Evaluation of RIDA®GENE norovirus GI/GII real time RT-PCR using stool specimens collected from children and adults with acute gastroenteritis.

BACKGROUND: Norovirus is the leading cause of epidemic and sporadic acute gastroenteritis (AGE) in the United States. Widespread prevalence necessitates implementation of accurate norovirus detection assays in clinical diagnostic laboratories. OBJECTIVE: To evaluate RIDA®GENE norovirus GI/GII real-time RT-PCR assay (RGN RT-PCR) using stool samples from patients with sporadic AGE. STUDY DESIGN: Patients between 14 days to 101 years of age with symptoms of AGE were enrolled prospectively at four sites across the United States during 2014-2015. Stool specimens were screened for the presence of norovirus RNA by the RGN RT-PCR assay. Results were compared with a reference method that included conventional RT-PCR and sequencing of a partial region of the 5'end of the norovirus ORF2 gene. RESULTS: A total of 259 (36.0%) of 719 specimens tested positive for norovirus by the reference method. The RGN RT-PCR assay detected norovirus in 244 (94%) of these 259 norovirus positive specimens. The sensitivity and specificity (95% confidence interval) of the RGN RT-PCR assay for detecting norovirus genogroup (G) I was 82.8% (63.5-93.5) and 99.1% (98.0-99.6) and for GII was 94.8% (90.8-97.2) and 98.6% (96.9-99.4), respectively. Seven specimens tested positive by the RGN-RT PCR that were negative by the reference method. The fifteen false negative samples were typed as GII.4 Sydney, GII.13, GI.3, GI.5, GI.2, GII.1, and GII.3 in the reference method. CONCLUSIONS: The RGN RT-PCR assay had a high sensitivity and specificity for the detection of norovirus in stool specimens from patients with sporadic AGE.

The implication of neuroactive steroids in Tourette's syndrome pathogenesis: A role for 5α-reductase?

Tourette's syndrome (TS) is a neurodevelopmental disorder characterised by recurring motor and phonic tics. The pathogenesis of TS is considered to reflect dysregulations in the signalling of dopamine (DA) and other neurotransmitters, which lead to excitation/inhibition imbalances in cortico-striato-thalamocortical circuits. The causes of these deficits may reflect complex gene × environment × sex (G × E × S) interactions; indeed, the disorder is markedly predominant in males, with a male-to-female prevalence ratio of approximately 4 : 1. Converging lines of evidence point to neuroactive steroids as being likely molecular candidates to account for G × E × S interactions in TS. Building on these premises, our group has begun examining the possibility that alterations in the steroid biosynthetic process may be directly implicated in TS pathophysiology; in particular, our research has focused on 5α-reductase (5αR), the enzyme catalysing the key rate-limiting step in the synthesis of pregnane and androstane neurosteroids. In clinical and preclinical studies, we found that 5αR inhibitors exerted marked anti-DAergic and tic-suppressing properties, suggesting a central role for this enzyme in TS pathogenesis. Based on these data, we hypothesise that enhancements in 5αR activity in early developmental stages may lead to an inappropriate activation of the 'backdoor' pathway for androgen synthesis from adrenarche until the end of puberty. We predict that the ensuing imbalances in steroid homeostasis may impair the signalling of DA and other neurotransmitters, ultimately resulting in the facilitation of tics and other behavioural abnormalities in TS.

Translational physiology and SND recordings in humans and rats: a glimpse of the recent past with an eye on the future.

The sympathetic nervous system (SNS) plays an important role in cardiovascular function, and based on the critical mechanistic relationship between altered sympathetic neural mechanisms and cardiovascular disease, it is important that the autonomic research community identifies deficiencies in the translational exchange of information and strives for a more thorough understanding of the translational significance of findings from studies involving sympathetic nerve discharge (SND) regulation in human and animal subjects. The present review assesses the state of the literature regarding studies that have used direct recordings of SND during the past three decades in humans and rats, focusing on; 1) identifying the number of studies reporting SND recordings in humans and rats, 2) briefly describing the translational exchange of SND regulation information from these studies, 3) contrasting the number of studies completed in anesthetized and conscious rats, and 4) assessing the prevalence of long-term SND recording studies in conscious rats. The majority of SND recordings in rats have been completed using anesthetized preparations, although a substantial number of studies have been completed in conscious rats. However, few studies have completed long-term (>5 days) SND recordings in freely-behaving rats, and even fewer studies have used experimental preparations that combine long-term nerve recordings with the capacity for completing central neural microinjections, or have been completed in animal models of cardiovascular disease. The wide-spread implementation of long-term SND recordings in rodent models of cardiovascular disease would be expected to enhance the translational exchange of clinically-relevant information between animals and humans.

Bacillus anthracis lethal toxin induces complex changes in sympathetic nerve discharge regulation.

Bacillus anthracis lethal toxin (LeTx) alters blood pressure and visceral sympathetic nerve discharge (SND) regulation (Garcia et al., 2012). The present results indicate that LeTx infusions produce similar response profiles in peripheral (lumbar) and visceral (renal) SND; an initial widespread activation of sympathetic nerve outflow, followed by a generalized reduction in lumbar and renal SND from peak levels, although the sympathoinhibition tended to be attenuated in lumbar SND. Combined hypoxia+hypercapnia during the hypotensive phase of LeTx infusions increased lumbar and renal SND, indicating that sympathetic neural circuits can be activated during the circulatory shock phase of B. anthracis septicemia.

Bacillus anthracis lethal toxin alters regulation of visceral sympathetic nerve discharge.

Bacillus anthracis infection is a pathophysiological condition that is complicated by progressive decreases in mean arterial pressure (MAP). Lethal toxin (LeTx) is central to the pathogenesis of B. anthracis infection, and the sympathetic nervous system plays a critical role in physiological regulation of acute stressors. However, the effect of LeTx on sympathetic nerve discharge (SND), a critical link between central sympathetic neural circuits and MAP regulation, remains unknown. We determined visceral (renal, splenic, and adrenal) SND responses to continuous infusion of LeTx [lethal factor (100 µg/kg) + protective antigen (200 µg/kg) infused at 0.5 ml/h for ≤6 h] and vehicle (infused at 0.5 ml/h) in anesthetized, baroreceptor-intact and baroreceptor (sinoaortic)-denervated (SAD) Sprague-Dawley rats. LeTx infusions produced an initial state of cardiovascular and sympathetic nervous system activation in intact and SAD rats. Subsequent to peak LeTx-induced increases in arterial blood pressure, intact rats demonstrated a marked hypotension that was accompanied by significant reductions in SND (renal and splenic) and heart rate (HR) from peak levels. After peak LeTx-induced pressor and sympathoexcitatory responses in SAD rats, MAP, SND (renal, splenic, and adrenal), and HR were progressively and significantly reduced, supporting the hypothesis that LeTx alters the central regulation of sympathetic nerve outflow. These findings demonstrate that the regulation of visceral SND is altered in a complex manner during continuous anthrax LeTx infusions and suggest that sympathetic nervous system dysregulation may contribute to the marked hypotension accompanying B. anthracis infection.

Bovine tuberculosis in Michigan wildlife.

White-tailed deer in Michigan are now recognized as a reservoir host of bovine tuberculosis (TB). It has been determined that the most likely cause of bovine TB infection in the deer is from congregating in artificially high numbers at feed sites. The presence of a wildlife reservoir of TB in Michigan poses a serious threat to the control and eradication programs that are now in their final stages in the United States.

Persistent tuberculosis or specimen contamination?

Cross-contamination during sequential processing of sputum specimens from different patients causes false-positive growth of Mycobacterium tuberculosis in culture. We describe an unusual case of cross-contamination in a 36-year-old man with acquired immunodeficiency syndrome and possible persistent tuberculosis. Culture with 1 of 3 sputum specimens was positive for rifampin-susceptible M tuberculosis. Review of processing revealed that his single culture-positive sputum specimen had followed a sputum specimen from another patient with active pulmonary tuberculosis that was positive in culture for M tuberculosis resistant to rifampin. Molecular strain typing by restriction fragment length polymorphism demonstrated the 2 isolates to be an identical strain of M tuberculosis. Agar proportion susceptibility testing of the rifampin-resistant isolate revealed low numbers of resistant organisms in a range of 1.5% to 3.3%. It was concluded that rifampin-susceptible organisms that constituted approximately 98% of the resistant isolate contaminated sputum from the patient with possible persistent tuberculosis. His culture result was, therefore, considered false positive, not an indication of tuberculosis.

Soteria and other alternatives to acute psychiatric hospitalization: a personal and professional review.

The author reviews the clinical and special social environmental data from the Soteria Project and its direct successors. Two random assignment studies of the Soteria model and its modification for long-term system clients reveal that roughly 85% to 90% of acute and long-term clients deemed in need of acute hospitalization can be returned to the community without use of conventional hospital treatment. Soteria, designed as a drug-free treatment environment, was as successful as anti-psychotic drug treatment in reducing psychotic symptoms in 6 weeks. In its modified form, in facilities called Crossing Place and McAuliffe House where so-called long-term "frequent flyers" were treated, alternative-treated subjects were found to be as clinically improved as hospital-treated patients, at considerably lower cost. Taken as a body of scientific evidence, it is clear that alternatives to acute psychiatric hospitalization are as, or more, effective than traditional hospital care in short-term reduction of psychopathology and longer-term social adjustment. Data from the original drug-free, home-like, nonprofessionally staffed Soteria Project and its Bern, Switzerland, replication indicate that persons without extensive hospitalizations (<30 days) are especially responsive to the positive therapeutic effects of the well-defined, replicable Soteria-type special social environments. Reviews of other studies of diversion of persons deemed in need of hospitalization to "alternative" programs have consistently shown equivalent or better program clinical results, at lower cost, from alternatives. Despite these clinical and cost data, alternatives to psychiatric hospitalization have not been widely implemented, indicative of a remarkable gap between available evidence and clinical practice.

Use of DNA fingerprinting to assess tuberculosis infection control.

BACKGROUND: DNA fingerprinting establishes the genetic relatedness of Mycobacterium tuberculosis isolates and has become a powerful tool in tuberculosis epidemiology. OBJECTIVE: To use DNA fingerprinting to assess the efficacy of current tuberculosis infection-control practices. DESIGN: Retrospective molecular and descriptive epidemiologic study. SETTING: A 700-bed urban public hospital that follows the Centers for Disease Control and Prevention (CDC) guidelines for tuberculosis infection control. PATIENTS: 183 patients who had positive cultures for M. tuberculosis from 1 April 1995 to 31 March 1996. RESULTS: 173 of 183 M. tuberculosis isolates from the study period underwent DNA fingerprinting. Fingerprinting revealed that five isolates represented false-positive cultures and that 91 (54%) of the remaining 168 isolates were in 15 DNA fingerprinting clusters, which ranged in size from 2 to 29 isolates. Risk factors for clustering were birth in the United States, African-American ethnicity, homelessness, substance abuse, and male sex. Retrospective epidemiologic analysis of inpatient and outpatient visits by the 91 patients who had clustered isolates revealed only one possible instance of patient-to-patient transmission. CONCLUSIONS: The DNA fingerprinting of all M. tuberculosis isolates from a 1-year period revealed one possible instance of nosocomial transmission and five false-positive M. tuberculosis cultures. However, these results did not lead to changes in infection-control practices or in clinical care. The study findings do not support the use of DNA fingerprinting for nosocomial tuberculosis surveillance, but they suggest that compliance with the CDC tuberculosis infection-control guidelines may control patient-to-patient transmission in high-risk urban hospitals.

Randomized trial of general hospital and residential alternative care for patients with severe and persistent mental illness.

OBJECTIVE: Severe and persistent mental illnesses are often lifelong and characterized by intermittent exacerbations requiring hospitalization. Providing needed care within budgetary constraints to this largely publicly subsidized population requires technologies that reduce costly inpatient episodes. The authors report a prospective randomized trial to test the clinical effectiveness of a model of acute residential alternative treatment for patients with persistent mental illness requiring hospital-level care. METHOD: Patients enrolled in the Montgomery County, Md., public mental health system who experienced an illness exacerbation and were willing to accept voluntary treatment were randomly assigned to the acute psychiatric ward of a general hospital or a community residential alternative. There were no psychopathology-based exclusion criteria. Treatment episode symptom improvement, satisfaction, discharge status, and 6-month pre- and postepisode acute care utilization, psychosocial functioning, and patient satisfaction were assessed. RESULTS: Of 185 patients, 119 (64%) were successfully placed at their assigned treatment site. Case mix data indicated that patients treated in the hospital (N = 50) and the alternative (N = 69) were comparably ill. Treatment episode symptom reduction and patient satisfaction were comparable for the two settings. Nine (13%) of 69 patients randomly assigned to the alternative required transfer to a hospital unit; two (4%) of 50 patients randomly assigned to the hospital could not be stabilized and required transfer to another facility. Psychosocial functioning, satisfaction, and acute care use in the 6 months following admission were comparable for patients treated in the two settings and did not differ significantly from functioning before the acute episode. CONCLUSIONS: Hospitalization is a frequent and high-cost consequence of severe mental illness. For patients who do not require intensive general medical intervention and are willing to accept voluntary treatment, the alternative program model studied provides outcomes comparable to those of hospital care.

Cross-contamination of specimens with Mycobacterium tuberculosis: clinical significance, causes, and prevention.

At the Veterans Affairs Lakeside Medical Center, two episodes of specimen cross-contamination with Mycobacterium tuberculosis were detected during a 54-month period by molecular strain typing using DNA restriction fragment length polymorphism for 3 patients without clinical or radiologic signs of tuberculosis (TB). A cross-contaminated specimen was the only culture-positive specimen for each of the 3 patients. Laboratory features of cross-contamination included acid-fast smear negativity, growth only in broth or solid medium, and growth in solid medium with 5 or fewer colonies. Retrospective analysis demonstrated identical features for occasional culture-positive specimens from 54 patients with TB during the same period. However, productive cough, pleural pain, weight loss, night sweats, chest radiograph results suggestive of TB, positive tuberculin skin testing, and/or multiple culture-positive specimens were invariably present in patients with TB with such specimens. Most patients with TB (50/54; 93%) had multiple specimens positive in culture for M. tuberculosis, and the few patients with TB with single culture-positive specimens were symptomatic. These results indicate that correlation with clinical manifestations is necessary to determine the significance of isolated, acid-fast smear negative, and/or low-yield culture-positive specimens. Although the prevalence of specimen cross-contamination is low (0.1%), possible sources (especially the use of single-reagent delivery systems for multiple specimens) should be eliminated by mycobacteriology laboratories.

Tuberculosis among Tibetan immigrants from India and Nepal in Minnesota, 1992-1995.

OBJECTIVE: To study screening outcomes among a group of Tibetan immigrants at high risk for developing active tuberculosis (TB) after arrival in Minnesota. DESIGN: Retrospective cohort study. PARTICIPANTS: A total of 191 Tibetan immigrants undergoing medical screening. MAIN OUTCOME MEASURES: Occurrence and treatment outcomes of active TB. SETTING: A health maintenance organization and a public TB clinic in Minneapolis, Minn. RESULTS: Positive (induration, > or =10 mm) tuberculin skin test results were documented in 98% of Tibetans, compared with 44% of Vietnamese, 10% of Hmong, and 51% of Russian refugees in Minnesota (P<.001 for each group). Sixteen active cases (8.4%) were confirmed by isolation of Mycobacterium tuberculosis; however, 5 (31%) were culture-negative on initial screening in Minnesota. Seven cases (44%) were diagnosed during initial screening efforts, and 9 cases (56%) were diagnosed a mean of 19 months (range, 10-27 months) after their initial medical evaluation. Of these 9 cases, 6 (38% of all Tibetan cases) had isolates resistant to 1 or more antituberculous drugs, and 3 (19% of all Tibetan cases) were multidrug resistant (MDR TB). All 3 MDR TB cases were culture-negative on initial screening; these cases constituted 75% of the MDR TB isolates in Minnesota in 1994. The presence of MDR TB was associated with a known history of active TB in Asia (P<.02). Any abnormality on chest radiograph noted either during the Immigration and Naturalization Service screening evaluation in India (relative risk [RR], 5.2; P=.006) or on arrival in Minnesota (RR, 6.8; P=.005) was associated with an increased risk of subsequent active TB. CONCLUSIONS: Tuberculosis infection is nearly universal among Tibetans settling in Minnesota. A single screening evaluation failed to detect the majority of TB cases among Tibetans. Even in the face of negative M tuberculosis cultures, persons with a history of active TB require particularly close follow-up.

Residential alternatives to hospitalization for patients with severe and persistent mental illness: should patients with comorbid substance abuse be excluded?

Residential alternatives to hospitalization for adults with severe mental illness in crisis were not designed for, and often exclude, persons with coexisting substance abuse disorders. Given high comorbidity rates, however, it is important to know whether residential alternatives can be effective for patients with dual diagnoses. To explore the impact of comorbidity on treatment outcomes, structured interviews were conducted at admission and discharge with 92 consecutive admissions to a residential alternative. Using the Structured Clinical Interview for DSM-III-R, two groups were identified: 24 patients with and 68 patients without comorbid substance abuse disorders. At admission, the two groups were similar in demographic and clinical characteristics. The treatment was effective independent of comorbidity; at discharge, treatment success, symptom improvement, and patient satisfaction were similar for both groups. Persons with coexisting substance abuse disorders remained in residence a week longer, but the difference was not statistically significant. Residential alternatives appear suitable for patients with dual diagnoses.

Specimen contamination in mycobacteriology laboratory detected by pseudo-outbreak of multidrug-resistant tuberculosis: analysis by routine epidemiology and confirmation by molecular technique.

A cluster of multidrug-resistant tuberculosis sputum isolates led to the detection of specimen contamination in a hospital mycobacteriology laboratory. Thirteen specimens were smear negative but culture positive from one specimen only; 12 appeared to be contaminated. Each of these specimens was processed in the same batch as one or more smear- and culture-positive isolates. Molecular analysis confirmed the traditional epidemiologic, laboratory, and clinical methods of evaluating presumed mycobacterial contamination.

The treatment of acute psychosis without neuroleptics: six-week psychopathology outcome data from The Soteria Project.

BACKGROUND: Today's treatment of acute psychosis usually includes short-term hospitalization and anti-psychotic drug treatment. The Soteria project compared this form of treatment (control) with that of a small, home-like social environment, usually without neuroleptics (experimental). METHOD: Newly diagnosed, young, unmarried persons with DSM-II schizophrenia were randomly assigned to treatment in two experimental and two control settings. Subjects and families were assessed at admission on 29 independent variables. Treatment environments were studied by means of Moos', COPES or WAS scales. Three dependent six week psychopathology outcome measures were collected. RESULTS: The groups were comparable on 25 of 29 admission variables. The environments of the two experimental and two control settings were different from each other. The milieus were similar to each other within each condition. At six weeks, psychopathology in both groups had improved significantly, and similarly, and overall change was the same. CONCLUSION: Specially designed, replicable milieus were able to reduce acute psychotic symptomatology within six weeks, usually without antipsychotic drugs, as effectively as usual hospital ward treatment that included routine neuroleptic drug use.

Suppressive effect of antioxidants on intercellular adhesion molecule-1 (ICAM-1) expression in human epidermal keratinocytes.

Intercellular adhesion molecule-1 (ICAM-1) is strongly expressed by human epidermal keratinocytes during the course of inflammatory skin diseases. To test the possibility that reactive oxygen species produced in the skin during an inflammatory response affect ICAM-1 expression, cultured human epidermal keratinocytes were treated with H2O2 at concentrations that did not damage the cells, and cell-surface ICAM-1 expression was analyzed. Expression of ICAM-1 was induced on keratinocytes by treatment with 300 microM H2O2 for 1 h. The antioxidant N-acetyl-L-cysteine strongly inhibited H2O2-induced ICAM-1 expression, whereas the antioxidants pyrrolidine dithiocarbamate and alpha-tocopherol were less inhibitory. N-acetyl-L-cysteine also suppressed keratinocyte surface expression of ICAM-1 induced by the cytokines interferon-gamma (IFN-gamma) or tumor necrosis factor-alpha (TNF-alpha), whereas pyrrolidine dithiocarbamate and alpha-tocopherol suppressed IFN-gamma-induced surface expression but not TNF-alpha-induced expression. We found that N-acetyl-L-cysteine treatment reduced ICAM-1 mRNA levels when keratinocytes were stimulated with either IFN-gamma or TNF-alpha; however, pyrrolidine dithiocarbamate and alpha-tocopherol had no effect on either IFN-gamma- or TNF-alpha-induced ICAM-1 mRNA levels. Our results indicate that reactive oxygen species may be involved in the skin inflammatory process by increasing epidermal ICAM-1 expression and that some antioxidants may be effective in suppressing the epidermal ICAM-1 expression induced by reactive oxygen species and cytokines in inflammatory skin diseases.

Human aortic endothelial cells express the type I but not the type II receptor for interleukin-1 (IL-1).

Endothelial cells (EC) are very responsive to the proinflammatory cytokine interleukin-1 (IL-1). EC are induced by IL-1 to secrete chemotactic factors and to increase expression of cell surface adhesion molecules leading to increased leukocyte adhesion. Activated EC further contribute to the inflammatory response by secreting additional cytokines. IL-1 interacts with EC through high-affinity cell-surface receptors. However, the low number of receptors present on EC has made characterization difficult. Further, recent evidence has suggested diversity in the responses of EC from different regions of the vascular system. Interested in the effect of IL-1 on early atherosclerotic lesion formation, we have characterized the IL-1 receptors on human aortic endothelial cells (HAEC). Using a direct binding assay, we found that HAEC have 1,000-3,000 IL-1 receptors per cell and bind IL-1 alpha with a Kd of 3.5 x 10(-10) M. We found that a monoclonal antibody specific for the type I receptor completely blocks IL-1 alpha binding. The blocking antibody also completely inhibits the IL-1 induced increase in intracellular adhesion molecule 1 (ICAM-1) expression by HAEC. Using solution hybridization and ribonuclease protection with an antisense probe, a sensitive method for detection of low abundance mRNA species we found that HAEC as well as human umbilical vein EC (HUVEC) have significant levels of mRNA for the type I IL-1 receptor. To test whether HAEC might also contain transcripts for the type II IL-1 receptor, we compared levels of mRNAs by polymerase chain reaction (PCR) amplification of cDNAs reverse-transcribed from total RNA. We found only transcripts for the type I receptor and not the type II receptor in HAEC. Based on this data, we conclude that aortic endothelial cells respond to IL-1 through the type I receptor.

Inhibition of IL-1 beta expression in THP-1 cells by probucol and tocopherol.

The cytokine interleukin-1, IL-1, likely plays an important role in the early stages of atherogenesis. The possible action of probucol and tocopherol on the expression and secretion of IL-1 beta was investigated using the human monocytic leukemia cell line, THP-1. Both probucol and D-alpha-tocopherol inhibit the phorbol ester-induced release of IL-1 beta without altering differentiation. Analysis of IL-1 beta mRNA levels revealed that probucol and tocopherol had an inhibitory effect on the activation of expression of the IL-1 beta gene. The data suggest that the beneficial effects of probucol may be related to inhibition of IL-1 at an early phase of atherosclerotic plaque formation.

Training psychiatrists in the community: a report of the Italian experience.

In 1978 a revolutionary mental health act was passed in Italy. By closing large mental hospitals and replacing them with community programs, it required a radical shift in psychiatric practice. The authors discuss the background philosophy, principles, and practical implications of this change. They describe a model program and training design of a 4-year residency in which psychiatrists learn the skills for community work while actually working in the community. The residency differs from most U.S. residencies in having trainees responsible for patients wherever they are being treated (residents are not rotated between services), its strong team orientation, and the value placed on community work.