Jordan Glaser

Nonhemolytic, Nonmotile Gram- Positive Rods Indicative of Bacillus anthracis

We report a 40-year-old female patient who was admitted to the hospital because of a left ovarian mass torsion. A nonhemolytic, nonmotile Bacillus, suspicious of Bacillus anthracis, was isolated from a blood culture. We discuss the evaluation that led to the final identification of the bacterium as B. megaterium.

Expansion of clonotypic T-cell populations in the peripheral blood of asymptomatic Gran Chaco Amerindians infected with HTLV-IIB

Peripheral blood mononuclear cells from asymptomatic HTLV-II-infected and uninfected Gran Chaco Amerindians were analyzed using polymerase chain reaction (PCR) for expansions of T-cell receptor (TCR) V-beta gene clonotypes. Analyses were performed using primer pairs designed to identify expanded T-cell familial clonotypes based on their unique TCR beta gene rearrangements. Of the 30 HTLV-IIB-positive samples tested, five showed evidence of V-beta clonotypic T-cell expansion. Of the five expansions, two were monoclonotypic and the remaining three were oligoclonotypic. In comparison, 30 HTLV-II-negative Amerindians showed no evidence of clonotypic T-cell expansion. Amplified DNA from one of the monoclonotypic samples was subsequently cloned and sequenced and was found to have uniform variable/ diversity/joining sequences confirming its unique monoclonal T-cell expansion. This method of detecting clonal TCR beta gene rearrangements has the advantage over traditional Southern blot techniques of being more sensitive and specific even with suboptimal specimens. The prognostic significance of clonotypic T-cell expansion in a group such as the HTLV-II-infected Gran Chaco Amerindians remains to be determined.

Increased seroreactivity to HERV-K10 peptides in patients with HTLV myelopathy.

BackgroundPreviously, we had shown that persons infected with human T-cell lymphoma leukemia virus 1 or 2 (HTLV-1 or 2) had an increased prevalence of antibodies to a peptide in the Pol protein of the retrovirus HERV-K10, homologous to a peptide in HTLV gp21 envelope protein. The prevalence rate was higher in those with myelopathy vs. non-myelopathy. We have now extended our observations to a cohort restricted to North America in whom the diagnosis of HTLV myelopathy was rigorously confirmed to also test for reactivity to another HERV-K10 peptide homologous to the HTLV p24 Gag protein.MethodsSera from 100 volunteer blood donors (VBD), 53 patients with large granular lymphocytic leukemia (LGLL), 74 subjects with HTLV-1 or 2 infection (58 non-myelopathy and 16 myelopathy) and 83 patients with multiple sclerosis (MS) were evaluated in ELISA assays using the above peptides.ResultsThe HTLV myelopathy patients had a statistically significant increased prevalence of antibodies to both HERV-K10 peptides (87.5%) vs. the VBD (0%), LGLL patients (0%), MS patients (4.8%), and the HTLV positive non-myelopathy subjects (5.2%).ConclusionThe data suggest that immuno-cross-reactivity to HERV-K10 peptides and/or transactivation of HERV-K10 expression by the HTLV Tax protein may be involved in the pathogenesis of HTLV-associated myelopathy/tropical spastic paraparesis and spastic ataxia.

Is MMTV associated with human breast cancer? Maybe, but probably not

BackgroundConflicting results regarding the association of MMTV with human breast cancer have been reported. Published sequence data have indicated unique MMTV strains in some human samples. However, concerns regarding contamination as a cause of false positive results have persisted.MethodsWe performed PCR assays for MMTV on human breast cancer cell lines and fresh frozen and formalin fixed normal and malignant human breast epithelial samples. Assays were also performed on peripheral blood mononuclear cells from volunteer blood donors and subjects at risk for human retroviral infections. In addition, assays were performed on DNA samples from wild and laboratory mice. Sequencing of MMTV positive samples from both humans and mice were performed and phylogenetically compared.ResultsUsing PCR under rigorous conditions to prevent and detect “carryover” contamination, we did detect MMTV DNA in human samples, including breast cancer. However, the results were not consistent and seemed to be an artifact. Further, experiments indicated that the probable source of false positives was murine DNA, containing endogenous MMTV, present in our building. However, comparison of published and, herein, newly described MMTV sequences with published data, indicates that there are some very unique human MMTV sequences in the literature.ConclusionWhile we could not confirm the true presence of MMTV in our human breast cancer subjects, the data indicate that further, perhaps more traditional, retroviral studies are warranted to ascertain whether MMTV might rarely be the cause of human breast cancer.

Delayed Seroconversion to HTLV-II is Associated With a Stop-Codon Mutation in the pol Gene

A known HIV-1-positive intravenous drug user was found to be human T cell lymphoma/leukemia virus-II (HTLV-II) DNA positive by polymerase chain reaction but seronegative in a screening ELISA. He was consistently DNA positive but took 2 years to fully seroconvert. Sequencing of the HTLV-II strain in his cultured T lymphocytes indicated that it is a prototypical type A strain with no major differences in the long terminal repeat DNA sequence, nor major amino acid differences in the Gag, Env, Tax, and Rex proteins. However, a mutation in its pol gene created a stop codon at amino acid 543 of the Pol protein, a region that encodes for the RNase function. This mutation may account for the subjects slow seroconversion.Abstract A known HIV-1-positive intravenous drug user was found to be human T cell lymphoma/leukemia virus-II (HTLV-II) DNA positive by polymerase chain reaction but seronegative in a screening ELISA. He was consistently DNA positive but took 2 years to fully seroconvert. Sequencing of the HTLV-II strain in his cultured T lymphocytes indicated that it is a prototypical type A strain with no major differences in the long terminal repeat DNA sequence, nor major amino acid differences in the Gag, Env, Tax, and Rex proteins. However, a mutation in its pol gene created a stop codon at amino acid 543 of the Pol protein, a region that encodes for the RNase function. This mutation may account for the subjects slow seroconversion.

Flaws in Patient Safety Measures

lence of depression among unselected homeless adults is reported to range from 0% to 49%.1 A higher prevalence among homeless individuals experiencing mental illness is therefore not surprising. Participants with psychosis or bipolar disorder were more likely to be classified as high needs according to our study protocol and assigned to another group in the trial, whereas our article reported outcomes among participants with moderate needs. This assignment process caused individuals with a diagnosis of depression to represent a larger proportion of participants in the moderate-needs group (59%) than in the high-needs group (43%). Drs Dettling and Anghelescu reference a meta-analysis suggesting that staff without clinical training reported higher rates of depression among homeless adults compared with their clinically trained counterparts.1 Although not all our research staff were clinically trained, they were supervised by clinicians and received extensive training in the administration of the Mini-International Neuropsychiatric Interview (MINI), specifically designed as a quick diagnostic tool to be used by professional interviewers in clinical and research settings, including multicenter trials.2 The interviewer-rated MINI has been shown to have good specificity for diagnosis of major depressive disorder compared with the Structured Clinical Interview for DSM-III-R (0.88), the Composite International Diagnostic Interview (0.79), and expert diagnoses (0.84).2 We agree that alcohol and substance use problems are pervasive among homeless adults, and other studies have investigated the effect of substance use on study outcomes, as well as the effect of the intervention on substance use. For example, Palepu et al3 found that concurrent substance dependence did not reduce the effectiveness of Housing First interventions in improving housing stability among participants at the At Home/Chez Soi study site in Vancouver, British Columbia, Canada. At the Toronto, Ontario, Canada site, Housing First interventions resulted in greater reductions in both the number of days experiencing alcohol problems and the amount of money spent on alcohol at 24 months compared with participants in the usual care group.4 Forthcoming cross-site analyses will examine predictors of outcomes in both the highand moderate-needs groups, as well as factors associated with poor responses to the intervention. With regard to adherence to drug treatment, it is important to note that Housing First is a participant-driven intervention and does not require adherence to psychiatric medications or abstinence from drug or alcohol use.5 Substance use3 does not preclude successful housing outcomes. Furthermore, comparisons of Housing First and traditional treatment first programs have suggested better housing outcomes for the former.6 We agree that measuring quality-of-life outcomes among people with mental illness can be challenging, which is why we used 2 instruments specifically developed for individuals with mental disorders, the Lehman Quality of Life Interview 20 and the Multnomah Community Ability Scale, as exploratory outcomes in addition to the EuroQoL 5 Dimensions health questionnaire.

Using Clinical Pathways to Assess Interventions to Prevent COPD Readmissions

Two recent articles and the accompanying editorial in CHEST (May 2015) 1 3 correctly point out that 30-day readmissions aft er COPD are usually not for COPD exacerbations and that the eff ect of currently identifi ed interventions may not vary from control groups. Risk predictors for 30-day readmissions aft er COPD may depend on the principal discharge diagnosis of the readmission, since the majority of readmissions are not for COPD exacerbations.

[24] Estimation of genetic heterogeneity in primate T-cell lymphoma/leukemia viruses by PCR

Publisher Summary The primate T-cell lymphoma/leukemia viruses (PTLV) are a subgroup of retroviruses, including human T-cell lymphoma/leukemia viruses types I and II (HTLV-I and HTLV-II) and simian T-cell leukemia virus (STLV-I). The PTLV are a group of plus-sense and single-stranded RNA-containing viruses. The PTLV and bovine leukemia virus, the etiologic agent of enzootic bovine leukemia, form an unnamed genus of retroviruses. One of the characteristics of these four type C viruses is the presence of a conserved epitope in their core proteins. This epitope is absent from all other retroviruses, including human immunodeficiency virus (HIV-1) and HIV-2. Multiple isolates of each of the members of the PTLV have been characterized. The genomic diversity found among HTLV-I isolates from asymptomatic carriers and patients with adult T-cell leukemia/lymphoma and HTLV-I-associated myelopathy from throughout the world exhibits no correlation with the disease, but rather depends on their geographic origin. Polymerase chain reaction using the primer pair, SK110/SK111, has proven to be a sensitive and specific means to detect PTLV infection.