RESUMEN
Alpha-1 antitrypsin deficiency (AATD) arises due to inherited variants in SERPINA1, the AAT gene that impairs the production or secretion of this hepatocellular protein and leads to a gain-of-function liver proteotoxicity. Homozygous Pi*Z pathogenic variant (Pi*ZZ genotype) is the leading cause of severe AATD. It manifests in 2 to 10% of carriers as neonatal cholestasis and 20 to 35% of adults as significant liver fibrosis. Both children and adults may develop an end-stage liver disease requiring liver transplantation. Heterozygous Pi*Z pathogenic variant (Pi*MZ genotype) constitutes an established disease modifier. Our review summarizes the natural history and management of subjects with both pediatric and adult AATD-associated liver disease. Current findings from a phase 2 clinical trial indicate that RNA silencing may constitute a viable therapeutic approach for adult AATD. In conclusion, AATD is an increasingly appreciated pediatric and adult liver disorder that is becoming an attractive target for modern pharmacologic strategies.
Asunto(s)
Colestasis , Deficiencia de alfa 1-Antitripsina , Adulto , Humanos , Niño , Recién Nacido , Deficiencia de alfa 1-Antitripsina/complicaciones , Deficiencia de alfa 1-Antitripsina/genética , Deficiencia de alfa 1-Antitripsina/terapia , Cirrosis Hepática/genética , Cirrosis Hepática/complicaciones , Genotipo , Colestasis/complicacionesRESUMEN
Frozen berries have been repeatedly identified as vehicles for norovirus (NoV) transmission causing large gastroenteritis outbreaks. However, virus detection in berries is often hampered by the presence of RT-PCR-inhibiting substances. Here, several virus extraction methods for subsequent real-time RT-PCR-based NoV-RNA detection in strawberries were compared and optimized. NoV recovery rates (RRs) between 0.21 ± 0.13% and 10.29 ± 6.03% were found when five different artificially contaminated strawberry batches were analyzed by the ISO/TS15216-2 method indicating the presence of different amounts of RT-PCR inhibitors. A comparison of five different virus extraction methods using artificially contaminated strawberries containing high amounts of RT-PCR inhibitors revealed the best NoV RRs for the ISO/TS15216 method. Further improvement of NoV RRs from 2.83 ± 2.92% to 15.28 ± 9.73% was achieved by the additional use of Sephacryl(®)-based columns for RNA purification. Testing of 22 frozen strawberry samples from a batch involved in a gastroenteritis outbreak resulted in 5 vs. 13 NoV GI-positive and in 9 vs. 20 NoV GII-positive samples using the original ISO/TS15216 method vs. the extended protocol, respectively. It can be concluded that the inclusion of an additional RNA purification step can increase NoV detection by the ISO/TS15216-2 method in frozen berries containing high amounts of RT-PCR inhibitors.
Asunto(s)
Fragaria/química , Fragaria/virología , Alimentos Congelados/virología , Norovirus/aislamiento & purificación , ARN Viral/aislamiento & purificación , Reacción en Cadena en Tiempo Real de la Polimerasa , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Microbiología de Alimentos/métodos , Límite de Detección , Norovirus/genética , Inhibidores de la Síntesis del Ácido Nucleico/análisis , ARN Viral/genéticaRESUMEN
As an intermediate host of avian and human influenza A viruses (FLUAV) pigs may play a potential role in interspecies virus transmission and reassortment of viral genes including those conferring antiviral drug resistance. Porcine FLUAV isolated in Germany between 1989 and 2001 contains mutations in the M2 gene inducing amantadine resistance. No data exist on neuraminidase inhibitor (NAI) susceptibility of these porcine FLUAV. We studied the antiviral activity of NAI against seven selected H3N2 FLUAV isolated from pigs in Germany between 1982 and 1999. All isolates were susceptible towards oseltamivir and zanamivir in neuraminidase enzyme-inhibition assays. Both compounds inhibited virus spreading and reduced the virus yields and plaque size at low concentrations. Higher concentrations were necessary to reduce the plaque number. Two isolates that differed in glycosylation pattern of viral hemagglutinin (HA) showed markedly reduced drug susceptibility in cell culture-based assays.
Asunto(s)
Antivirales/farmacología , Subtipo H3N2 del Virus de la Influenza A/efectos de los fármacos , Neuraminidasa/antagonistas & inhibidores , Oseltamivir/farmacología , Zanamivir/farmacología , Amantadina/farmacología , Sustitución de Aminoácidos , Animales , Línea Celular , Farmacorresistencia Viral , Inhibidores Enzimáticos/farmacología , Alemania , Hemaglutinación por Virus , Glicoproteínas Hemaglutininas del Virus de la Influenza/química , Subtipo H3N2 del Virus de la Influenza A/aislamiento & purificación , Subtipo H3N2 del Virus de la Influenza A/metabolismo , Neuraminidasa/química , Neuraminidasa/metabolismo , PorcinosRESUMEN
A total of 26 rotavirus positive faecal samples of diarrhoeal foals, and 8 equine rotavirus isolates were examined. Viral RNA patterns were generated, G typing was performed by PCR, and a P[12]-specific DNA probe was developed for P typing. Furthermore, five equine rotavirus isolates were sequenced in the genomic regions coding for VP7 and part of VP4. Rotaviruses of genotype G3 P[12] were found in 22 faecal samples and G14 P[12] type could be found in 4 faecal samples. These findings confirm that in Germany G3 P[12] is the predominating type of equine rotaviruses.
Asunto(s)
Enfermedades de los Caballos/virología , Infecciones por Rotavirus/veterinaria , Rotavirus/clasificación , Animales , Secuencia de Bases , Southern Blotting/veterinaria , Diarrea/veterinaria , Diarrea/virología , Heces/virología , Genotipo , Alemania , Caballos , Datos de Secuencia Molecular , ARN Viral/química , ARN Viral/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinaria , Rotavirus/genética , Rotavirus/aislamiento & purificación , Infecciones por Rotavirus/virología , Análisis de Secuencia de ADN , Proteínas Virales/química , Proteínas Virales/genéticaRESUMEN
Noroviruses are important causes of gastroenteritis; however, due to a lack of sensitive detection methods, the distinct role of contaminated food in norovirus outbreaks remains unclear. Two published virus extraction procedures combined with real-time RT-PCR for the detection of norovirus from food inoculated experimentally were compared. The elution-precipitation method was most efficient in all food matrices tested showing detection limits of 20 RT-PCRU for lettuce and ham, and 200 RT-PCRU for raspberries. The average recovery rates were 23%, 7% and 24% for lettuce, raspberries and ham, respectively. The ultrafiltration method yielded detection limits of 200 RT-PCRU for lettuce and ham, and 2000 RT-PCRU for raspberries; recovery rates were 9%, 7%, 3%, respectively. Subsequently, food items implicated in a norovirus outbreak were examined by the elution-precipitation method. Virus recovery rates determined by using MS2 phage ranged from 1 to 20% depending on the food matrix. However, norovirus could not be detected in the food items examined. This negative result may be explained by a low virus titer and heterogeneous virus distribution, or by random selection of food samples that contained no norovirus. Both, proper sampling and virus extraction from foods may be improved further to identify vehicles of infection.
Asunto(s)
Infecciones por Caliciviridae/virología , Brotes de Enfermedades , Alimentos/virología , Gastroenteritis/virología , Norovirus/aislamiento & purificación , Virología/métodos , Preescolar , Humanos , Levivirus/aislamiento & purificación , ARN Viral/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Sensibilidad y Especificidad , Ultrafiltración/métodosRESUMEN
According to current scientific opinion the risk of human infection with H5N1 via preparation and consumption of poultry meat is negligible.This opinion has not yet been challenged by a formal risk assessment, due to the lack of empirical data. We have developed a scenario pathway model as a conceptual framework for a formal assessment of the H5N1 risk to humans through consumption of poultry meat and parameterise the model using information derived from expert opinions. The aim of this study was to investigate whether the notion of an overall negligible risk via the oral infection route is consistent with ad hoc data and expert opinions on the relevant parameters of the model. The model is mainly based on expert opinion. A stochastic Monte-Carlo simulation was conducted which took into consideration (amongst others) the exposure and infection of chicken (broiler and layer), turkeys, ducks and geese, the probabilities of detection prior to slaughter, virus survival and contamination during slaughter, as well as during the cutting and preparation of meat in commercial plants and in private households, respectively. The empirical consumption pattern for poultry meat in Germany was taken into account in the simulation. The results show that the risk for the individual consumer is practically zero whereas up to 23 cases per year in Germany might occur if the upper (more pessimistic) ranges of the expert opinions apply. The finding of a low but non-negligible risk to the population is discussed in relation to the epidemiological information available from recent outbreaks in South East Asia.
Asunto(s)
Contaminación de Alimentos , Subtipo H5N1 del Virus de la Influenza A , Gripe Aviar/transmisión , Gripe Humana/etiología , Carne/virología , Zoonosis , Animales , Seguridad de Productos para el Consumidor , Brotes de Enfermedades/prevención & control , Brotes de Enfermedades/veterinaria , Manipulación de Alimentos/métodos , Alemania , Humanos , Modelos Teóricos , Método de Montecarlo , Aves de Corral , Medición de Riesgo , Procesos EstocásticosRESUMEN
Connexins are known to play an essential role in the ischemic preconditioning (IP) of the heart; their functional role in this process, however, has not been clearly defined. For this reason, anesthetized rats were subjected to regional myocardial ischemia, with or without IP or reperfusion. In frozen sections of hearts, fluorescence immunohistochemical staining for connexin43 (Cx43) was performed. In contrast to undisturbed zones, tissue that had been subjected to ischemia revealed Cx43 immunostaining not only in the gap junctions but also in a conspicuous pattern in the free cellular membranes of the myocytes. In myocardium that was exposed to IP only, the ratio of immunofluorescence intensity in the free cellular membrane to that in the interior of the cell was 1.22 +/- 0.04 (ratio in non-ischemia-exposed area = 1.04 +/- 0.01). When 15 or 45 min of permanent ischemia followed IP, the effect became more evident (ratio = 1.31 +/- 0.03 and 1.46 +/- 0.03, respectively) and proved to be significantly greater than in the corresponding non-IP groups (ratio = 1.16 +/- 0.03 and 1.30 +/- 0.03, respectively, P < 0.01). Reperfusion led to an overall weakening of fluorescence intensities and a disappearance of the observed IP-specific differences. We conclude that IP initiates a redistribution of Cx43 from its natural position in the gap junctions toward the free plasma membrane, thereby improving the cell's chance of survival during the subsequent phase of prolonged ischemia by an unknown, supposedly gap junction-independent, mechanism.
Asunto(s)
Conexina 43/metabolismo , Precondicionamiento Isquémico Miocárdico , Isquemia Miocárdica/metabolismo , Miocardio/metabolismo , Enfermedad Aguda , Anestesia , Animales , Membrana Celular/metabolismo , Membrana Celular/fisiología , Electrofisiología , Técnica del Anticuerpo Fluorescente , Hemodinámica/fisiología , Masculino , Infarto del Miocardio/patología , Miocardio/patología , Ratas , Ratas WistarRESUMEN
This study was designed to gain insight into amantadine susceptibility of porcine influenza A viruses isolated in Germany between 1981 and 2001. The 12 studied H1N1, H1N2, and H3N2 porcine influenza virus strains were isolated in chicken eggs and passaged once in MDCK cells. Plaque reduction assays were applied to examine virus susceptibility to amantadine. Genotyping was used to confirm drug resistance. In the results of these antiviral studies, only 3 of the 12 isolates were shown to be amantadine-susceptible. All resistant strains contained the AA substitutions G16E, S31N, and R77Q in the membrane protein 2 (M2). Additionally, L27A was detected in two H1N1 strains. S31N and/or L27A are well-known amino acid substitutions in M2 that confer amantadine resistance. The role of the pig as an intermediate host of avian and human influenza A viruses, the possible involvement of genetic reassortment, and the high incidence of naturally amantadine-resistant porcine influenza A viruses suggest a real risk of emergence of amantadine resistant human viruses. Therefore, drug susceptibility monitoring appears to be warranted for effective application of those drugs.
Asunto(s)
Alphainfluenzavirus/efectos de los fármacos , Amantadina/farmacología , Antivirales/farmacología , Infecciones por Orthomyxoviridae/veterinaria , Enfermedades de los Porcinos/virología , Secuencia de Aminoácidos , Sustitución de Aminoácidos , Animales , Línea Celular , Embrión de Pollo , Farmacorresistencia Viral/genética , Genes Virales , Alemania , Subtipo H1N1 del Virus de la Influenza A/efectos de los fármacos , Subtipo H1N1 del Virus de la Influenza A/genética , Subtipo H3N2 del Virus de la Influenza A/efectos de los fármacos , Alphainfluenzavirus/genética , Alphainfluenzavirus/aislamiento & purificación , Datos de Secuencia Molecular , Infecciones por Orthomyxoviridae/virología , Filogenia , Alineación de Secuencia , Porcinos , Proteínas de la Matriz Viral/genética , Ensayo de Placa ViralRESUMEN
Traditionally, the classification of risk areas of tick-borne encephalitis (TBE) is based on the recording of autochthonous cases of the disease. In Germany, an extension of these areas over the years and an increasing virus prevalence in ticks have been observed in recent years. Registration of foci with autochthonous TBE cases, recording of disease incidence and virus prevalence in ticks are all proven epidemiological methods to characterize TBE risk areas. These data are necessary for a scientifically proven recommendation of TBE vaccines, and they need to be updated regularly. These epidemiological methods have advantages and disadvantages with respect to the risk assessment of TBE areas. Despite the fact that these methods are suitable for risk assessment in practice, disease incidence (new cases per year/100,000 inhabitants) and virus prevalence in questing ticks did not correlate. Using nested RT-PCR we were able to demonstrate that the prevalence of TBE virus (TBEV) in ticks removed from humans was significantly higher than in unfed, free-living Ixodes ricinus of the same area. The 561 ticks collected from humans in doctors' surgeries in Bavaria in 2002 were examined by nRT-PCR. The estimated overall virus prevalence in tested ticks was 8.8% (95% CI: 6.45-11.57%). The removed ticks examined were classified according to the sites of exposure of the patients in the individual districts. Peak values were measured in the district of Regen with 20.6% and in the district of Freyung-Grafenau with 18.3%. In recent studies on unfed I. ricinus (nymphs, adults), the average TBEV prevalence in ticks in Bavarian risk areas was between 0.5% and 2%.
Asunto(s)
Virus de la Encefalitis Transmitidos por Garrapatas/aislamiento & purificación , Encefalitis Transmitida por Garrapatas/epidemiología , Ixodes/virología , Animales , Vectores Arácnidos/virología , Encefalitis Transmitida por Garrapatas/virología , Alemania/epidemiología , Humanos , Incidencia , Ninfa/virología , Prevalencia , ARN Viral/análisis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores de RiesgoRESUMEN
We examined influenza virus strains of the subtype H3N2 from outbreaks of respiratory diseases in swine herds in Germany. Four different clusters can be distinguished when comparing parts of the HA1 gene from porcine H3N2 isolates analyzed between 1982 and 2001. Comparison between these clusters reveals a bp homology of less than 90%. In contrast, the homology within the clusters is between 93.7 and 100%. Each of these clusters was confined to a specific time period. For the NA gene an additional cluster is formed by the porcine H1N2 isolate. The findings that different subtypes and drift variants are circulating in the German pig population explain the emergence of new influenza virus variants and the need for continued surveillance of swine.
Asunto(s)
Subtipo H3N2 del Virus de la Influenza A , Virus de la Influenza A/genética , Virus de la Influenza A/aislamiento & purificación , Infecciones por Orthomyxoviridae/veterinaria , Infecciones del Sistema Respiratorio/veterinaria , Enfermedades de los Porcinos/virología , Animales , Genes Virales , Variación Genética , Alemania/epidemiología , Glicoproteínas Hemaglutininas del Virus de la Influenza/genética , Epidemiología Molecular , Neuraminidasa/genética , Infecciones por Orthomyxoviridae/epidemiología , Filogenia , ARN Viral/genética , ARN Viral/aislamiento & purificación , Infecciones del Sistema Respiratorio/epidemiología , Infecciones del Sistema Respiratorio/virología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADN , Homología de Secuencia de Ácido Nucleico , Porcinos , Enfermedades de los Porcinos/epidemiologíaRESUMEN
Viruses are increasingly important as etiological agents of gastrointestinal infections. Because of improved diagnostic methods, in particular, because of molecular biological techniques, viruses can be detected much more frequently as pathogens of foodborne diseases. Apart from the hepatitis A virus (HAV) the Norovirus (NLV) is becoming more significant. On an international level methods are developed to detect the amount of viruses which minimise PCR inhibitors and which are applicable in routine diagnostic. There are numerous routines available in the field of virus diagnostic in shellfish and oysters which are, however, not yet standardised and applicable in the whole of Europe. This review highlights viral agents their characteristics as well as food contaminated with viruses. While most of the detection methods are cited from the literature, own preliminary results of virus detection in mussels are discussed in addition.
Asunto(s)
Bivalvos/virología , Microbiología de Alimentos , Enfermedades Transmitidas por los Alimentos/diagnóstico , Virus de la Hepatitis A/aislamiento & purificación , Norovirus/aislamiento & purificación , Mariscos/virología , Animales , Infecciones por Caliciviridae/diagnóstico , Infecciones por Caliciviridae/virología , Enfermedades Transmitidas por los Alimentos/virología , Gastroenteritis/diagnóstico , Gastroenteritis/virología , Hepatitis A/diagnóstico , Hepatitis A/virología , Virus de la Hepatitis A/genética , Humanos , Norovirus/genética , ZoonosisRESUMEN
In Germany, 100-300 autochthonous clinical TBE cases have been recorded annually. There are high-risk areas in Bavaria and Baden-Wuerttemberg and ongoing low-risk areas in Hesse, Thuringia, and the Rhineland-Palatinate and single cases in Saxony. In order to be able to evaluate the epidemiological changes described here, it must be mentioned that a new definition of TBE risk areas was introduced on the district level in 1998 in Germany and in 2001 with the new Infection Protection Act (Infektionsschutzgesetz) which states that TBE is a notifiable disease. This led to the replacement of earlier surveillance systems and to many changes to data collection. In 1998 63 country and town districts were TBE risk areas, in 2001 79 and in 2002 86. There were new risk districts within Bavaria and Baden-Wuerttemberg and outside these regions in Thuringia, Hesse and the Rhineland-Palatinate. An interesting trend was observed in TBE epidemiology. The TBE incidence in Bavaria and Baden-Wuerttemberg has been stable on a high level for years; outside these areas it has steadily been climbing (Odenwald, Thuringia). On the basis of epidemiological data on TBE from the eastern part of Germany since 1960, it is obvious that major changes in virus activity in TBE risk areas also occurred in the past, the explanation of which has remained a matter for speculation. The epidemiological situation in the different risk areas for TBE in Germany was found to vary considerably, if one considers the surveillance data of the last 40 years. 1. Establishment of completely new low-risk areas. 2. Reactivation of formerly active areas with endemic latency. 3. High-risk areas with stable viral activity over long periods. 4. High-risk areas which have expanded and merged with low-risk areas. 5. High-risk areas which have developed into endemic areas or become inactive. High-risk TBE areas from 1960-1975 (i.e. Mecklenburg-Western Pomerania) have since completely disappeared. There were, at the same time, high-risk areas in Thuringia which had only become latent and have now obviously become active again. The Odenwald demonstrated growing virus activity in the 1990s. These changes in TBE activity in German risk areas over more than the last 40 years are presented schematically. This ongoing number of risk areas is certainly linked to the notification obligation and greater public awareness. Nevertheless, any effects of ecological and climatic changes on the natural foci cannot be ruled out nor can changes in human leisure behaviour. Local weather conditions also have a major effect on the TBE incidence. Warm and dry summers may cause low tick activities, rainy summers may lead to low exposure rates of human beings. Even changes in forms of agricultural production prompted by different political structures probably have an impact as do economic constraints which may lead to lower vaccination and higher exposure rates. Regular, systematic virus prevalence measurements from 1997 to 2002 in field-collected ticks in German high-risk areas do not indicate any risk increase nor do they suggest a downward trend. Studies on virus prevalence in questing versus partially engorged ticks indicate that we neither exactly know nor understand the real quantitative relations between the virus and the host. In a first study, virus prevalence in Ixodes ricinus removed from humans was examined. Humans which were exposed in some districts near Passau in Bavaria. In the autumn of 2001, virus prevalence of unengorged free-living nymphs (n = 820) in this area was 0.38 (0.08-1.1)% and of adults (n = 90) 1.17 (0.03-6.38)%. Surprisingly, virus prevalence in partially engorged ticks from the same area collected during the same period was significantly higher (nymphs, n = 86, 6.9% and adults, n = 129, 9.3%). Virus-positive partially engorged ticks were only found in districts known as risk areas. Nucleotide and deduced amino acid sequence data of the PCR products have confirmed the presence of virus prototype Neudoerfl only.
Asunto(s)
Virus de la Encefalitis Transmitidos por Garrapatas/crecimiento & desarrollo , Encefalitis Transmitida por Garrapatas/epidemiología , Ixodes/virología , Animales , Encefalitis Transmitida por Garrapatas/virología , Alemania/epidemiología , Humanos , Incidencia , Ixodes/crecimiento & desarrollo , Prevalencia , ARN Viral/química , ARN Viral/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , RiesgoRESUMEN
Foodborne infections with Campylobacter spp. are increasing, especially antibiotic resistant strains are emerging. Quinolone resistant isolates can cause failure of therapy in severe clinical infections. Molecular characterisation is needed for the detection of resistant variants of C. jejuni. Therefore 23 isolates from poultry and human medicine as well as three control strains were tested for their minimal inhibitory concentration, their Single-Strand-Conformation-Polymorphism (SSCP)-PCR pattern (a method for the detection of resistance determining point mutations), and their sequence of the quinolone resistance determining region (QRDR). Six different SSCP types could be identified: two types for quinolone resistant isolates and other types containing so called silent mutations without influence on the resistance. A genotypic monitoring of the quinolone resistance in C. jejuni can be useful for the early detection of new resistance variants. As a screening method for detection of point mutations in the QRDR the SSCP-PCR can be applied. Compared to other genotypic methods the SSCP-PCR is less time and cost consuming and needs only standard technical equipment.
Asunto(s)
Infecciones por Campylobacter/microbiología , Campylobacter jejuni/efectos de los fármacos , Microbiología de Alimentos , Mutación Puntual , Quinolonas/farmacología , Animales , Infecciones por Campylobacter/tratamiento farmacológico , Infecciones por Campylobacter/epidemiología , Campylobacter jejuni/genética , Pollos/microbiología , Farmacorresistencia Bacteriana , Humanos , Pruebas de Sensibilidad Microbiana , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Polimorfismo Conformacional Retorcido-Simple , Enfermedades de las Aves de Corral/microbiología , Análisis de Secuencia de ADNRESUMEN
Knowledge concerning the prevalence of the tick-borne encephalitis virus (TBEV) in wild living tick populations is very important for understanding the epidemiology of the disease and for immuno prophylactic strategy. In Germany high and low risk areas of TBE exist. In the years 1997-2000, 533 autochthonous clinical TBE cases were recorded, in the high-risk areas of Bavaria and Baden-Wuerttemberg 140 and 363, and in the low risk areas in Hesse (Odenwald) and Rhineland-Palatinate 22 and 8, respectively. Corresponding to these case reports we have measured the virus prevalence in free living ticks in these four risk areas and compared these findings with the situation in high-risk areas in Latvia. In the years 1997-2000, 2,797 clinical TBE cases were recorded in Latvia. For the studies in Germany, a total of 17,398 Ixodesricinus ticks (14,860 nymphs and 2,538 adults) were collected by flagging and examined for TBEV, in Latvia the corresponding numbers were 525 I. ricinus ticks (350 adults and 175 nymphs) and 281 I. persulcatus ticks (adults only). Information concerning annual and seasonal differences of the TBEV prevalence in natural TBE foci is not available in Germany. This paper is a continuation of the study (Süss et al., 1999), starting in 1997. We investigated every year, in May and September, the virus prevalence in ticks in high risk areas of Bavaria (8 foci) and Baden-Wuerttemberg (5 foci). A total of 15,400 ticks (13,100 nymphs and 2,300 adults) were examined for TBEV. The ticks were tested for the presence of TBEV-RNA using a sensitive, nested-RT-PCR. The virus prevalence in the Bavarian foci of the whole tick population ranged from 0.3 to 2.0% during these four years, in adults between 1.2 and 5.3% and in nymphs between 0.1 and 1.4%. In the high-risk areas of Baden-Wuerttemberg, in the Black Forest, the estimated virus prevalence rates of investigated ticks varied from 0.2 to 3.4%, in adults from 0 to 4.8%, and in nymphs from 0.2 to 3.4%. Using the same methods, we have also tested the low risk areas in the Odenwald (840 nymphs, 160 adults) and in Rhineland-Palatinate (920 nymphs, 78 adults). Ticks were collected in those areas where most TBE cases were registered. The virus prevalence in the Odenwald was 0% in adults and 0.5% in nymphs, whereas in ticks from Rhineland-Palatinate we have not found any positive PCR signal. Sequence data of the PCR products have shown that all strains in Germany were closely related to the central European virus prototype Neudoerfl. In I. ricinus ticks, collected in Riga county, the following virus prevalence rates were found: in females 2.4%, in males 3.7%, and in all adults 3.0%, in nymphs 2.4% and in the I. ricinus tick population examined 2.8%. The virus prevalence in I. persulcatus, collected in the eastern parts of Latvia was 6% in females, 4% in males and 5% in all adults. All the PCR products were sequenced and a phylogenetic tree was constructed. Studies in natural foci of TBE in Latvia have shown that I. ricinus carried the central European virus subtype (prototype Neudoerfl) whereas in I. persulcatus two strains have been found, the central European virus subtype (prototype Neudoerfl) and the Siberian virus subtype (prototype Vasilchenko). Sequences of the Far Eastern subtype have not been detected yet.
Asunto(s)
Vectores Arácnidos/virología , Virus de la Encefalitis Transmitidos por Garrapatas/aislamiento & purificación , Encefalitis Transmitida por Garrapatas/epidemiología , Ixodes/virología , Animales , Virus de la Encefalitis Transmitidos por Garrapatas/clasificación , Virus de la Encefalitis Transmitidos por Garrapatas/genética , Encefalitis Transmitida por Garrapatas/diagnóstico , Alemania/epidemiología , Humanos , Letonia/epidemiología , Estudios Longitudinales , Ninfa/virología , Prevalencia , ARN Viral/análisis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Factores de Riesgo , Estaciones del AñoRESUMEN
To investigate the localization of the earliest damage in ischemic and ischemic-reperfused myocardium, anesthetized rats were subjected to coronary occlusion for 15, 30, 45, or 90 min. One-half of the animals in each group had no reperfusion, whereas the other half was reperfused for 14 min. With the use of histological methods, preferentially in the periphery of the area at risk, localized zones were detected that lacked the hypoxia-specific increase in NADH fluorescence. The extent of these areas displaying injured tissue was found to be significantly smaller in the ischemic-nonreperfused hearts than in the ischemic-reperfused organs (15-min ischemia: 0.22 +/- 0.12% vs. 43.0 +/- 5.0%; 30-min ischemia: 5.7 +/- 2.7% vs. 64.6 +/- 2.9%; 45-min ischemia: 5.6 +/- 1.2% vs. 66.0 +/- 7.5%; 90-min ischemia: 39.3 +/- 5.5% vs. 86.7 +/- 1.8% of the area at risk). The results point to a localized initiation of the damage close to the surrounding oxygen-supplied tissue during ischemia and an expansion of this injury by intercellular actions into yet-intact areas upon reperfusion.