments. Prevention efforts must address attitudinal shifts prompted by recent treatment successes, stressing the continued importance of safer sex, and that an undetectable viral load does not eliminate infection risks.

The determination of relevant goals and criteria used to select an automated patient care information system: a Delphi approach.

OBJECTIVES: To determine the relevant weighted goals and criteria for use in the selection of an automated patient care information system (PCIS) using a modified Delphi technique to achieve consensus. DESIGN: A three-phase, six-round modified Delphi process was implemented by a ten-member PCIS selection task force. The first phase consisted of an exploratory round. It was followed by the second phase, of two rounds, to determine the selection goals and finally the third phase, of three rounds, to finalize the selection criteria. RESULTS: Consensus on the goals and criteria for selecting a PCIS was measured during the Delphi process by reviewing the mean and standard deviation of the previous round's responses. After the study was completed, the results were analyzed using a limits-of-agreement indicator that showed strong agreement of each individual's responses between each of the goal determination rounds. Further analysis for variability in the group's response showed a significant movement to consensus after the first goal-determination iteration, with consensus reached on all goals by the end of the second iteration. CONCLUSION: The results indicated that the relevant weighted goals and criteria used to make the final decision for an automated PCIS were developed as a result of strong agreement among members of the PCIS selection task force. It is therefore recognized that the use of the Delphi process was beneficial in achieving consensus among clinical and nonclinical members in a relatively short time while avoiding a decision based on political biases and the "groupthink" of traditional committee meetings. The results suggest that improvements could be made in lessening the number of rounds by having information available through side conversations, by having other statistical indicators besides the mean and standard deviation available between rounds, and by having a content expert address questions between rounds.

Recombinant mouse-human chimeric antibodies as calibrators in immunoassays that measure antibodies to Toxoplasma gondii.

In the present study, we examined the feasibility of using recombinant antibodies containing murine variable regions and human constant regions as calibrators or controls in immunoassays. As a model system, we chose the Abbott IMx Toxo immunoglobulin M (IgM) and Toxo IgG assays designed to detect antibodies to Toxoplasma gondii. Two mouse monoclonal antibodies were selected based on their reactivity to the T. gondii antigens P30 and P66. Heavy- and light-chain variable-region genes were cloned from both hybridomas and transferred into immunoglobulin expression vectors containing human kappa and IgG1 or IgM constant regions. The constructs were stably transfected into Sp2/0-Ag14 cells. In the IMx Toxo IgG assay, immunoreactivity of the anti-P30 chimeric IgG1 antibody paralleled that of the positive human plasma-derived assay calibrators. Signal generated with the anti-P66 chimeric IgG1 antibody was observed to plateau below the maximal reactivity observed for the assay calibrator. Examination of the IgM chimeric antibodies in the IMx Toxo IgM assay revealed that both the anti-P30 and anti-P66 antibodies matched the assay index calibrator manufactured with human Toxo IgM-positive plasma. When evaluated with patient samples, the correlation between results obtained with the chimeric antibody calibrators and the positive human plasma calibrators was > or =0.985. These data demonstrate that chimeric mouse-human antibodies are a viable alternative to high-titer positive human plasma for the manufacture of calibrators and controls for diagnostic assays.

Sex, drugs and escape: a psychological model of HIV-risk sexual behaviours.

The lack of an effective HIV vaccine or other biomedical intervention means that behavioural change will continue to be critical to the prevention of HIV infection. Despite near universal knowledge of HIV and sexual safety, and widespread intentions to be safe, rates of unprotected sex and HIV sero-conversion remain high among gay and bisexual men. Explanatory models that link risk-taking and prevention to rational processes such as knowledge, social norms, behavioural intentions, or perceived vulnerability to infection, cannot fully account for the continued risk behaviours observed in virtually all cohorts of gay men. We feel that innovative conceptions of risk and risk prevention are needed, that emphasize non-rational, affective processes in risk-taking and decision-making. Consistent with recent models from social psychology, we propose that for many people sexual risk does not stem from a lack of community norms or personal standards, but from a desire to escape cognitive awareness of very rigorous norms and standards. Being self-aware of HIV risk arouses anxiety and precludes highly-desired activities: fatigue, fatalism, or other negative affect over HIV may lead people to 'cognitively disengage' within the sexual situation, and not to follow their norms or intentions toward safety. We propose that both substance use and the approach of high stimulation or other sexual settings facilitates this cognitive disengagement, wherein people enact 'automatic' sexual scripts and/or become more responsive to external pressures toward risk. We briefly review current psychosocial models of HIV risk behaviour, outline a cognitive escape model with particular emphasis on substance use as a behavioural risk factor, and discuss implications of an escape model for behavioural interventions among gay and bisexual men.

Sexual adventurism, high-risk behavior, and human immunodeficiency virus-1 seroconversion among the Chicago MACS-CCS cohort, 1984 to 1992. A case-control study.

BACKGROUND AND OBJECTIVES: To predict incident human immunodeficiency virus (HIV)-1 seroconversions among a cohort of gay and bisexual men based on recalled sexual behavior, drug use, partnership status, and an index of sexual adventurism/risk-seeking attitudes. STUDY DESIGN: A nested case-control design was used in a retrospective study spanning a 9-year period. RESULTS: Sexual adventurism was an important predictor of HIV-1 infection. The partial risk ratio for our 100-point adventurism scale indicated a marginal rate of increase in seroconversion risk of 4% (odds ratio = 1.04; 95% confidence interval = 1.02 to 1.06), with almost 79% of seroconverters scoring above the median on the index. As expected, partner status, drug use, and unprotected receptive anal (RA) intercourse were associated with seroconversion. However, multivariate results indicated that men using condoms consistently in RA sex were also at higher risk for infection (odds ratio = 2.68; 95% confidence interval = 1.04 to 6.95) than men who abstained from RA intercourse. CONCLUSIONS: Results are discussed with respect to their implications for HIV prevention intervention and research. Recommendations include the development of new approaches and the adaptation of existing intervention techniques that recognize and treat persons with strong risk-taking predispositions.

Substance use, HIV, and gay men.

AIDS: Recreational drug use is linked to infectious disease transmission among gay men. A review of the literature on substance use and sexual behavior divided the research into three categories: studies that relate sexual behavior to global measures of drug use; studies that look within groups of gay men at drug use behavior in the sexual context; and studies that look at sexual episodes among individual cases to determine whether the use of specific drugs lead to specific sexual activities. The evidence for a direct association between substance use and increased risk of HIV infection among susceptible gay men is incontrovertible. Simple associational models demonstrate the ways in which the pairing of sex and drug use create a learned association between these activities. The drug-sex relationship may also be mediated by personality characteristics, such as sensation-seeking. Interventions targeting gay men must combine community interventions and individual or small group intervention approaches.^ieng

Treatment of progressive rheumatoid interstitial lung disease with cyclosporine.

A 62-year-old man with longstanding rheumatoid arthritis (RA) presented with dyspnea. Active rheumatoid interstitial lung disease was documented by high resolution computed tomography, gallium scan, and bronchoalveolar lavage. He responded to high dose prednisone, but had unacceptable side effects. Chlorambucil and cyclophosphamide were not steroid sparing. After starting cyclosporine 3 mg/kg/day he was able to stop prednisone and his symptoms improved and stabilized. Pulmonary function showed sustained improvement during 2 years of followup. His RA has been well controlled. Side effects have been mild hypertension and increased serum creatinine.

A case-control study of human immunodeficiency virus type 1 seroconversion and risk-related behaviors in the Chicago MACS/CCS Cohort, 1984-1992. Multicenter AIDS Cohort Study. Coping and Change Study.

This paper focuses on 76 human immunodeficiency virus type 1 (HIV-1) seroconverters who concurrently participated in the Chicago, Illinois, component of the Multicenter AIDS Cohort Study (MACS) and the Coping and Change Study (CCS) of homosexual/bisexual men between 1984 and 1992. A nested case-control analysis was performed to assess the critical behavioral risk factors associated with incident HIV-1 infection and the consistency of these relations in early (1984-1988) versus later (1989-1992) phases of the study. Univariate results revealed strong early period associations between seroconversion and various measures of receptive anal intercourse (RAI) that became considerably weaker in the study's later period. The weaker associations reflected the overall decline in levels of RAI among the cohort during the 9 years of observation. In contrast, univariate results revealed stronger later period associations between seroconversion and measures of receptive oral intercourse and insertive anal intercourse. Subsequent multivariate testing did not support the hypothesis that receptive oral intercourse and/or insertive anal intercourse have replaced unprotected RAI as important risk behaviours in the homosexual transmission of HIV-1. In conditional logistic regression models combining intercourse measures with indices of drug and condom use, only the latter variables were consistently associated with HIV-1 seroconversion in both early and later study periods. Adjusted odds ratios (ORs) for nonuse of condoms during RAI were consistently significant throughout the study (ORs = 3.7-4.8), while adjusted odds ratios for recreational drug use variables rose dramatically during the latter half of the study (e.g., for use of cocaine, OR = 81.3 (95% confidence interval 8-824) [corrected], and for use of nitrite "poppers," OR = 9.1 (95% confidence interval 1.8-45.5)). The behavioral intervention applications of these findings, as well as their relation to data from other recent cohort studies of HIV-1 seroconversion among homosexual/bisexual men, are discussed.

Cryptogenic organizing pneumonia. A report of 25 cases and a review of the literature.

Cryptogenic organizing pneumonia (COP), also known as bronchiolitis obliterans organizing pneumonia (BOOP), is an uncommon lung disease characterized by the presence of granulation tissue within the alveolar ducts and alveoli. Because of the limited published literature on this topic and limited information on outcome we reviewed our own experience over an 8-year period and also critically evaluated the literature. We reviewed all cases of COP diagnosed from 1985 through 1992 at Vancouver General Hospital: 25 patients (14 male, 11 female) aged 20-77 years (mean, 49 yr, SD +/- 17 yr). Nine patients had myeloproliferative disorder, including 6 who had allogenic bone marrow transplants; 2 patients had connective tissue disease; and 14 patients had no underlying disease (idiopathic). Data retrieved retrospectively from clinical records included demographics, risk factors, symptoms, chest radiographs, computerized tomograms, lung function tests, therapy prescribed, and response to therapy. Symptoms included dyspnea and cough (n = 15) (60%), cough only (n = 10) (40%), and fever (n = 15) (60%). Twenty-two patients were diagnosed by open lung biopsy and 3 by transbronchial biopsy. Lung imaging showed bilateral patchy airspace consolidation or nodular opacities as the main finding in 22 patients. Pulmonary function tests showed a combined restrictive and obstructive pattern. All patients received prednisone therapy except 1 patient whose idiopathic findings resolved completely with minimal treatment. Eight patients died, including 4 of the 9 patients with myeloproliferative disorder--2 from a combination of respiratory failure due to COP and graft-versus-host disease. One of 2 patients with connective tissue disease died, and 3 of 14 patients with idiopathic COP died. COP is an uncommon condition but should be considered in patients with bilateral airspace disease, especially those who fail to respond to antibiotics for presumed pneumonia. Although pulmonary function tests and CT scan findings in conjunction with the clinical features usually suggest the diagnosis, definite confirmation usually requires either open lung biopsy or transbronchial biopsy. Histologic confirmation of the diagnosis is particularly warranted as therapy with corticosteroids is usually needed for a number of months. The prognosis is excellent with idiopathic cases but more guarded especially when COP is associated with lymphoproliferative or connective tissue disease.