Autodisplay of an avidin with biotin-binding activity on the surface of Escherichia coli.

OBJECTIVES: To display a recombinant avidin fused to the autotransporter ShdA to bind biotinylated molecules on the surface of Escherichia coli. RESULTS: Two chimeric protein constructs containing avidin fused to the autotransporter ShdA were expressed on the surface of Escherichia coli DH5α. One fusion protein contained 476 amino acids of the ShdA α and ß domains, whereas the second consisted of a 314 amino acid from α and truncated ß domains. Protein production was verified by SDS-PAGE using an antibody to the molecular FLAG-tag. The surface display of the avidin-shdA fusion protein was confirmed by confocal microscopy and flow cytometry analysis, and the biotin-binding activity was evaluated by fluorescence microscopy and flow cytometry using biotin-4-fluorescein and biotinylated-ovalbumin (OVA). CONCLUSIONS: Expression of a recombinant avidin with biotin-binding activity on the surface of E. coli was achieved using the autotransporter ShdA. This system is an alternative to bind biotinylated molecules to E. coli.

EBV, HCMV, HHV6, and HHV7 screening in bone marrow samples from children with acute lymphoblastic leukemia.

Acute lymphoblastic leukemia (ALL) is the most common cancer in childhood worldwide and Mexico has reported one of the highest incidence rates. An infectious etiology has been suggested and supported by epidemiological evidences; however, the identity of the involved agent(s) is not known. We considered that early transmitted lymphotropic herpes viruses were good candidates, since transforming mechanisms have been described for them and some are already associated with human cancers. In this study we interrogated the direct role of EBV, HCMV, HHV6, and HHV7 human herpes viruses in childhood ALL. Viral genomes were screened in 70 bone marrow samples from ALL patients through standard and a more sensitive nested PCR. Positive samples were detected only by nested PCR indicating a low level of infection. Our result argues that viral genomes were not present in all leukemic cells, and, hence, infection most likely was not part of the initial genetic lesions leading to ALL. The high statistical power of the study suggested that these agents are not involved in the genesis of ALL in Mexican children. Additional analysis showed that detected infections or coinfections were not associated with prognosis.

Trichinella spiralis: intranasal immunization with attenuated Salmonella enterica carrying a gp43 antigen-derived 30mer epitope elicits protection in BALB/c mice.

Trichinellosis is a public health problem and is considered an emergent/re-emergent disease in various countries. The etiological agent of trichinellosis is the nematode Trichinella, which infects domestic animals such as pigs and horses, as well as wild animals and humans. A veterinary vaccine could be an option to control the disease in domestic animals. Although several vaccine candidates have shown promising results, a vaccine against trichinellosis remains unavailable to date. Attenuated Salmonella strains are especially attractive live vectors because they elicit mucosal immunity, which is known to be important for the control of Trichinella spiralis infection at the intestinal level and can be administered by oral or intranasal routes. In this study, the autotransporter ShdA was used to display, on the surface of the Salmonella enterica serovar Typhimurium SL3261, the 210-239 amino acid epitope, (designated as Ag30) derived from the 43 kDa glycoprotein of T. spiralis muscle larvae. The fusion protein elicited antibodies in BALB/c mice that were able to recognize the native epitope on the surface of T. spiralis muscle larvae. Mice immunized by intranasal route with the recombinant Salmonella induced a protective immune response against the T. spiralis challenge, reducing by 61.83% the adult burden at day eight postinfection. This immune response was characterized by the induction of antigen-specific IgG1 and of IL-5 production. This study demonstrates the usefulness of Salmonella as a carrier of nematode epitopes providing a surface display system for intestinal parasite vaccine applications.

Diagnóstico del origen y la causa de la muerte después de la autopsia médico-legal (Parte II) / Diagnosis of the origin and cause of death after the medico-legal autopsy (Part II)

Una de las tareas más difíciles en patología forense puede ser determinar, después de la autopsia, el origen y la causa de la muerte cuando dichos extremos no están claros o son discutibles. Una autopsia técnicamente perfecta es condición necesaria pero no suficiente para cumplir adecuadamente esta decisión. En el presente trabajo se delimitan claramente los conceptos de causa y mecanismo de muerte, muerte natural y muerte violenta. Se revisan muchos de los aspectos relacionados con el diagnóstico del origen y la causa de la muerte; especialmente el enfoque actual del valor de la autopsia en el diagnóstico, la interacción entre traumatismo y enfermedad, la doctrina de la causalidad, la utilización de la CIE-10, la llamada etiología médico-legal, la autopsia psicológica, la "autopsia blanca" o negativa y el origen de la muerte por complicaciones de la terapéutica, etc., todos ellos se ilustran con ejemplos de casos representativos. Se discute el limitado papel del patólogo forense en España en la determinación de la denominada etiología médico-legal. Las conclusiones constituyen un conjunto de recomendaciones para el mejor cumplimiento de este trabajo, entre ellas el abandono de las denominadas causas de muerte inaceptables como parada cardiaca, parada cardiorrespiratoria o muerte cerebral, etc., la utilización siempre que sea posible de los diagnósticos de la CIE-10 y del formato del apartado de la causa de la muerte (parte I y II) del modelo internacional de certificado médico de causa de defunción de la OMS. Todo ello como guía breve que facilite un resultado judicial adecuado y aumente la credibilidad del patólogo forense (AU)

One of the most difficult tasks in forensic pathology is deciding the origin and the cause of death after the autopsy when those issues are unclear or debatable. A technically perfect autopsy is a necessary but not a sufficient condition to adequately fulfil this decision. The present paper clearly defines the concepts of cause and mechanism of death, natural and violent death. We review many aspects related to the diagnosis of the origin and cause of death, especially the current approach of the value of autopsy in the diagnosis, the interaction between trauma and disease, the doctrine of causation, the use of ICD- 10, the so-called manner of death, the psychological autopsy, the negative autopsy and the origin of death from therapeutic complication, and so on., all of which are illustrated with relevant examples. We also discuss the limited role of the forensic pathologist in Spain in determining the so-called manner of death. The conclusions include(s) a list of recommendations for the best performance of this work, including the abandonment of the so-called unacceptable causes of death -as cardiac arrest, cardiopulmonary arrest or brain death, and so on- and whenever possible the adoption of the classification of ICD-10 and the standard paragraph in which the cause of death is informed (part I and II), as proposed by the WHO international certificate of death. These conclusions intend to be a brief guide to provide a fair judicial outcome and enhance the forensic pathologist's credibility (AU)

Diagnóstico del origen y la causa de la muerte después de la autopsia médico-legal (Parte I) / Diagnosis of the origin and cause of death after the medico-legal autopsy (Part I)

Una de las tareas más difíciles en patología forense puede ser determinar, después de la autopsia, el origen y la causa de la muerte cuando dichos extremos no están claros o son discutibles. Una autopsia técnicamente perfecta es condición necesaria pero no suficiente para cumplir adecuadamente esta decisión. En el presente trabajo se delimitan claramente los conceptos de causa y mecanismo de muerte, muerte natural y muerte violenta. Se revisan muchos de los aspectos relacionados con el diagnóstico del origen y la causa de la muerte; especialmente el enfoque actual del valor de la autopsia en el diagnóstico, la interacción entre traumatismo y enfermedad, la doctrina de la causalidad, la utilización de la CIE-10, la llamada etiología médico-legal, la autopsia psicológica, la “autopsia blanca” o negativa y el origen de la muerte por complicaciones de la terapéutica, etc., todos ellos se ilustran con ejemplos de casos representativos. Se discute el limitado papel del patólogo forense en España en la determinación de la denominada etiología médico-legal. Las conclusiones constituyen un conjunto de recomendaciones para el mejor cumplimiento de este trabajo, entre ellas el abandono de las denominadas causas de muerte inaceptables como parada cardiaca, parada cardiorrespiratoria o muerte cerebral, etc., la utilización siempre que sea posible de los diagnósticos de la CIE-10 y del formato del apartado de la causa de la muerte (parte I y II) del modelo internacional de certificado médico de causa de defunción de la OMS. Todo ello como guía breve que facilite un resultado judicial adecuado y aumente la credibilidad del patólogo forense (AU)

One of the most difficult tasks in forensic pathology is deciding the origin and the cause of death after the autopsy when those issues are unclear or debatable. A technically perfect autopsy is a necessary but not a sufficient condition to adequately fulfil this decision. The present paper clearly defines the concepts of cause and mechanism of death, natural and violent death. We review many aspects related to the diagnosis of the origin and cause of death, especially the current approach of the value of autopsy in the diagnosis, the interaction between trauma and disease, the doctrine of causation, the use of ICD- 10, the so-called manner of death, the psychological autopsy, the negative autopsy and the origin of death from therapeutic complication, and so on., all of which are illustrated with relevant examples. We also discuss the limited role of the forensic pathologist in Spain in determining the so-called manner of death. The conclusions include(s) a list of recommendations for the best performance of this work, including the abandonment of the so-called unacceptable causes of death -as cardiac arrest, cardiopulmonary arrest or brain death, and so on- and whenever possible the adoption of the classification of ICD-10 and the standard paragraph in which the cause of death is informed (part I and II), as proposed by the WHO international certificate of death. These conclusions intend to be a brief guide to provide a fair judicial outcome and enhance the forensic pathologist’s credibility (AU)

Two different PABPN1 expanded alleles in a Mexican population with oculopharyngeal muscular dystrophy arising from independent founder effects.

BACKGROUND: Oculopharyngeal muscular dystrophy (OPMD) is a late onset hereditary myopathy of autosomal dominant transmission characterised by ptosis, dysphagia and limb weakness. The disease is caused by short heterozygous expansions of a (GCN)(10) triplet located in the first exon of the PABPN1 gene at chromosome 14q11.1. Most affected individuals from North America and Europe carry a mutant (GCN)(13) allele. Although evidence for a founder mutation effect has been shown in several populations with OPMD, analysis of large groups of patients from different ethnic backgrounds will help to identify the relative contribution of each allele to the disease and a possible genotype-phenotype correlation. METHODS: 22 unrelated patients with OPMD from Mexico, a previously uncharacterised population, were clinically and molecularly analysed. Detailed ophthalmological and clinical examinations were performed in each proband and molecular analysis of the PABPN1 gene was carried out by PCR amplification and allele-specific cloning/sequencing. Two single nucleotide polymorphisms (SNPs) linked to PABPN1 were determined in each individual and in a number of affected first-degree relatives. RESULTS: 15 subjects (68%) carried a mutant (GCN)(15) or (GCG)(11)(GCA)(3)(GCG) PABPN1 allele; the remaining 7 (32%) exhibited an abnormal (GCN)(13) or (GCG)(9)(GCA)(3)(GCG) allele. Analysis of two SNPs linked to PABPN1 strongly suggests that both expanded alleles originate from two independent founder effects. In addition, in this particular population the (GCN)(15) allele was associated with an earlier onset of the disease (mean 46.5 years) compared with the (GCN)(13) allele (mean 54.7 years). CONCLUSION: The results of this study suggest that OPMD in the Mexican population is mostly due to (GCG)(11) or (GCG)(9) PABPN1 expanded alleles arising from two independent founder effect mutations. These findings add to the definition of the genetic features of the disease and to the establishment of a probable genotype-phenotype correlation.

[Asbestos-related diseases in relatives of asbestos exposed workers]. / Patologie asbesto correlate in congiunti di lavoratori del settore cemento-amianto.

Standard asbestos diagnostic protocol was applied to eleven relatives of asbestos exposure ex-workers of a cement factory in Bari. Nine wives and seventeen sons were involved as volunteers in this evaluation. In this group two pleura malignant mesotheliomas (not dose-dependent) two asbestosis and fifteen pleura plaques (dose-dependent) were detected. This situation shows high level of asbestos contamination at home. For all the test patients the contamination most probably occurred because workers carried asbestos substances home from work on their clothes.

[Alcohol related risk survey about health workers]. / Indagine conoscitiva sul rischio alcol correlato nei dipendenti di una azienda ospedaliera.

The link between drinking and consequent injuries during job activities has been stresses by the most important international organizations and by ISTAT. Luckily, the low 125/01 has opened to "occupational doctor" the possibility of analysing the alcohol related risks among particular working classes, i.e. hospital workers. Thus, it has been conducted a survey in our Hospital in order to understand the entity of the alcohol related risk. The 32.59 of the sample was classified as teetotallers, 45.81% was in the range of low risk probability, while the 24.58% was in the high risk one. It has also been noticed a significant link between the highest level of risk and male employees, but nothing in relation with age or the type of job. In conclusion, this study has represented the first step for our next training programs to reduce the alcohol related risks and give a support to those workers who have already drinking problems.

[Pleural mesothelioma in barman with probable occupational exposure to asbestos]. / Mesotelioma pleurico in barista con probabile esposizione professionale ad amianto.

The case of a barman who suffer from Malignant Mesothelioma (MM) has been signaled. For this case it has been documented a possible source of occupational exposure to asbestos caused by the presence of asbestos components in the professional espressos coffee machines. In some gaskets which are part of these coffee machines, we verified the presence of chrysotile fibers. Italian National Mesothelioma Register have reported a MM case with a professional origin arised in a barman with a certain diagnosis in 1999 (Tuscany Region).

[Health surveillance of subjects formerly exposed to asbestos in Puglia]. / Sorveglianza sanitaria degli ex-esposti ad amianto in Puglia.

The massive use of asbestos affected several industrial areas of Apulian region in the last decade. It has been estimated that in our country there are thousands of asbestos exposed workers and about 10.000 of them received CONTARP-INAIL document attesting their asbestos exposure for over ten years ex D.L. 257/92. Owing to the lack of any regional rules and guidelines on the medical surveillance of asbestos ex-exposed people, since 1994 our operative hospital unit has activated a sanitary protocol in order to satisfy growing requests of assistance and emergent controls for this part of people. During a 12 year activity 629 asbestos ex-exposed workers have been examined. 21% of these workers had two checks, thanks to these the recognition of 22 new tumoral and non-tumoral asbestos-correlated pathologies has been possible. This experience makes it clear that the activation of new procedures for early diagnoses of some asbestos-correlated pathologies is necessary, in fact more proposals for a redevelopment of sanitary assistance in the regional field are suggested.

Risk factors for childhood burn injuries: a case-control study from Greece.

During a 12-month period 239 children who presented with a burn injury at the Emergency Department of a teaching children's hospital in Athens, with city-wide coverage, and 239 gender- and age-matched controls with minor non-injury ailments were interviewed. The questionnaire covered sociodemographic characteristics of the children and their families, information allowing the construction of a burn avoidance index in their homes and items from the Achenback scale that were synthesized into a child activity score. The data were analyzed through conditional logistic regression. In general, socio-demographic variables were not of overwhelming importance, although some of the findings indicate that supervision lapses and barefoot walking of gypsy children increase the risk of burn injuries. The kitchen in an inherently high risk place for injuries and the powerful inverse association of the burn avoidance index with burn injury risk points towards steps that could be easily taken and impart substantial protection. There was no evidence in this study of burn injury proneness or that hyperactivity of the child increased the risk of burn injury; indeed, the results point in the opposite direction. Our results strongly support the view that childhood burn injuries are largely environmentally conditioned and, accordingly, easily preventable.

Diet during pregnancy in relation to birthweight in healthy singletons.

We have investigated the relationship between consumption of food groups and intake of energy-generating macronutrients on the one hand, and birthweight on the other among apparently healthy singleton, term babies. Three hundred and sixty-eight women who delivered in six maternity clinics in two Greek cities during specified days over an 8-month period completed a 190-item, interviewer-administered, validated, semiquantitative food frequency questionnaire. Study participants also provided information on sociodemographic, reproductive and lifestyle variables. Data were analysed using multiple regression modelling. Nutritional variables were energy-adjusted, and non-nutritional correlates of birthweight were accounted for. The analysis revealed most of the established non-nutritional associations of birthweight -- an indication of study validity. Among food groups, meat and meat products and fish and sea food were suggestively associated with increased birthweight (two tailed P-values 0.08 and 0.16, respectively). Among energy-generating nutrients, monounsaturated fat was positively associated with birthweight and significantly so in several of the models. We consider our findings are considered as compatible with hypotheses linking fish and meat intake to fetal growth and as indicative of a positive association between intake of monounsaturated fat and birthweight.

Aplastic anemia in a young coke plant worker.

The objective of this study was to make a contribution to the debate on the cause-effect relationship between low-dose exposure to benzene and onset of hemopathies. We report the case of a coke plant worker suffering from aplastic anemia. Before being hired at the coke plant, he underwent a medical examination including a blood cell count: no disease or abnormalities of the blood crasis were found. For 3 years the patient was then exposed to gas containing-as measured in environmental investigation carried out at the coke plant-concentrations of benzene lower than TLV-TWA ACGIH (measured values 21-109 micrograms/m3). In the absence from the patient's history of any exposure to other myelotoxic agents and of any earlier pathology causing aplastic anemia, we assume there is a relationship between exposure to low levels of benzene and onset of the disease. However, it is very important to consider that exposure to low levels of benzene could promote myelotoxic reactions when the working environment contains other substances that may act synergistically or compete for the same metabolism sites, or when carcinogenic substances are present in the working environment.

Cimetidine prevents and partially reverses CCl4-induced liver cirrhosis.

Liver injury produced by CCl4 depends on its metabolism by the liver cytochrome P450 enzyme system to a highly reactive intermediate (CCl3.). Cimetidine impairs cytochrome P450 and stimulates regenerative processes acting on DNA synthesis. This work was performed to investigate whether cimetidine may prevent CCl4-induced liver cirrhosis. Male Wistar rats were used: animals in group 1 received CCl4 (0.04 g per 100 g, i.p.) three times a week for 8 weeks; group 2 was treated with CCl4 plus cimetidine (120 mg kg-1, p.o.) three times a week for 8 weeks; group 3 received CCl4 for 8 weeks and then cimetidine for 4 weeks. Alkaline phosphatase, gamma-glutamyl transpeptidase (gamma-GTP) and alanine aminotransferase (ALT) activities, as well as protein and bilirubin, were measured in serum; collagen and lipoperoxidation were quantified in liver. Intoxication with CCl4 increased (P < 0.05) serum activities of alkaline phosphatase, gamma-GTP and ALT, and bilirubin concentration; liver collagen and lipoperoxidation were also increased. Cimetidine treatment prevented or reverted the increases in the three enzyme activities and in bilirubin content and the fall in proteins. It is worth noting that cimetidine co-treatment completely prevented both the increase in collagen content and the lipid peroxidation. The protective effect of cimetidine can be attributed to a reduction in cytochrome P450. However, it could also stimulate regenerative processes.