First identification of Mycobacterium avium paratuberculosis sheep strain in Argentina

We here identified for the first time the presence of Mycobacterium avium paratuberculosis (MAP) sheep (S) strain in Argentina. IS900 polymerase chain reaction (PCR) was positive. The S strain was compared with MAP cattle (C) strains by using IS1311 PCR-restriction endonuclease analysis (PCR-REA), multiplex PCR and restriction fragment length polymorphism (RFLP) analysis.

Predictores de coqueluche al ingreso en lactantes hospitalizados con infección respiratoria aguda baja / [Pertussis predictors in hospitalized infants with acute lower respiratory tract infection].

BACKGROUND: Pertussis, or whooping cough, caused by Bordetella pertussis (BP) is a re-emerging problem in our environment. Although generally considered that the disease is relatively easy to identify infections respiratory syncytial virus (RSV) can present with similar symptoms in infants remains difficult discrimination. OBJECTIVE: Compare clinical symptoms at admission and complementary studies in infants hospitalized with acute respiratory infection (ARI) and RSV by BP to establish markers that enable their early clinical prediction. MATERIALS AND METHODS: Observational, analytical, case-crossover cross comparing younger than 6 months hospitalized with suspected IRA and pertussis (2007-2012) in which BP identified (PCR and culture) and / or VRS (immunofluorescence in nasal secretions). Coinfections were excluded. Bivariate analysis was performed by calculating OR with 95

CI. Were considered significant at p <0.05. The variables studied were age, sex, hits cough, cyanosis, vomiting, apnea, wheezing and CBC with differential RESULTS: We included 174 infants, 72 (41

) BP and 102 (59

) VRS. Age 2 ± 1 months (range :1-6). In both groups was documented cough and wheeze (OR: 1.2 (0.9 to 1.5) p: 0.1 and OR = 0.9 (0.8 to 1.06) p: 0.2, respectively ). Cyanosis (87

, OR: 13.4 p <0.01) and vomiting (26

, OR: 3.4 p <0.01) were more frequent in infants with BP. The absolute lymphocyte count was significantly higher in children with BP (9387 ± 6317 vs. 5127 ± 2766, p <0.01). By ROC curve was identified at 9000 cells / ml as the best point to differentiate VSR BP (AUC = 0.73, 95

CI :0,64-0, 81). CONCLUSIONS: In infants under 6 months with IRA income presence of apnea, cyanosis and lymphocytosis allowing predict significantly differentiate between pertussis those with RSV infections.

Can course format influence the performance of students in an advanced cardiac life support (ACLS) program?

Advanced cardiac life support (ACLS) is a problem-based course that employs simulation techniques to teach the standard management techniques of cardiovascular emergencies. Its structure is periodically revised according to new versions of the American Heart Association guidelines. Since it was introduced in Brazil in 1996, the ACLS has been through two conceptual and structural changes. Detailed documented reports on the effect of these changes on student performance are limited. The objective of the present study was to evaluate the effect of conceptual and structural changes of the course on student ACLS performance at a Brazilian training center. This was a retrospective study of 3266 students divided into two groups according to the teaching model: Model 1 (N = 1181; 1999-2003) and Model 2 (N = 2085; 2003-2007). Model 2 increased practical skill activities to 75 percent of the total versus 60 percent in Model 1. Furthermore, the teaching material provided to the students before the course was more objective than that used for Model 1. Scores greater than 85 percent in the theoretical evaluation and approval in the evaluation of practice by the instructor were considered to be a positive outcome. Multiple logistic regression was used to adjust for potential confounders (specialty, residency, study time, opportunity to enhance practical skills during the course and location where the course was given). Compared to Model 1, Model 2 presented odds ratios (OR) indicating better performance in the theoretical (OR = 1.34; 95 percentCI = 1.10-1.64), practical (OR = 1.19; 95 percentCI = 0.90-1.57), and combined (OR = 1.38; 95 percentCI = 1.13-1.68) outcomes. Increasing the time devoted to practical skills did not improve the performance of ACLS students.

Identificación de antígenos inmunorreactivos de Leptospira interrogans / Identification of immunoreactive antigens of Leptospira interrogans

Se estudió un lote de 28 sueros de llama (Lama gama) de la provincia de Jujuy, Argentina, a fin de identificar antígenos inmunorreactivos contra Leptospira interrogans. Se utilizaron distintas preparaciones antigénicas de la bacteria para estudiar la inmunorreactividad mediante microaglutinación (MAT), ELISA y Western inmunoblot. Un pool de sueros bovinos positivos a la MAT fue empleado como control. Todos los sueros de llama fueron negativos mediante MAT e igual resultado se observó mediante ELISA. Dos de los 28 sueros de llama y el pool de sueros bovinos positivos, al ser evaluados por Western inmunoblot, arrojaron resultados positivos y permitieron identificar proteínas inmunorreactivas. Por MALDI-TOF se logró establecer que la proteína asociada a los dos sueros de llama inmunorreactivos era una flagelina periplásmica de Leptospira interrogans serovar Lai STR, mientras que la asociada al pool de sueros bovinos positivos a Leptospira sp. se trataba de una lipoproteína de la membrana externa de Leptospira interrogans serovar Ballum, LipL21. Estas proteínas podrían ser utilizadas en el diseño de un nuevo ELISA aplicado al diagnóstico temprano de leptospirosis, ya sea en distintos tipos de ganado como así también en reservorios silvestres.

A batch of 28 llama (Lama gama) sera from Jujuy province in Argentina was studied in order to identify immune reactive antigens to Leptospira interrogans. Different antigenic preparations from the bacterium were used to study the immune reactivity by the microagglutinattion (MAT), ELISA and Western immunoblot tests. A control pool of positive bovine sera was used. All the llama sera were negative to MAT as well as to ELISA. Two of the llama sera and the positive bovine sera pool rendered positive results when evaluated by Western immunoblot, allowing the identification of immune reactive proteins. These proteins were identified by MALDI-TOF. A periplasmic flagellin of Leptospira interrogans serovar Lai STR called FlaB1 was identified from the reactive llama sera, and an external membrane lipoprotein of Leptospira interrogans serovar Ballum called LipL21 was identified from the pool of bovine positive sera. These proteins could be used in a new ELISA applied to the early diagnosis of leptospirosis in different kind of cattle or wild reservoirs.

Eritema elevatum diutinum / Erythema elevatum diutinum

El eritema elevatum diutinum (EED) es una dermatosis crónica, poco frecuente, de causa desconocida, que al comienzo presenta histopatológicamente una vasculitis leucocitoclásticade la piel y luego evoluciona hacia una fibrosis. Presentamos una paciente con lesiones dermatológicas características y generalizadas de esta rara entidad, en la que se obtuvo una buena respuesta con la sulfonoterapia.

Erythema Elevatum Diutinum (EED) is a rare chronic cutaneous condition of unknow origin, that initially presents histological as leucocytoclastic vasculitis of the skin and later resolves with fibrosis. We present a patient of this uncommon condition with typical cutaneous lesions and widespread involvement. The disease was initially misinterpreted on clinical examination. Dapsone therapy was followed by a remarkable recovery.

Minor segmental wall motion abnormalities detected in patients with Chagas' disease have adverse prognostic implications

Recent data from our laboratory have shown that patients with the indeterminate form of Chagas' disease can have impairment of left ventricular contractility, as evaluated by the slope of the left ventricle end-systolic pressure-dimension relationship. We also showed that Chagas' disease patients with minimal baseline wall motion abnormalities detected by two-dimensional echocardiography have more intense contractility impairment when compared to patients with the indeterminate form of the disease without this abnormality. The prognostic implications of these findings have not been established. We evaluated 59 patients (37-76 years, mean = 55 years) with different clinical forms of Chagas' disease, who had normal left ventricular global systolic function at baseline (57.6 ± 6.9 percent) and who had at least one additional echo during clinical follow-up (0.4-17.6; mean 4.6 years). Group 1 consisted of 14 patients with minor baseline left ventricle wall motion abnormalities and group 2 consisted of 45 patients without these abnormalities. During follow-up, global left ventricle systolic function deterioration was observed in 10 group 1 patients (71.4 percent) and in only 10 group 2 patients (22.2 percent; P < 0.005). Age and duration of follow-up were not independent determinants of left ventricular function deterioration in these patients. The present data indicate that mild segmental left ventricular wall motion abnormalities are associated with worsening of systolic function in Chagas' disease patients who have normal baseline global systolic performance.

El genotipo de Mycobacterium bovis mayoritario en la Argentina lo es también en las islas Británicas: la tuberculosis bovina provino de Gran Bretaña? / [The genotype of the principal Mycobacterium bovis in Argentina is also that of the British Isles: did bovine tuberculosis come from Great Britain?]

Bovine tuberculosis is a highly prevalent animal disease in Argentina. In this work evidence was obtained showing that a major Mycobacterium bovis group in Argentina had been introduced with the bovine bulls imported from the United Kingdom at the end of the XIX century. This evidence came from two sources: historical, obtained by bibliographical references, and from laboratory results, using a molecular typing method called spoligotyping. These strains are also present in other countries that introduced cattle from the same origin.

Tratamiento laparoscópico de la coledocolitiasis: resultados / Laparoscopic treatment of the choledocholithiasis: results

Antecedentes: en la última década la colecistectomía laparoscópica se ha consolidado como el patrón oro del tratamiento de la litiasis biliar sintomática. Sin embargo, aún persiste la controversia de cuál es la mejor estrategia cuando existe coledocolitiasis asociada. Objetivo: evaluar los resultados del tratamiento laparoscópico en un tiempo en una serie consecutiva de pacientes operados por coledocolitiasis. Lugar de aplicación: Centro Público Terciario de Referencia. Diseño: estudio observacional prospectivo. Población: 178 pacientes con coledocolitiasis tratados entre enero de 1996 y enero de 2000. Método: se excluyeron los pacientes colecistectomatizados con litiasis residual y aquellos con colangitis aguda grave. Los 178 pacientes en el estudio fueron tratados inicialmente por vía laparoscópica en un tiempo. Siempre se trató de resolver inicialmente la coledocolitiasis por vía transcística; ante el fracaso se indicó la coledocotomía laparoscópica. Se indicó la conversión a cirugía abierta ante la imposibilidad de eliminar completamente los cálculos de la vía biliar o por complicaciones quirúrgicas o anestésicas. Resultados: fueron convertidos a cirugía abierta 17 pacientes (9,6 por ciento). De los 161 pacientes en los que se resolvió la coledocolitiasis en un tiempo por vía laparoscópica en 121 (75 por ciento) fue a través de la vía transcística y en los 40 restantes (25 por ciento) a través de una coledocotomía laparoscópica. En el grupo de pacientes en los que se utilizó la vía transcística hubo 2 complicaciones intraoperatorias leves por la utilización de las canastillas de Dormía y 3 complicaciones postoperatorias. Una bilirragia fugaz, una litiasis residual y un biloma por falla del clip del cístico. No hubo reoperaciones ni mortalidad. La utilización de la vía transcística fue del 75,2 por ciento y la efectividad del 91 por ciento. En los 40 pacientes en los que se realizó coledocotomía laparoscópica la operación se completó en 21 casos con cierre primario del colédoco (CPC), en 16 pacientes se colocó un tubo de Kehr y en 3 se realizó una coledocoduodenoanastomosis. Tres pacientes con CPC presentaron bilirragia postoperatoria que se agotó antes de las 72 hs y otro debió ser reoperado por laparotomía al 4§ día por coleperitoneo. Dos pacientes presentaron litiasis residual una después de CPC y otro después de coledocodrenaje con tubo de Kehr...

Mycobacterium bovis: polymerase chain reaction identification in bovine Lymphonode biopsies and genotyping in isolates from Southeast Brazil by spolygotyping and restriction fragment length polymorphism

Diagnosis of the Mycobacterium tuberculosis complex by direct PCR of mediastinal lymphnode DNA and microbiological tests were compared in cattle suspicious of bearing tuberculous-like lesions detected during slaughter. The PCR procedure applied on DNA samples (n=54) obtained by adding alpha -casein into the thiocyanate extraction mix was positive in 70 percent of the samples. PCR confirmed the identification of 23 samples (100 percent) that grew in culture, 9 samples (60 percent) that failed to grow in culture, plus 6 (37.5 percent) samples that resulted in growth of bacterial contaminants. Genotyping by IS6110-RFLP and DR-spoligotyping analysis of seven samples revealed the presence of several polimorphisms. Seven of the isolates contained multiple copies of IS6110, thus defining the existence of five singular genotypes

Expression of the Mycobacterium bovis P36 gene in Mycobacterium smegmatis and the baculovirus/insect cell system

In the present study we evaluated different systems for the expression of mycobacterial antigen P36 secreted by Mycobacterium bovis. P36 was detected by Western blot using a specific antiserum. The P36 gene was initially expressed in E. coli, under the control of the T7 promoter, but severe proteolysis prevented its purification. We then tried to express P36 in M. smegmatis and insect cells. For M. smegmatis, we used three different plasmid vectors differing in copy number and in the presence of a promoter for expression of heterologous proteins. P36 was detected in the cell extract and culture supernatant in both expression systems and was recognized by sera from M. bovis-infected cattle. To compare the expression level and compartmentalization, the MPB70 antigen was also expressed. The highest production was reached in insect cell supernatants. In conclusion, M. smegmatis and especially the baculovirus expression system are good choices for the production of proteins from pathogenic mycobacteria for the development of mycobacterial vaccines and diagnostic reagents

Biologia reprodutiva de éguas: estudo do ciclo estral e momento de ovulaçäo / Reproductive biology of the mare: oestrous cycle and ovulation time

No presente estudo, foram utilizadas 21 éguas, das quais em 11 Puros-Sangues Arabes (PSA) e 10 Cruza Arabes (CA), entre 3 e 11 anos de idade. Para identificaçäo do estro (cio) utilizaram-se os métodos de rufiaçäo e palpaçäo retal, sendo que as éguas foram rufiadas 3 vezes ao dia até o final do estro para determinaçäo de sua duraçäo. Independente do estágio do ciclo, todos os animais foram examinados pelo menos 3 vezes por semana. Nos diagnóstico do momento de ovulaçäo, as éguas foram examinadas às 8 h, 12 h e 16 h durante todo o período de estro, verificando-se as condiçöes ovarianas e foliculares. A duraçäo média do ciclo estral foi de 24,24ñ6,00 dias com 7,50ñ4,16 dias de estro. Observou-se que o início do estro foi mais freqüente às 12 h do que às 8 h ou 16 h e que as ovulaçöes ocorreram 75 por cento à noite, estando distribuídas de igual maneira nos dois ovários. Notou-se, também, que a fase estral terminou em 85 por cento dos casos 24 horas após a ovulaçäo

The rat thymus contains a heparin-binding factor that modulates steroidogenesis in the testis

Thymus development and function are under the influence of hormones secreted by the gonads and pituitary. On the other hand, thymurus is crucial for the development of reproductive capacitities in female and male rats and we have shown that a factor derived from the prepubertal rat thy mus has antigonadotropic effect in ovarian and testis cells in vitro. In the present paper we show that the rat thymic factor which modulates gonadotropin action in the gonads is an heparin-binding factor. This capacity was also used as a useful tool to obtain this activity from semipure extracts. An acetone extract was prepared from 15 day old male rats and subjected to molecular filtration chromatography. The activity, of those fractions was investigated in a testis cells biossay, by measuring testosterone secretion under basal and hCG-stimulation. Active fraction were processed in a heparin-Sepharose affinity column. We found that fractions that eluted with 0.6 and 2M NaCl/10mM Tris had biological specific activity. The electrophoretic procedure showed that the apparent molecular weight of the Heparin Sephadex binding factor is 60 kDa. Since this factor was obtained from a protein peak that eluted in the volume of carbonic anhidrase a dimerization process could be involved. Present results show that the rat thymus has an heparin-binding factor that interacts with hCG in testis cells. This factor could play an interesting role in the mutual influence between thymus and gonads.