Association between self-replicating calcifying nanoparticles and aortic stenosis: a possible link to valve calcification.

AIMS: Among various hypotheses proposed for pathological tissue calcification, recent evidence supports the possibility that self-replicating calcifying nanoparticles (CNPs) can contribute to such calcification. These CNPs have been detected and isolated from calcified human tissues, including blood vessels and kidney stones, and are referred to as nanobacteria. We evaluated calcific aortic valves for the presence of CNP. METHODS AND RESULTS: Calcific aortic valves were obtained from 75 patients undergoing surgical valve replacement. The control group was formed by eight aortic valves corresponding to patients with heart transplants. In the microbiology laboratory, valves were screened for CNP using a 4-6 weeks specific culture method. The culture for CNP was positive in 48 of the 75 valves with aortic stenosis (64.0%) in comparison with zero of eight (0%) for the control group (P = 0.0005). The observation of cultures by way of scanning electron microscopy highlighted the resemblance in size and morphology of CNP. CONCLUSION: Self-replicating calcific nanometer-scale particles, similar to those described as CNP from other calcific human tissues, can be cultured and visualized from calcific human aortic valves. This finding raises the question as to whether CNP contribute to the pathogenesis of the disease or whether they are only innocent bystanders.

[Seroprevalence of viral hepatitis in a representative general population of an urban public health area in Castilla y León (Spain)]. / Seroprevalencia de las hepatitis virales en población general representativa de una zona básica de salud urbana en Castilla y León.

INTRODUCTION: Viral hepatitis is a major social, health and economic problem worldwide, requiring strict epidemiological control. METHODS: This study presents the viral hepatitis seroprevalence in a representative sample from an urban health care area in Valladolid (Spain). RESULTS: Antibody prevalence was as follows: anti-HAV 52%; anti-HBc, 8.2%; anti-HCV, 1.1%; anti-HEV, 0.8%; and anti-HGV 5.8%. Prevalence of anti-HAV, anti-HBc and anti-HGV increased significantly with age (P < 0.005 in all cases). In individuals younger than 20, prevalence of anti-HAV was 3.8%, anti-HBc < 0.28% and anti-HGV 1.3%. In the 20-39 year-old group, seroprevalence against HAV was associated with low educational levels (P = 0.009) and with birth in other provinces (P = 0.016). Anti-HBc seroprevalence was mainly associated with three factors: prior hospitalization before 1990 (P = 0.002; OR 3.32 [1.48-7.42]); compulsory military service before 1990 (P < 0.0001; OR 37.33 [3.68-378.03]); and acupuncture treatments (P = 0.018; OR 57.75 [26.17-127.42]). Seroprevalence against HGV was associated with hospitalizations before 1990 (P = 0.019; OR = 2.969 [1.154-7.639]). Seropositive status to HCV revealed a transfusion history (2 cases), hospitalization (1 case) or drug addiction (1 case). Only one case among those seropositive to HEV had a history of a prior trip to a HEV-endemic area. CONCLUSIONS: Our study shows that the seroprevalences of viral hepatitis in a representative sample of urban population of Castille and Leon are similar to the seroprevalences observed in the rest of Spain and other developed countries, lower than the ones observed in the studies performed in Spain in the last 20 years due to the measures of prophylaxis that were taken.

Seroprevalencia de las hepatitis virales en población general representativa de una zona básica de salud urbana en Castilla y León / Seroprevalence of viral hepatitis in a representative general population of an urban public health area in Castilla y León (Spain)

Introducción. Las hepatitis virales constituyen uno de los principales problemas sociosanitarios y económicos a nivel mundial por lo que precisan un estrecho control epidemiológico. Métodos. En el presente trabajo estudiamos la seroprevalencia de las hepatitis virales una muestra representativa de la población de una zona básica de salud urbana en Valladolid (España). Resultados. La prevalencia de anticuerpos anti-VHA (AcVHA) fue del 52%, de HBcAc del 8,2%, de AcVHC del 1,1%, de AcVHE 0,8% y AcVHG 5,8%. La prevalencia de AcVHA, HBcAc y AcVHG aumenta significativamente con la edad (p < 0,005 en todos los casos). En menores de 20 años la prevalencia de AcVHA es del 3,8%, HBcAc < 0,28% y AcVHG 1,3%. En el grupo de edad de 20-39 años, la seroprevalencia frente al VHA se asocia con niveles educativos bajos (p 5 0,009) y con el nacimiento en otras provincias (p 5 0,016). La seroprevalencia de HBcAc se asocia principalmente con hospitalizaciones anteriores a 1990 (p 5 0,002; OR: 3,32 [1,48-7,42]), realización del servicio militar obligatorio anterior a 1990 (p < 0,0001; OR: 37,33 [3,68-378,03]) y prácticas de acupuntura (p 5 0,018; OR: 57,75 [26,17-127,42]). La seroprevalencia frente a VHG se asocia con hospitalizaciones antes de 1990 (p 5 0,019; OR: 2,969 [1,154-7,639]). Los seropositivos frente a VHC tenían antecedentes de transfusiones (2 casos) hospitalización (1 caso) o drogadicción (1 caso). De los seropositivos frente a VHE sólo un caso tenía antecedentes de viaje a zona endémica para VHE. Conclusiones. Nuestro estudio muestra que las seroprevalencias de las hepatitis virales en una muestra representativa de población urbana de Castilla y León son similares a las seroprevalencias obtenidas en el resto de España y de los países desarrollados, inferior a la observada en los estudios realizados en España en los últimos 20 años consecuencia de las medidas profilácticas adoptadas (AU)

Introduction. Viral hepatitis is a major social, health and economic problem worldwide, requiring strict epidemiological control. Methods. This study presents the viral hepatitis seroprevalence in a representative sample from an urban health care area in Valladolid (Spain). Results. Antibody prevalence was as follows: anti-HAV 52%; anti-HBc, 8.2%; anti-HCV, 1.1%; anti-HEV, 0.8%; and anti-HGV 5.8%. Prevalence of anti-HAV, anti-HBc and anti-HGV increased significantly with age (P < 0.005 in all cases). In individuals younger than 20, prevalence of anti-HAV was 3.8%, anti-HBc < 0.28% and anti-HGV 1.3%. In the 20-39 year-old group, seroprevalence against HAV was associated with low educational levels (P 5 0.009) and with birth in other provinces (P 5 0.016). Anti-HBc seroprevalence was mainly associated with three factors: prior hospitalization before 1990 (P 5 0.002; OR 3.32 [1.48-7.42]); compulsory military service before 1990 (P < 0.0001; OR 37.33 [3.68-378.03]); and acupuncture treatments (P 5 0.018; OR 57.75 [26.17-127.42]). Seroprevalence against HGV was associated with hospitalizations before 1990 (P 5 0.019; OR 5 2.969 [1.154-7.639]). Seropositive status to HCV revealed a transfusion history (2 cases), hospitalization (1 case) or drug addiction (1 case). Only one case among those seropositive to HEV had a history of a prior trip to a HEV-endemic area. Conclusions. Our study shows that the seroprevalences of viral hepatitis in a representative sample of urban population of Castille and Leon are similar to the seroprevalences observed in the rest of Spain and other developed countries, lower than the ones observed in the studies performed in Spain in the last 20 years due to the measures of prophylaxis that werw taken (AU)

Utility of an immunocapture-agglutination test and an enzyme-linked immunosorbent assay test against cytosolic proteins from Brucella melitensis B115 in the diagnosis and follow-up of human acute brucellosis.

The utility of an immunocapture-agglutination (Brucellacapt, Vircell SL, Granada, Spain) test and an enzyme-linked immunosorbent assay IgG, IgA, and IgM (ELISA-IgG, ELISA-IgA, ELISA-IgM) against cytosolic proteins from Brucella melitensis B115 (R) was compared with ELISA-IgG, ELISA-IgA, and ELISA-IgM against smooth lipopolysaccharide (S-LPS) from B. melitensis 16M (S), serum agglutination test (SAT), and Coombs test in the diagnosis and follow-up for 10 months of 51 patients with acute brucellosis. The sensitivities of ELISA tests against cytosolic proteins varied from 49.0 % for ELISA-IgG to 64.7% for ELISA-IgM and were lower than the sensitivities showed by ELISA S-LPS (from 88.2% to 92.2%), SAT (88.2%), Coombs (96.1%), and Brucellacapt (98.0%) tests. Specificity was over 93% in all cases. The evolutionary behavior of the SAT, Coombs, and Brucellacapt tests was similar. There was a decrease of between 20% and 40% in antibody titer in the 10th month of evolution after treatment. The evolutional curves of IgG, IgA, and IgM against cytosolic protein increased slightly till the eighth month. The specific IgM and IgA antibodies against protein fractions began to show a drop from the eighth month on, showing levels slightly lower than the initial sera values by the end of the 10th month. In this month, titers of specific IgG against proteins fractions remained higher than the titers showed by the initial sera.

Seroepidemiologic study of Leishmania infantum infection in Castilla-Leon, Spain.

Leishmaniasis has increased in importance in recent years because infection with human immunodeficiency virus (HIV) has emerged as a risk factor for this disease. However, the actual prevalence of leishmaniasis in the general population of Spain is unknown. We present a study of the seroprevalence of infection with Leishmania infantum in the general population of Castilla-Leon, Spain. A random sample of individuals presenting to health care clinics (4,825 sera) and of HIV-infected patients in the autonomous community of Castilla-Leon was collected in 1996. The sero-prevalence of antibodies to L. infantum was determined by an indirect enzyme immunoassay and found to be 4.9% in the general population. There was a significant increase in seroprevalence with age (P = 0.001), from 3.96% in those 14-20 years old to 7.2% in those > 70 years old. There were no significant differences between women and men (5.0% versus 4.9%; P = 0.9534). Seroprevalence was significantly higher in people from rural areas than in those from cities (6.0% versus 3.4%; P = 0.001). Patients infected with HIV had a seroprevalence for L. infantum of 64.0%. No differences were observed between women and men, and prevalence did not increase with age.

Susceptibility of the Acinetobacter calcoaceticus-A. baumannii complex to imipenem, meropenem, sulbactam and colistin.

The Acinetobacter calcoaceticus-Acinetobacter baumannii complex includes some of the most clinically relevant species of the genus Acinetobacter due to their capacity to cause epidemic nosocomial outbreaks as well as their increasing resistance to antibiotics. Susceptibility of Acinetobacter strains varies greatly depending on origin, thus highlighting the importance of local analyses of susceptibility profiles. Two hundred twenty-one strains of the A. calcoaceticus-A. baumannii complex were identified using biochemical tests and were biotyped. Strain susceptibility to imipenem, meropenem, colistin and sulbactam was studied using agar dilution. Eight different biotypes were found, type 1 accounting for 69.2% of the strains. MIC(50) and MIC(90) to imipenem, meropenem, colistin and sulbactam were 4 and 8 mg/l, 16 and 32 mg/l, 0.5 and 1mg/l, and 8 and 16 mg/l, with susceptibility rates of 64.3, 22.6, 98.2 and 73.8%, respectively. Biotype 1 was the most resistant. A statistically significant difference was observed for the mean MIC of the four predominant biotypes to imipenem, meropenem and sulbactam but not to colistin.

[Seroprevalence of infection due to Echinococcus granulosus in the population of Castilla and León (Spain)]. / Seroprevalencia de infección por Echinococcus granulosus en la población de Castilla y León.

INTRODUCTION: Human hydatidosis is a prevalent zoonotic disease in the Castilla y León region of Spain. The aim of this study is to investigate the seroprevalence of Echinococcus granulosus infection in this region. METHODS: We studied 4824 serum samples from a random, representative population of healthy individuals from each province of Castilla y León, obtained over one year. An indirect enzyme-immunoassay developed in our laboratory was used to determine the presence of IgG antibodies against Echinococcus granulosus in these samples. RESULTS: IgG antibodies against Echinococcus granulosus were detected in 3.4% (164/4824) of samples studied, with a range of 1.26% to 7.10%, depending on the province. Antibody seroprevalence increased significantly with age, but there was no significant sex-related difference (3.66% men vs. 3.14% women). CONCLUSION: The seroprevalence of Echinococcus granulosus infection in Castilla y León is still high. These data contribute to hydatidosis surveillance within the control program for this disease.

Seroprevalencia de infección por Echinococcus granulosus enla población de Castilla y León / Seroprevalence of infection due to Echinococcus granulosus in the population of Castilla and León (Spain)

INTRODUCCIÓN. La hidatidosis es una de las zoonosis más importantes en la Comunidad Autónoma de Castilla y León. Este estudio pretende conocer la seroprevalencia de infección por Echinococcus granulosus en dicha comunidad autónoma. MÉTODOS. Se han estudiado 4.824 muestras de suero pertenecientes a 4.824 personas seleccionadas de forma aleatoria y que constituían una muestra representativa de la población de las provincias de Castilla y León. En cada suero se estudió la presencia de anticuerpos de clase IgG frente a Echinococcus granulosus mediante una prueba de enzimoinmunoanálisis indirecto de fabricación propia. RESULTADOS. Se detectaron anticuerpos de clase IgG frente a Echinococcus granulosus en el 3,40 por ciento de los sueros estudiados (164 positivos de 4.824), oscilando entre el 1,26 y el 7,10 por ciento según la provincia de origen. La seroprevalencia de anticuerpos aumentaba significativamente con la edad. No se observaron diferencias estadísticamente significativas entre las seroprevalencias halladas en mujeres y en varones (3,14 por ciento frente a 3,66 por ciento). CONCLUSIÓN. La seroprevalencia de infección por E. granulosus en la Comunidad Autónoma de Castilla y León es todavía alta. Estos datos de seroprevalencia contribuyen a la vigilancia de la hidatidosis dentro de un programa control de esta enfermedad (AU)

Development of a dipstick dye immunoassay for diagnosing hydatidosis.

We have developed a kit to diagnose hydatidosis, based on the detection of specific antibodies. This disease, caused by larvae of the cestode Echinococcus granulosus, is a zoonosis commonly found throughout the world. The diagnostic kit discriminates between positive and negative cases in a way that is easily interpreted, i.e., positive cases form a colored band on an inert substrate. The results show test sensitivity to be some 94.87% and specificity 85.71%, which is comparable to those of the ELISA technique. The high sensitivity and specificity of this test affords the great advantage of speed in diagnosing this parasitosis. No other equipment or procedure is required.

Utility of a commercial immunoblot kit (BAG-Borrelia blot) in the diagnosis of the preliminary stages of Lyme disease.

The aim of this study was to evaluate the usefulness of a commercial immunoblot (IgG and IgM BAG-Borrelia blot) in the serologic diagnosis of the early stages of Lyme disease. A total of 42 sera from patients with Lyme disease (24 patients with localized early stage (LES) and 18 patients with disseminated early stage (DES)) and 129 sera from patients with non-Lyme diseases (specificity control sera) were studied. IgG anti-p41 from Borrelia burgdorferi s.l. was present in 95.2% of patients followed by anti-p41/I PBi (16.7%), anti-p100 (9.5%) and anti-OspA (9.5%). IgM anti-p41 was present in 66.7% of patients, p41/iPBi (54.8%) and OspC (33.3%). IgM against p100, OspA and OspC were more frequent in DES patients (16.7%, 27.8% and 44.4%) than in LES patients (0.0%, 4.2% and 25.0%). In 4.8% of the cases no IgG bands were present and in 26.2% no IgM bands were present. With the exception of isolated p41 bands (59.5%), no band pattern exceeded 17%. Using manufacturer's instructions, test sensitivity in diagnosis of the early stage of Lyme disease is 61.9%, specificity 98.4% and positive and negative predictive values 92.8% and 88.8% respectively. Applying the EUCALB 5, 6 or 7 rules sensitivity increased to 73.8% although specificity decreased to 89.9%. Of the 129 specific control sera, 41.8% presented IgG anti-p41 and 10.8% IgM anti-p41. Patients with non-Lyme diseases that presented more IgG and IgM bands were those patients with syphilis (88.2%), patients with anti-HIV antibodies (57.8%) and patients with anti-nuclear antibodies (ANA) (52.3%).

Serologic evidence of human infection by Francisella tularensis in the population of Castilla y León (Spain) prior to 1997.

Prior to an outbreak in Castilla y León in December 1997, tularaemia was practically non-existent in Spain. In this paper we studied the prevalence of antibodies against Francisella tularensis in a representative sample of the population (4825 people) from Castilla y León (Spain) in samples collected before this outbreak. Antibodies against F. tularensis were detected in nine (0.19%) of the 4825 sera, with antibody titres ranging from 1/20 to 1/160. Of these nine sera, one was positive in seroagglutination against Brucella. Seroagglutination against other bacteria (Yersinia enterocolitica O:9 and O:3 and Proteus OX19) was negative in all sera. Seroprevalence of antibodies in females was 0.20% and 0.17% in males; no statistically significant differences were found in prevalence in terms of sex, age or province.

[Prevalence of antibodies against Francisella tularensis in Castilla y León (Spain) before 1997]. / Prevalencia de anticuerpos frente a Francisella tularensis en la población de Castilla y León con anterioridad a 1997.

BACKGROUND AND OBJECTIVES: The aim of this paper was determine the prevalence of antibodies against Francisella tularensis in the representative sample of people from Castilla-León (Spain) before epidemic outbreak of end 1997. SUBJECTS AND METHOD: We obtain 4,825 sera (between april-1996 and april-1997) of people from Castilla-León. All sera were tested by a microagglutination technique to detect antibodies against Francisella tularensis. The positive sera were tested to determine cross-reaction with Brucella, Yersinia enterocolitica and Proteus in the tube agglutination tests. RESULTS: We detected antibodies against Francisella tularensis in 9 (0,19%) of the 4,825 sera. Only one serum from the 9 seropositive was positive in the tube agglutination against Brucella. None of the 9 sera were positive against the remaining bacterial antigen tested. CONCLUSIONS: In the people of Castilla-León before 1997 the prevalence of antibodies against Francisella tularensis was 0,19%.

Prevalencia de anticuerpos frente a Francisella tularensis en la población de Castilla y León con anterioridad a 1997 / Prevalence of antibodies against Francisella tularensis in Castilla y León (Spain) before 1997

FUNDAMENTO Y OBJETIVOS: Este trabajo pretende conocer la prevalencia de infección por Francisella tularensis en la población de Castilla y León previa al brote de tularemia humana de finales de 1997. SUJETOS Y MÉTODO: Se obtuvieron 4.825 sueros (entre abril de 1996 y abril de 1997) de residentes en Castilla y León. Se realizó una prueba de microaglutinación en placa para detectar anticuerpos anti-F. tularensis. En los sueros positivos se llevaron a cabo seroaglutinaciones en tubo frente a Brucella, Yersinia enterocolitica y Proteus. RESULTADOS: Se detectaron anticuerpos anti-F.tularensis en 9 (0,19 per cent) de los 4.825 sueros. De esos 9 sueros, uno fue positivo en la seroaglutinación frente a Brucella, siendo todos negativos frente a las otras bacterias. CONCLUSIONES: Antes de 1997 la seroprevalencia de anticuerpos anti-F. tularensis en la población de Castilla y León era baja (0,19 per cent) (AU)

[Genotypic resistance of human immunodeficiency virus in patients with virologic failure]. / Resistencias genotípicas del virus de la inmunodeficiencia humana en pacientes con fracaso virológico.

BACKGROUND: The aim of the present study is to asses the prevalence of resistances in a group of patients with virological failure establishing the relationship between the appearance of mutations and the given antiretroviral therapy along with other variables used in these patients follow-up. PATIENTS AND METHOD: Samples belonging to 88 patients were selected either with viral load levels above 30.000 copies/ml after reaching undetectable viral load levels, or with persistently detectable levels above 1.000 copies/ml. Resistances were tested by means of Line Probe Assay (LiPA). The history of patients' antiretroviral treatments was reviewed. RESULTS: Mutations were observed in 52,6% of cases for reverse transcriptase (RT) an in 81,8% for the protease genes, being T215Y and V82A the most frequently detected ones. Mutations coferring resistance to the given antiretrovirals appeared in 33 cases. No statistical significance was observed between the presence of mutations and the administered therapy. In the multivariate analysis we found for LiPA RT a greater risk of appearance of mutations according to patient motility (OR = 4,0). CONCLUSIONS: the prevalence of resistance mutations in patients with virologic failure is placed around 50% in both genes. A consensus in the definition of virologic failure in HIV infected patients is urged.

Resistencias genotípicas del virus de la inmunodeficiencia humana en pacientes con fracaso virológico / Genotypic resistance of human immunodeficiency virus in patients with virologic failure

FUNDAMENTO: El objetivo del presente estudio es establecer la prevalencia de resistencias en un grupo de pacientes en los que el tratamiento antirretroviral parece haber fracasado, analizando la relación entre la aparición de mutaciones y los regímenes terapéuticos seguidos, junto con otras variables asociadas al seguimiento. PACIENTES Y MÉTODO: Se seleccionaron muestras correspondientes a 88 pacientes con cargas virales superiores a 30.000 copias ARN/ml tras haber alcanzado valores indetectables, o cargas virales persistentemente detectables superiores a 1.000 copias ARN/ml. Se analizaron las resistencias mediante Line Probe Assay (LiPA) y se revisó la historia de tratamientos antirretrovirales de los pacientes. RESULTADOS: Se obtuvieron mutaciones en el 52,6 por ciento de los casos para el gen de la transcriptasa inversa (RT) y en un 51,8 por ciento para el de la proteasa, siendo la T215Y y la V82A las más frecuentes. En 33 casos hubo coincidencia entre la aparición de mutaciones que conferían resistencia para los fármacos que los pacientes recibían. No se encontró significación estadística entre la aparición de mutaciones y los tratamientos empleados en el seguimiento de los pacientes. En el análisis multivariante se encontró para LiPA RT, respecto a la movilidad, un riesgo 4,0 veces superior de aparición de mutaciones. CONCLUSIONES: La prevalencia de mutaciones de resistencia en pacientes en fracaso virológico se sitúa en torno al 50 por ciento de los casos en ambos genes. Es necesario establecer un consenso para definir el fracaso virológico en los pacientes con infección por el VIH. (AU)

Comparative study of two methods for RNA extraction prior to detection of resistance to human immunodeficiency virus type 1 with the line probe assay.

We evaluated two methods from Roche and Promega for RNA extraction prior to the genotypic detection of human immunodeficiency virus type 1 resistance by line probe assay (LiPA). Fifty plasma RNA extracts were processed in parallel by LiPA. Results obtained by the Roche method were superior in the proportion of amplified samples, the percentage of mutated samples, and band intensity.

Diagnóstico de laboratorio y evolución serológica de pacientes con tularemia / Laboratory diagnosis and serologic evolution of patients with tularemia

Fundamento: La tularemia era una enfermedad prácticamente inexistente en España hasta finales de 1997, cuando se declaró un brote epidémico en nuestra comunidad. El objetivo de nuestro trabajo ha sido estudiar los datos existentes sobre el diagnóstico microbiológico de 55 pacientes que sufrieron tularemia. Pacientes y métodos: Se obtuvieron 32 muestras para cultivo pertenecientes a 19 pacientes y 151 sueros correspondientes a 55 pacientes. El diagnóstico serológico se realizó mediante seroaglutinación en tubo y microaglutinación. En todos los sueros se realizó una seroaglutinación de Wright (SAW) y un test de Coombs frente a Brucella y seroaglutinaciones frente a Yersinia enterocolitica O:9, Yersinia enterocolitica O:3 y Proteus OX 19. Resultados: Se aisló F. tularensis en dos muestras (6,25 por ciento) de las 32 estudiadas. Se obtuvieron títulos mayores o iguales a 1/160 en el 78,2 por ciento y en el 74,5 por ciento de los sueros iniciales por sero-aglutinación en tubo y microaglutinación, respectivamente. La correlación entre las dos pruebas fue de 0,80 (p < 0,001). Se observó fenómeno de prozona en el 59,9 por ciento de los sueros, y reactividad cruzada frente a Brucella y Proteus OX19 en el 9,3 y el 22,8 por ciento, respectivamente. No se observó reactividad cruzada con Y. enterocolitica O:3 y O:9. Conclusiones: El cultivo de F. tularensis es poco sensible. La correlación obtenida entre la seroaglutinación en tubo y microaglutinación es buena. Ambas técnicas son útiles en el diagnóstico de la tularemia, con algunas ventajas de la microaglutinación sobre la aglutinación. (AU)