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.

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.

Favorable left ventricular remodeling following large myocardial infarction by exercise training. Effect on ventricular morphology and gene expression.

Continued adverse remodeling of myocardium after infarction may lead to progressive ventricular dilation and heart failure. We tested the hypothesis that exercise training in a healed myocardial infarction-dysfunction rat model can favorably modify the adverse effects of ventricular remodeling including attenuation of abnormal myosin gene expression. Sprague-Dawley rats were subjected to either proximal LAD ligation or sham operation. At 5 wk after the operation, animals were randomly assigned to sedentary conditions or 6 wk of graduated swim training, creating four experimental groups: infarct sedentary (IS), infarct exercise (IE), sham sedentary (SS), and sham exercise (SE). At 11 wk all rats were sacrificed and analyzed. Compared to sedentary infarct controls, exercise training attenuated left ventricular (LV) dilation and allowed more hypertrophy of the non infarct wall. The exercise-trained hearts also showed a reduction in the estimated peak wall tension. Northern blot analysis showed an increase in beta-myosin heavy chain expression in the hearts of the sedentary infarction group soon after infarction when compared to sham controls. However, with exercise training, there was a significant attenuation of the beta-myosin heavy chain expression in the myocardium. Exercise training in a model of left ventricular dysfunction after healed myocardial infarction can improve the adverse remodeling process by attenuating ventricular dilation and reducing wall tension. The abnormal beta-myosin expression was also attenuated in the exercise trained group. This is evidence that abnormal gene expression following severe myocardial infarction dysfunction can be favorably modified by an intervention.

[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.

[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.

[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.

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.

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).

Effects of interparental violence on the psychological adjustment and competencies of young children.

Preschool children (N = 107) were divided into 4 groups on the basis of maternal report; home and shelter groups exposed to verbal and physical conflict, a home group exposed to verbal conflict only, and a home control group. Parental ratings of behavior problems and competencies and children's self-report data were collected. Results show that verbal conflict only was associated with a moderate level of conduct problems: verbal plus physical conflict was associated with clinical levels of conduct problems and moderate levels of emotional problems; and verbal plus physical conflict plus shelter residence was associated with clinical levels of conduct problems, higher level of emotional problems, and lower levels of social functioning and perceived maternal acceptance. Findings suggests a direct relationship between the nature of the conflict and residence and type and extent of adjustment problems.

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.

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.

Neoadjuvant chemotherapy with cisplatin, epirubicin and VP-16 for stage IIIA-IIIB non-small-cell lung cancer: a pilot study.

Twenty patients with stage IIIA-IIIB non-small-cell lung cancer were treated with cisplatin, epirubicin and VP-16 (PEV) neoadjuvant chemotherapy (CDDP, 70 mg/m2, i.v., d 1; EDX, 60 mg/m2, i.v., d 1; VP-16, 100 mg/m2, i.v., d 1-2-3; every 3 weeks). A partial response was obtained in 11 cases (55%), stable disease in 3 cases (15%), and progressive disease in 6 cases (30%). After chemotherapy, 8 (40%) patients, all achieving a partial response, were elegible for surgery: 5 (25%) had a complete resection (4 IIIA and 1 IIIB) and 3 (15%) an incomplete resection. The treatment was well tolerated. These data show that PEV is an active regimen for neoadjuvant chemotherapy in NSCLC and recommend this therapeutic approach for stage IIIA patients.

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.

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.

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.

Viral infection and the pathogenesis of dilated cardiomyopathy.

Long-term follow-up studies of patients with suspected viral myocarditis reveal progression to dilated cardiomyopathy (DCM) in a significant number of cases. Thus, an underlying viral etiology has been hypothesized in the pathogenesis of ongoing heart disease that leads to DCM. Recent application of molecular biology in clinical diagnosis has strengthened this hypothesis. By use of probe hybridization and polymerase chain reaction, enteroviral RNA has been detected in the myocardium of patients at all stages of the disease process: myocarditis, chronic heart disease, and DCM. Experimental murine models of enterovirus-induced heart disease provide a framework for examining the pathogenic mechanisms. Viral cytotoxicity, immunological responses, viral RNA persistence, and spasm of the coronary microvasculature are all implicated in the ongoing disease process. Abnormal cardiac function and heart failure are attributed to the pathological changes that occur.

Quantitation of enteroviral RNA by competitive polymerase chain reaction.

The polymerase chain reaction (PCR) is a new diagnostic technique for the detection of enteroviral infection; however, it currently provides only qualitative results. The aim of this study was to adapt PCR for the accurate quantitation of enteroviral RNA in clinical specimens. For this purpose, we designed a standard RNA which was homologous to sequences at the 5' end of the coxsackie B3 enterovirus genome but contained a single-base-pair mutation which created a novel internal restriction site. Serial dilutions of this standard template RNA were mixed with a fixed concentration of coxsackie B3 enterovirus RNA. The viral and standard templates were reversed transcribed to cDNA and coamplified by PCR, and a comparison of the radioactive PCR products was made. Since the templates were both present in a single reaction tube and competed for the same primers, the ratio of products remained proportional throughout the amplification process. By this approach, a fourfold-difference in viral titer was clearly distinguishable. Moreover, we were able to accurately quantitate as few as 15 50% tissue culture infectious doses, which reflects common clinical viral titers. This study lays the foundation for quantitation of enteroviral RNA in clinical specimens and establishes a technique that can readily be applied to the diagnosis of enteroviral infection.