Modulation of human endogenous retroviruses and cytokines expression in peripheral blood mononuclear cells from autistic children and their parents.

BACKGROUND: Putative pathogenic effects mediated by human endogenous retroviruses (HERVs) in neurological and psychiatric disorders in humans have been extensively described. HERVs may alter the development of the brain by means of several mechanisms, including modulation of gene expression, alteration of DNA stability, and activation of immune system. We recently demonstrated that autistic children and their mothers share high expression levels of some HERVs and cytokines in peripheral blood mononuclear cells (PBMCs) ex vivo, suggesting a close mother-child association in Autism Spectrum Disorder (ASD). RESULTS: In the present study, PBMCs from autistic children and their parents were exposed to stimulating factors (Interleukin-2/Phytohaemagglutinin) or drugs, as Valproic acid and Efavirenz. The results show that HERVs and cytokines expression can be modulated in vitro by different stimuli in PBMCs from autistic children and their mothers, while no significant changes were found in PBMCs ASD fathers or in controls individuals. In particular, in vitro exposure to interleukin-2/Phytohaemagglutinin or valproic acid induces the expression of several HERVs and cytokines while Efavirenz inhibits them. CONCLUSION: Herein we show that autistic children and their mothers share an intrinsic responsiveness to in vitro microenvironmental changes in expressing HERVs and pro-inflammatory cytokines. Remarkably, the antiretroviral drug Efavirenz restores the expression of specific HERV families to values similar to those of the controls, also reducing the expression of proinflammatory cytokines but keeping the regulatory ones high. Our findings open new perspectives to study the role of HERVs in the biological mechanisms underlying Autism.

Formaldehyde exposure and irritative effects on medical examiners, pathologic anatomy post-graduate students and technicians.

BACKGROUND: Exposure to formaldehyde (FA) causes irritative effects and induces nasopharyngeal cancer; the International Agency for Research on Cancer (IARC, Lyon) classified FA as carcinogenic to humans, Group 1. Many studies have been published so far concerning the occupational exposure of industrial workers, embalmers, pathologists and anatomists to FA but very few data regarding medical examiners are available. METHODS: To assess the extent to which subjects were exposed to FA, airborne concentrations of this chemical were measured by High Performance Liquid Chromatography (HPLC). In September-October 2006 we examined the personnel, which worked in an autopsy room (medical examiners) and in three laboratories of pathologic anatomy of the University Medical School of Bari, Policlinico Hospital, Southern Italy. Irritative effects were also investigated. RESULTS: All the personal exposure data obtained exceeded the National Institute for Occupational Safety and Health Threshold Limit Value-Time Weighted Average (NIOSH TLV-TWA: 0.02 mg/m(3)) and, in a few cases, even the American Conference of Industrial Hygienists Threshold Limit Value-Ceiling level (ACGIH TLV-C: 0.37 mg/m(3)). CONCLUSION: Irritative effects in more than 50% of the workers enrolled, increasing the risk of injuries.

[Fitness to work and diabetes in health care workers]. / Idoneità lavorativa e diabete in operatori sanitari.

Diabetes mellitus is a disease of great social relief that has a deep impact also in working life. Our short paper wants to highlight some critical aspects that occupational physician must deal with both in preventive and periodic health evaluation for diabetic subjects. In our study, performed on health care workers, we reported 16 cases of diabetes; 10 of them were nurses, 5 sanitary auxiliaries and 1 sanitary technical operator. The judgment has been of full suitability to the specific task for 7 subjects, whereas for 4 workers it has been stated suitability with limitation for problems related with diabetes or its complications. The judgment with limitation for the other 5 subjects was due to pathologies not related with diabetes. The shift work, and mainly the night shift, is the risk that diabetic subjects cannot effort when the disease is not sufficiently compensated. The small number of diabetic workers, 16 over 1994 visited (0.8%), suggests for a spontaneous choice of diabetic people towards other kind of work without shift risk.

[Application and effects of smoking ban in bars and restaurants of Rome]. / Applicazione ed effetti del divieto di fumare (Legge n. 3 del 16/01/2003) nei bar e ristoranti di Roma.

Both active and passive tobacco smoke is carcinogenic. In the last years the most important countries of European Community developed and passed smoke-free public places and smoke-free workplace legislations. The aim of this study was to investigate the real application of smoking ban in bars and restaurants of Rome and to value social, economic and health effects caused by the application of the law. The study was carried out in 200 public places (100 restaurants and 100 bars) with an inspection of the sites and the administration of a questionnaire to the managers of the public places. Results demonstrate that smoking ban in public places is widely respected and that the application of the law had a very positive impact on the quality of life and health of workers and general population.

[Health surveillance in health care workers]. / La sorveglianza sanitaria degli operatori sanitari: esperienza in un Policlinico universitario.

In a complex reality as the hospital, occupational physician plays a key role in risk prevention through health surveillance. The medical examination is intended to highlight any possible deterioration of workers' health and to detect workers' hypersusceptibility to occupational hazards. We report biostatistic data resulted from health surveillance conducted on health care workers in 2005 and 2006 in a universitary hospital, with particular regard to the judgement of the fitness to work and the reasons that has determined it. Our report, in agreement with data available in literature, shows that manual patient lifting is one of the most common professional hazards within the hospital and occupational physician must pay a special attention to it, promoting an integrated answer.

[Prevalence and risk factors of urinary incontinence in female workers of hotels]. / Prevalenza e fattori di rischio dell'incontinenza urinaria nelle lavoratrici del settore alberghiero.

The International Continence Society defines urinary incontinence (UI) as "a condition in which involuntary loss of urine is a social or hygienic problem and is objectively demonstrable". There are three different jorms of UI. stress urinary incontinence, urge urinary incontinence and mixed incontinence. The aim of this study was to investigate the prevalence of UI in a group of female workers in the hotel sector. The International Consultation on Incontinence Questionnaire Urinary Incontinence short form (ICIQ-UI Short Form) was administered to all female workers and data were collected about age, body mass index, number of vaginal and Caesarean delivery. Results showed a prevalence of UI widely bigger in the plans waitress than in video display terminal workers and suggest the hypothesis that manual handling of loads representing a possible occupational risk for UI.

[Latex gloves dermatitis in health care workers]. / Esperienza di dermatiti da guanti in lattice nel personale sanitario di un Policlinico Universitario.

With regard to health care workers the irritative contact dermatitis represents about the 80% of all the dermopathies in sanitary staff whereas the allergic contact dermatitis covers approximately the 20% of the professional dermatoses. In our study 4 cases of allergy to latex in hospital nurses are presented; the clinical history is described for each of them as well as the resulting judgment of suitability to the specific work. In general population the allergy to latex is estimated to be approximately 1-6%; in sanitary staff it rises to 5-12%. We have to observe that not all the sensitive subjects show symptoms of allergy. Actually the 4 cases described represent less than 1% of the surveyed group. The sensitization is likely to be so reduced also thanks to the application of preventive guide-lines such as the one proposed by NIOSH. In Italy the criteria in preventing allergic reactions to latex are illustrated in a consensus document issued by a study-team from Italian Association of the Health Workers.

Adenosine A2A receptor dysfunction correlates with age at onset anticipation in blood platelets of subjects with Huntington's disease.

Huntington's disease (HD) may manifest at an earlier age in affected offspring than in transmitting parents. Earlier onset in successive generations (anticipation) only partially depends on intergenerational parent-child elongation of the CAG expanded mutation. An aberrant amplification of adenosine A(2A) receptor signaling documented in peripheral blood cells of subjects with HD implies that this cellular dysfunction may be related to clinical and genetic features. Prompted by evidence of higher receptor densities in siblings of HD subjects with stronger onset anticipation, in this study we investigated a possible relationship between A(2A) receptor densities and age at onset. We measured adenosine A(2A) receptor densities in blood cell platelets from 32 patients with HD and healthy control siblings, and sought a possible linear correlation between maximum platelet A(2A) receptor binding (B(max)) values for the whole cohort of HD subjects and anticipation in years. The increased B(max) values for the 32 subjects with HD (220 in patients vs. 137 in healthy control subjects, P = 0.0001) correlated significantly with anticipation in years (r2, 0.48, P = 0.0001 by linear correlation analysis). An increased platelet A(2A) receptor B(max) may belong in a cascade of toxic events leading to earlier onset of HD: as such it could be a useful marker of onset anticipation.

Italian Huntington disease patients--data and tissue bank.

We have collected clinical and genetic data on Huntington disease (HD) patients and their families over the last 5 years at the Unit of Neurogenetics, IRCCS Neuromed of Pozzilli (IS), Italy. Data on 854 mutation carriers are included in the data bank, together with a large number of DNA samples, blood, and other tissues. In particular, lymphoblastoid cell lines from 100 patients, including subjects carrying very rare genetic conditions (CAG mutation homozygosity, juvenile and infantile onset, pre-mutations) have been established. For all these initiatives ethical approval from the bioethics committee was obtained. We wish to extend this initiative to all families, investigators, and institutions within and, possibly outside, the Italian border in an attempt to enlarge the bank and to institute a HD Research Roster.

Reversed-phase high-performance liquid chromatography versus spectrophotometric assay for thimerosal in Cuban recombinant hepatitis B vaccine.

A reversed-phase liquid chromatographic method was applied to study the stability of thimerosal in Cuban recombinant hepatitis B vaccine samples stored under different temperature conditions. Salicylic acid was used as internal standard, it allowed one to determine the thimerosal in the presence of its degradation products. Good stability of the preservative was demonstrated in vaccine samples for as long as 6 years. The same results were obtained when the vaccine samples were incubated at 37 and 45 degrees C during 30 days. The results were in compliance with the microbiological test for determining the effectiveness of antimicrobial preservative in these samples.

Distorted segregation resulting from pea chromosome reconstructions with alien segments from Pisum fulvum.

Pea (Pisum sativum L.) satellited chromosome reconstructions were analyzed by cytologic markers to identify segregation distortion events. The presence of modified chromosomes was evaluated on the basis of additional rDNA genes, an extra and a longer satellite, all derived from chromosome 5 and chromosome 7 from P. fulvum Sibth. & Sm. The segregation of modified satellited chromosome 5 was monitored through fluorescent in situ hybridization with rDNA probe; it fitted the expected 1:2:1 ratio after self-pollination of a heterozygous genotype for modified chromosome 5. In different genotypes, which were heterozygous for both modified chromosomes 5 and 7, the combined segregation of these chromosomes showed the occurrence of seven karyotype classes instead of the expected nine. The classes with modified chromosome 7 and without modified chromosome 5, whether heterozygous or homozygous, were absent. The hypothesis of gamete selection was rejected since the expected segregation ratio of 5:3:1 was significant by chi-square test. Based on the other hypothesis of postzygotic selection, the segregation ratio did not show a significant deviation from the expected 9:3:1 ratio, thereby indicating that embryo abortion caused the segregation distortion (SD). The hypothesis of the SD system involving two loci carried by the alien satellites of modified chromosomes 5 and 7 is discussed in relation to the evolution of the P. fulvum genome.

The coxsackie-adenovirus receptor (CAR) is used by reference strains and clinical isolates representing all six serotypes of coxsackievirus group B and by swine vesicular disease virus.

Group B coxsackieviruses are etiologically linked to many human diseases, and cell surface receptors are postulated to play an important role in mediating their pathogenesis. The coxsackievirus adenovirus receptor (CAR) has been shown to function as a receptor for selected strains of coxsackievirus group B (CVB) serotypes 3, 4, and 5 and is postulated to serve as a receptor for all six serotypes. In this study, we demonstrate that CAR can serve as a receptor for laboratory reference strains and clinical isolates of all six CVB serotypes. Infection of CHO cells expressing human CAR results in a 1000-fold increase in CVB progeny virus titer compared to mock transfected cells. CAR was shown to be a functional receptor for swine vesicular disease virus (SVDV), as CHO-CAR cells but not CHO mock transfected controls were susceptible to SVDV infection, produced progeny SVDV, and developed cytopathic effects. Moreover, SVDV infection could be specifically blocked by monoclonal antibody to CAR (RmcB). SVDV infection of HeLa cells was also inhibited by an anti-CD55 MAb, suggesting that this virus, like some CVB, may interact with CD55 (decay accelerating factor) in addition to CAR. Finally, pretreatment of CVB or SVDV with soluble CAR effectively blocks virus infection of HeLa cell monolayers.

The tyrosine kinase p56lck is essential in coxsackievirus B3-mediated heart disease.

Infections are thought to be important in the pathogenesis of many heart diseases. Coxsackievirus B3 (CVB3) has been linked to chronic dilated cardiomyopathy, a common cause of progressive heart disease, heart failure and sudden death. We show here that the sarcoma (Src) family kinase Lck (p56lck) is required for efficient CVB3 replication in T-cell lines and for viral replication and persistence in vivo. Whereas infection of wild-type mice with human pathogenic CVB3 caused acute and very severe myocarditis, meningitis, hepatitis, pancreatitis and dilated cardiomyopathy, mice lacking the p56lck gene were completely protected from CVB3-induced acute pathogenicity and chronic heart disease. These data identify a previously unknown function of Src family kinases and indicate that p56lck is the essential host factor that controls the replication and pathogenicity of CVB3.

The complete consensus sequence of coxsackievirus B6 and generation of infectious clones by long RT-PCR.

The full length sequence for the human pathogen coxsackievirus B6 (CVB6, Schmitt strain) has been determined. We used long RT-PCR to generate full length DNA amplicon of CVB6, and then directly sequenced the amplicons. One-step cloning of the full length amplicon enabled us to obtain an infectious clone of CVB6. RNA generated from CVB6 amplicon DNA or CVB6 clones, by transcription with T7 RNA polymerase, was demonstrated to be infectious upon transfection into HeLa cells in vitro. The CVB6 genome is characteristic of enteroviruses, with a 5'-non-translated region (743 nucleotides) followed by an open reading frame (encoding a 2184 amino acid polyprotein) and a 3'-non-translated region (100 nucleotides) and polyadenylated tail. The predicted amino acid sequence of CVB6 clustered with the other CVB serotypes and swine vesicular disease virus (SVDV).

Cardiovirulent coxsackieviruses and the decay-accelerating factor (CD55) receptor.

Group B coxsackieviruses are etiologically linked with many human diseases including acute myocarditis and associated chronic dilated cardiomyopathy. Well-established CVB3 cardiovirulent strains (CVB3c(s)) with known phenotypic difference have been used to study the pathogenesis of virus-induced heart disease. The receptor-binding characteristics of cardiovirulent CVB3 are not known, but may represent one mechanism accounting for differences in disease virulence. In this study, interactions between CVB3c(s) and the decay-accelerating factor (DAF or CD55) cell surface receptor were examined. Anti-DAF monoclonal antibodies (MAbs) blocked virus binding and infection of susceptible HeLa cells. Virus binding was significantly reduced by treatment of these cells with phosphatidylinositol phospholipase C enzyme, which rendered them DAF-deficient CVB3c(s) exhibited a differential propensity for the DAF receptor, as several cardiovirulent strains interacted more strongly than others. However, virus binding and infection was always most effectively blocked by MAbs directed against the SCR 2 and 3 domains of DAF, suggesting that binding occurs at a similar site(s) on the molecule for all strains. Virus binding and internalization were associated with DAF down-regulation at the cell surface, as monitored by flow cytometry analysis. Cardiovirulent CVB3 did not interact with molecules functionally and/or structurally related to DAF, including CD35, CD46, Factor H, or C4-binding protein. Adenovirus type 2 (Ad2) does not use the DAF receptor. However, competitive binding assays between Ad2 and CVB1-6, CVB3c(s), anti-DAF MAbs, or DAF-reduced cells indicated that DAF is associated with Ad2 receptors on the HeLa cell membrane. In summary, this study indicates that DAF is an attachment receptor for cardiovirulent CVB3 and that DAF interaction may be important in the pathogenesis of CVB-mediated heart disease.

Identification of bacteria in water for pharmaceutical use.

Different systems for the obtention of water used in Biopharmaceutical Industry were characterized from the bacteriological point of view. Determination of aerobic mesophilic microorganisms was performed; as well as the isolation of contaminant microorganisms for what the techniques of membrane filtration was used. For the identification of the more representative species there were made conventional biochemical tests and quick systems: API. The results show that water serving as tap water for purification systems fulfill with the microbiological requirements to this kind of water. All the isolated microorganisms were gram-negative bacteria characteristics of this environment: Pseudomonas putida, Xanthomonas maltophilia, Aeromonas salmonicida ssp. salmonicida, Flavimonas coryzihabitans and Acinetobacter iwoffi. The ultrafiltration and distillation tested systems fulfill with the established microbiological limits, except for deionization and distilled water storing systems. The isolation showed that approximately the 76.9% were of Gram-negative bacteria, the 14.6% of Gram-positive cocci and the 8.5% Gram-positive sporulated bacilli. The most representative genus of purified water were: Pseudomonas, with the higher percent of incidence, Staphylococcus, Bacillus, Flavobacterium, Sphingomonas, Aeromonas and Agrobacterium.

High levels of cerebrospinal fluid IgM binding to myelin basic protein are associated with early benign course in multiple sclerosis.

We assessed human myelin basic protein (MBP) binding IgM levels in CSF. MBP is the most studied putative antigen in multiple sclerosis (MS) and immune responses against it may be involved in the demyelination process. We also correlated these levels with EDSS score and other parameters of disease progression and prognosis, both at the time of CSF analysis and during follow-up. CSF IgM anti-MBP levels were assayed by measuring total IgM levels with solid-phase ELISA in CSF samples from 66 patients with relapsing-remitting MS, 11 subjects without neurological diseases, 20 patients with non-inflammatory neurological diseases and 7 patients with lymphocytic meningitis, before and after immunoabsorption with human MBP. Confirmation of IgM binding specificity was performed by immunoblotting of positive CSF samples onto MBP coated-nitrocellulose sheets. Clinical evaluation (disability score, number and time of attacks) was performed during a mean follow-up of 2.7 +/- 1.1 years. 23 of the 66 relapsing-remitting MS patients (33.8%) had elevated IgM anti-MBP levels. In this patient subgroup, IgM anti-MBP levels correlated with the IgM index (r = 0.71; P = 0.0001), but not with CSF/serum albumin (r = 0.08; P = 0.72). In the first year of follow-up, patients with low IgM anti-MBP suffered from more numerous attacks than those with elevated levels (0.86 +/- 0.63 versus 0.43 +/- 0.58; P = 0.017). Patients with high IgM binding to MBP had a first attack during follow-up in a significantly higher time than those with low binding (28.87 +/- 4.7 versus 17 +/- 2.6 months, respectively; P = 0.005) and reached a decrease of 0.5 EDSS point significantly faster than those with low IgM (16.17 +/- 1.2 versus 29.7 +/- 2.6 months, respectively; P = 0.0002). A similar significant finding was observed when the time to reach low disability score (EDSS < or = 2.0) was analyzed (10.7 +/- 2.57 +/- 3.3 months, respectively; P = 0.014). These findings demonstrate that in a subgroup of MS patients, elevated CSF levels of IgM anti-MBP are associated with early favorable course and therefore suggest that IgM binding to MBP could be a possible prognostic marker in relapsing-remitting MS to select early MS patients for future trials.

Viral myocarditis: balance between viral infection and immune response.

Myocarditis is an inflammatory disorder of the heart muscle, and is often underdiagnosed in clinical practice. It presents more dramatically in the young with acute heart failure and more insidiously in adults with chronic dilated cardiomyopathy. The etiology is most often viral in North America, with a wide spectrum of natural history. The majority of patients recover spontaneously, but those with persistent ventricular dysfunction face a 20% one-year mortality. Myocarditis initiates as viral disease, and molecular techniques have confirmed viral persistence. The immune response follows as a two-edged sword-both inadequate and excessive immune responses lead to disease. Finally, the myocyte is the target of the above processes, and expresses molecular, cytokine and vascular changes that lead to dilated cardiomyopathy and heart failure. The gold standard for diagnosis still relies on the overly strict Dallas criteria for evaluating myocardial biopsies. Molecular techniques are playing an increasingly important role in both diagnosis and prognosis. Clinical suspicion is still the key towards an early diagnosis. Treatment must be early and persistent-generally supportive initially, with immunosuppression now playing a secondary role in temporizing those with exuberant immune response. Newer treatments for dilated cardiomyopathy such as amlodipine and carvedilol are equally appropriate for postmyocarditis patients. Future treatment may involve specific biological agents, immune therapy, antiviral strategies and molecular gene therapy.