RESUMEN
Polymerase chain reaction (PCR) detection has become the gold standard for diagnosis and typing of enterovirus (EV) and human parechovirus (HPeV) infections. Its effectiveness depends critically on using the appropriate sample types and high assay sensitivity as viral loads in cerebrospinal fluid samples from meningitis and sepsis clinical presentation can be extremely low. This study evaluated the sensitivity and specificity of currently used commercial and in-house diagnostic and typing assays. Accurately quantified RNA transcript controls were distributed to 27 diagnostic and 12 reference laboratories in 17 European countries for blinded testing. Transcripts represented the four human EV species (EV-A71, echovirus 30, coxsackie A virus 21, and EV-D68), HPeV3, and specificity controls. Reported results from 48 in-house and 15 commercial assays showed 98% detection frequencies of high copy (1000 RNA copies/5 µL) transcripts. In-house assays showed significantly greater detection frequencies of the low copy (10 copies/5 µL) EV and HPeV transcripts (81% and 86%, respectively) compared with commercial assays (56%, 50%; P = 7 × 10-5 ). EV-specific PCRs showed low cross-reactivity with human rhinovirus C (3 of 42 tests) and infrequent positivity in the negative control (2 of 63 tests). Most or all high copy EV and HPeV controls were successfully typed (88%, 100%) by reference laboratories, but showed reduced effectiveness for low copy controls (41%, 67%). Stabilized RNA transcripts provide an effective, logistically simple and inexpensive reagent for evaluation of diagnostic assay performance. The study provides reassurance of the performance of the many in-house assay formats used across Europe. However, it identified often substantially reduced sensitivities of commercial assays often used as point-of-care tests.
Asunto(s)
Infecciones por Enterovirus/diagnóstico , Enterovirus/clasificación , Parechovirus/clasificación , Infecciones por Picornaviridae/diagnóstico , ARN Viral/genética , Infecciones por Enterovirus/virología , Europa (Continente) , Dosificación de Gen , Humanos , Meningitis Viral/diagnóstico , Tipificación Molecular , Infecciones por Picornaviridae/virología , Juego de Reactivos para Diagnóstico , Reacción en Cadena en Tiempo Real de la Polimerasa , Reproducibilidad de los Resultados , Sensibilidad y EspecificidadRESUMEN
Paediatric asthma exacerbations are often caused by rhinovirus (RV). Moreover, 25(OH)-vitamin D3 (VitD3) deficiency during infancy was found associated with asthma. Here, we investigated the innate immune responses to RV and their possible modulation by 25(OH)-VitD3 serum levels in a preschool cohort of children with and without asthma. The innate lymphoid cell type 2 (ILC2)-associated marker, ST2, was found up-regulated in the blood cells of asthmatic children with low serum levels of 25(OH)-VitD3 in the absence of RV in their airways. Furthermore, in blood cells from control and asthmatic children with RV in their airways, soluble (s) ST2 (sST2) protein was found reduced. Asthmatic children with low 25(OH)-VitD3 in serum and with RV in vivo in their airways at the time of the analysis had the lowest sST2 protein levels in the peripheral blood compared to control children without RV and high levels of 25(OH)-VitD3. Amphiregulin (AREG), another ILC2-associated marker, was found induced in the control children with RV in their airways and low serum levels of 25(OH)-VitD3. In conclusion, the anti-inflammatory soluble form of ST2, also known as sST2, in serum correlated directly with interleukin (IL)-33 in the airways of asthmatic children. Furthermore, RV colonization in the airways and low serum levels of 25(OH)-VitD3 were found to be associated with down-regulation of sST2 in serum in paediatric asthma. These data indicate a counter-regulatory role of 25(OH)-VitD3 on RV-induced down-regulation of serum sST2 in paediatric asthma, which is relevant for the therapy of this disease.
Asunto(s)
Asma/inmunología , Colecalciferol/sangre , Resfriado Común/inmunología , Proteína 1 Similar al Receptor de Interleucina-1/metabolismo , Leucocitos Mononucleares/fisiología , Sistema Respiratorio/metabolismo , Rhinovirus/inmunología , Células Cultivadas , Niño , Preescolar , Estudios de Cohortes , Progresión de la Enfermedad , Femenino , Humanos , Inmunidad Innata , Proteína 1 Similar al Receptor de Interleucina-1/genética , Interleucina-33/metabolismo , Leucocitos Mononucleares/virología , Masculino , Regulación hacia ArribaRESUMEN
Children with rhinovirus-induced severe early wheezing have an increased risk of developing asthma later in life. The exact molecular mechanisms for this association are still mostly unknown. To identify potential changes in the transcriptional and epigenetic regulation in rhinovirus-associated atopic or nonatopic asthma, we analyzed a cohort of 5-year-old children (n = 45) according to the virus etiology of the first severe wheezing episode at the mean age of 13 months and to 5-year asthma outcome. The development of atopic asthma in children with early rhinovirus-induced wheezing was associated with DNA methylation changes at several genomic sites in chromosomal regions previously linked to asthma. The strongest changes in atopic asthma were detected in the promoter region of SMAD3 gene at chr 15q22.33 and introns of DDO/METTL24 genes at 6q21. These changes were validated to be present also at the average age of 8 years.
Asunto(s)
Asma/etiología , Asma/genética , D-Aspartato Oxidasa/genética , Infecciones por Picornaviridae/complicaciones , Ruidos Respiratorios/etiología , Rhinovirus , Proteína smad3/genética , Niño , Preescolar , Metilación de ADN , Epigénesis Genética , Femenino , Finlandia , Estudios de Seguimiento , Hospitales Universitarios , Humanos , Lactante , Masculino , Metiltransferasas/metabolismo , TranscriptomaRESUMEN
Some studies have assessed the efficacy of influenza vaccination in children separately for moderate-to-severe and any influenza, but the definition used for identifying children with moderate-to-severe illness has not been validated. We analyzed clinical and socioeconomic data from two prospective cohort studies of respiratory infections among children aged ≤13 years (four influenza seasons, 3,416 child-seasons of follow-up). We categorized children with laboratory-confirmed influenza into two mutually exclusive groups of moderate-to-severe and mild influenza using the previously proposed criteria. We obtained the data for the analyses from structured medical records filled out by the study physicians and from daily symptom cards filled out by the parents. Of 434 cases of influenza, 217 (50 %) were classified as moderate-to-severe and 217 (50 %) as mild. The mean duration of fever was 4.0 days in children with moderate-to-severe influenza and 3.1 days in those with milder illness (P < 0.0001). Antibiotics were prescribed to 111 (51 %) children with moderate-to-severe and to ten (5 %) children with mild influenza (P < 0.0001). The rates of parental work absenteeism were 184 days per 100 children with moderate-to-severe influenza and 135 days per 100 children with mild influenza (P = 0.02). The corresponding rates of children's own absenteeism from day care or school were 297 and 233 days respectively per 100 children (P = 0.006). Categorization of children into groups with moderate-to-severe and mild influenza is meaningful, and it identifies children in whom the clinical and socioeconomic impact of influenza is highest. Illness severity should be considered when assessing influenza vaccine effectiveness in children.
Asunto(s)
Gripe Humana/diagnóstico , Gripe Humana/epidemiología , Absentismo , Adolescente , Niño , Preescolar , Servicios Médicos de Urgencia , Femenino , Hospitalización , Humanos , Lactante , Virus de la Influenza A , Gripe Humana/virología , Betainfluenzavirus , Masculino , Fenotipo , Instituciones Académicas , Índice de Severidad de la Enfermedad , Factores SocioeconómicosRESUMEN
BACKGROUND: The role of viral infections in the etiology of severe community-acquired pneumonia (SCAP) was prospectively evaluated from 2008 to 2012 at a university-level intensive care unit. METHODS: Clinical data and microbiological tests were assessed: blood cultures, urine pneumococcal and legionella antigens, Mycoplasma pneumoniae and Chlamydia pneumoniae antibodies from paired serums, and respiratory virus detection by multiplex, real-time polymerase chain reaction (PCR) from nasopharyngeal swabs and lower tracheal specimens via intubation tube. RESULTS: Of 49 mechanically ventilated SCAP patients (21 men and 28 women; median age, 54 years), the etiology was identified in 45 cases (92%). There were 21 pure bacterial infections (43%), 5 probably pure viral infections (10%), and 19 mixed bacterial-viral infections (39%), resulting in viral etiology in 24 patients (49%). Of 26 viruses, 21 (81%) were detected from bronchial specimens and 5 (19%) from nasopharyngeal swabs. Rhinovirus (15 cases, 58%) and adenovirus (4 cases, 15%) were the most common viral findings. The bacterial-viral etiology group had the highest peak C-reactive protein levels (median, 356 [25th-75th percentiles, 294-416], P = .05), whereas patients with probably viral etiology had the lowest peak procalcitonin levels (1.7 [25th-75th percentiles, 1.6-1.7]). The clinical characteristics of pure bacterial and mixed bacterial-viral etiologies were comparable. Hospital stay was longest among the bacterial group (17 vs 14 days; P = .02). CONCLUSIONS: Viral findings were demonstrated in almost half of the SCAP patients. Clinical characteristics were similar between the pure bacterial and mixed bacterial-viral infections groups. The frequency of viral detection depends on the availability of PCR techniques and lower respiratory specimens.
Asunto(s)
Infecciones Comunitarias Adquiridas/epidemiología , Infecciones Comunitarias Adquiridas/virología , Neumonía Viral/epidemiología , Neumonía Viral/virología , Virus/clasificación , Virus/aislamiento & purificación , Adulto , Anciano , Anciano de 80 o más Años , Estudios de Cohortes , Infecciones Comunitarias Adquiridas/patología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa Multiplex , Neumonía Viral/patología , Estudios Prospectivos , Reacción en Cadena en Tiempo Real de la Polimerasa , Respiración Artificial , Adulto JovenRESUMEN
BACKGROUND: The relationships between tonsillar immune responses, and viral infection and allergy are incompletely known. OBJECTIVE: To study intratonsillar/nasopharyngeal virus detections and in vivo expressions of T-cell- and innate immune response-specific cytokines, transcription factors, and type I/II/III interferons in human tonsils. METHODS: Palatine tonsil samples were obtained from 143 elective tonsillectomy patients. Adenovirus, bocavirus-1, coronavirus, enteroviruses, influenza virus, metapneumovirus, parainfluenza virus, rhinovirus, and respiratory syncytial virus were detected using PCR. The mRNA expression levels of IFN-α, IFN-ß, IFN-γ, IL-10, IL-13, IL-17, IL-28, IL-29, IL-37, TGF-ß, FOXP3, GATA3, RORC2, and Tbet were directly analyzed by quantitative RT-PCR. RESULTS: Fifty percentage of subjects reported allergy, 59% had ≥1 nasopharyngeal viruses, and 24% had ≥1 intratonsillar viruses. Tonsillar virus detection showed a strong negative association with age; especially rhinovirus or parainfluenza virus detection showed positive association with IFN-γ and Tbet expressions. IL-37 expression was positively associated with atopic dermatitis, whereas IFN-α, IL-13, IL-28, and Tbet expressions were negatively associated with allergic diseases. Network analyses demonstrated strongly polarized clusters of immune regulatory (IL-10, IL-17, TGF-ß, FOXP3, GATA3, RORC2, Tbet) and antiviral (IFN-α, IFN-ß, IL-28, IL-29) genes. These two clusters became more distinctive in the presence of viral infection or allergy. A negative correlation between antiviral cytokines and IL-10, IL-17, IL-37, FOXP3, and RORC2 was observed only in the presence of viruses, and interestingly, IL-13 strongly correlated with antiviral cytokines. CONCLUSIONS: Tonsillar cytokine expression is closely related to existing viral infections, age, and allergic illnesses and shows distinct clusters between antiviral and immune regulatory genes.
Asunto(s)
Tonsila Palatina/inmunología , Tonsila Palatina/virología , Virosis/inmunología , Adolescente , Adulto , Niño , Análisis por Conglomerados , Citocinas/genética , Femenino , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Redes Reguladoras de Genes , Humanos , Masculino , Tonsila Palatina/metabolismo , Factores de Transcripción/genética , Transcriptoma , Virosis/genética , Adulto JovenRESUMEN
The number of cases of Mycoplasma pneumoniae infection detected by laboratory-based surveillance increased in Finland in late 2010. During 2011, the number of cases was four times higher than during the previous epidemic in 2005. The 2011 epidemic affected mostly school-age children. The increased number of cases was probably not due to changes in laboratory procedures, but public interest may have had an effect, since the number of Google queries followed closely the epidemic curve.
Asunto(s)
Epidemias/estadística & datos numéricos , Mycoplasma pneumoniae/aislamiento & purificación , Neumonía por Mycoplasma/epidemiología , Vigilancia de la Población/métodos , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Preescolar , Femenino , Finlandia/epidemiología , Humanos , Incidencia , Lactante , Recién Nacido , Masculino , Persona de Mediana Edad , Neumonía por Mycoplasma/diagnóstico , Adulto JovenRESUMEN
The purpose of this study was to examine the association between bacterial colonization/infection and respiratory outcomes in children younger than 3 years old who were hospitalized for their first wheezing episode. This was an observational study. The primary outcome was hospitalization time and the secondary outcomes included relapses within 2 months and time to recurrent wheezing (i.e. three physician confirmed wheezing episodes) within 12 months. Bacterial antibody assays for Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis, Mycoplasma pneumoniae and Chlamydia pneumoniae were studied as well as nasopharyngeal bacterial culture for the three former and urine pneumococcal antigen. Nasopharyngeal bacterial culture was positive in 31/52 (60%) children, serologic evidence of bacterial infection was found in 17/96 (18%) children, urine pneumococcal antigen was positive in 24/101 (24%), and any bacterial detection method was positive in 53/106 (50%) children. The children with positive nasopharyngeal bacterial culture had longer duration of hospitalization (hazard ratio 2.4) and more often relapsed within two months than those with negative culture (odds ratio 7.3). In this study, half of the first time wheezing children had bacterial colonization or symptomatic or asymptomatic bacterial infection. The bacterial colonization (i.e. positive nasopharyngeal bacterial culture) was associated with longer duration of hospitalization and higher risk of recurrent wheezing.
Asunto(s)
Infecciones Bacterianas/microbiología , Portador Sano/microbiología , Nasofaringe/microbiología , Ruidos Respiratorios , Anticuerpos Antibacterianos/sangre , Bacterias/aislamiento & purificación , Infecciones Bacterianas/epidemiología , Portador Sano/epidemiología , Preescolar , Femenino , Hospitalización , Humanos , Lactante , Tiempo de Internación , Masculino , Recurrencia , Medición de RiesgoRESUMEN
OBJECTIVE: Cytokine release syndrome is suggested to be the most important mechanism triggering acute respiratory distress syndrome and end organ damage in COVID-19. The severity of disease may be measured by different biomarkers. METHODS: We studied markers of inflammation and coagulation as recorded in 29 patients on admission to the hospital in order to identify markers of severe COVID-19 and need of ICU. RESULTS: Patients who were eventually admitted to ICU displayed significantly higher serum levels of interleukin-6 (IL-6), C-reactive protein (CRP), and procalcitonin. No statistical differences were found between the groups in median levels of lymphocytes, D-dimer or ferritin. CONCLUSIONS: IL-6 and CRP were the strongest predictors of severity in hospitalized patients with COVID-19.
Asunto(s)
COVID-19/sangre , COVID-19/diagnóstico , Interleucina-6/sangre , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Biomarcadores/sangre , Femenino , Humanos , Masculino , Persona de Mediana Edad , Índice de Severidad de la Enfermedad , Adulto JovenRESUMEN
BACKGROUND: Data on the link between atopy and viral wheeze are limited. AIM: To evaluate the association between IgE sensitization and viral infection in wheezing children. METHODS: This is an observational study in hospitalized wheezing children (n = 247; median age 1.6 ; interquartile range 1.1, 2.9). Eighteen respiratory viral infections were studied using all available methods. A specific immunoglobulin E (IgE) sensitization for common food and aeroallergens and other atopy-related variables including total IgE, blood and nasal eosinophils, exhaled nitric oxide, eczema and atopic eczema, parental allergy and asthma, number of wheezing episodes, positive asthma predictive index or asthma and use of inhaled corticosteroid were correlated with specific viral etiology. RESULTS: Atopy was closely associated with sole rhinovirus etiology (n = 58) but not with sole respiratory syncytial virus, sole enterovirus, sole human bocavirus, sole other virus, mixed viral, or virus negative etiology. The number of sensitizations was particularly associated with sole rhinovirus etiology (odds ratio 4.59; 95% confidence interval 1.78, 11.8; adjusted to age and sex), followed by aeroallergen sensitization (respectively; 4.18; 2.00, 8.72), total IgE level (2.06; 1.32, 3.21), food allergen sensitization (2.02; 1.08, 3.78), and nasal eosinophil count (1.52; 1.08, 2.13). CONCLUSIONS: According to our data, allergic sensitization is positively linked to rhinovirus-, but not other virus-, associated wheezing and calls attention for studies to test rhinovirus-associated wheezing as a part of asthma risk indices.
Asunto(s)
Hipersensibilidad/epidemiología , Infecciones por Picornaviridae/epidemiología , Rhinovirus/inmunología , Alérgenos/inmunología , Antígenos Virales/inmunología , Recuento de Células , Preescolar , Femenino , Humanos , Hipersensibilidad/sangre , Hipersensibilidad/fisiopatología , Hipersensibilidad/virología , Inmunización , Inmunoglobulina E/sangre , Lactante , Masculino , Infecciones por Picornaviridae/sangre , Infecciones por Picornaviridae/fisiopatología , Infecciones por Picornaviridae/virología , Ruidos Respiratorios , Rhinovirus/patogenicidad , Factores de RiesgoRESUMEN
BACKGROUND: Acute heart failure is a potentially fatal manifestation of viral myocarditis. Development of myocardial damage in myocarditis involves cardiomyocyte apoptosis. Levosimendan is a novel calcium sensitizing inotropic agent with anti-apoptotic properties. We studied the feasibility of inotropic treatment with levosimendan and its effects on apoptosis in experimental acute heart failure caused by coxsackievirus myocarditis. MATERIALS AND METHODS: Adolescent BALB/c mice were infected with myocarditic Woodruff variant of coxsackievirus B3 (2 x 10(4) plaque-forming units). Mice were randomized into those receiving levosimendan 0.33 mg kg(-1) (total dose 1 mg kg(-1) day(-1)) (n = 20) or vehicle (n = 19) given orally by gauge three times a day for 7 days after infection. Left ventricular function was evaluated by transthoracic echocardiography and the mice were euthanized on day 7. Histopathology, amount of virus in the heart (virus titration assay) and cardiomyocyte apoptosis (TUNEL assay) were studied. Uninfected untreated control mice were also studied. RESULTS: Infection resulted in histopathologically severe myocarditis and significant impairment of left ventricular function. Levosimendan treatment significantly improved ventricular function (fractional shortening 0.32 +/- 0.04 vs. 0.23 +/- 0.05, P = 0.005; contractility 0.60 +/- 0.12 vs. 0.39 +/- 0.14, P = 0.007 and myocardial performance index 0.36 +/- 0.06 vs. 0.62 +/- 0.15, P < 0.0001) compared with vehicle. Levosimendan also reduced cardiomyocyte apoptosis (0.26 +/- 0.08% vs. 0.44 +/- 0.15% in vehicle, P = 0.008), but did not have an effect on areas of myocardial necrosis or inflammation, or the amount of virus in the heart. Levosimendan treatment did not affect mortality (total mortality 63%). CONCLUSIONS; Levosimendan improves ventricular function and inhibits cardiomyocyte apoptosis; therefore, it is suggested as a potentially feasible therapy in acute heart failure caused by viral myocarditis.
Asunto(s)
Cardiotónicos/farmacología , Infecciones por Coxsackievirus/patología , Insuficiencia Cardíaca/patología , Hidrazonas/farmacología , Miocarditis/patología , Miocardio/patología , Piridazinas/farmacología , Animales , Infecciones por Coxsackievirus/tratamiento farmacológico , Insuficiencia Cardíaca/tratamiento farmacológico , Masculino , Ratones , Ratones Endogámicos BALB C , Miocarditis/tratamiento farmacológico , Miocarditis/virología , Miocitos Cardíacos/patología , Simendán , Función Ventricular Izquierda/efectos de los fármacosRESUMEN
BACKGROUND: Autoantibodies against various endogenous proteins are found in myocarditis. Troponin autoantibodies are detected in patients with chronic dilated cardiomyopathy, but their presence in myocarditis remains unknown. We set out to study the presence of troponin autoantibodies in experimental viral myocarditis. MATERIALS AND METHODS: BALB/c mice infected with coxsackievirus B3 Nancy strain were followed-up at days 1-7 and 2, 4, 8 and 12 weeks after infection. Levels of circulating cardiac troponin I and circulating troponin autoantibodies were measured. Transthoracic echocardiography was performed. Myocarditis was histopathologically graded and cardiomyocyte apoptosis was quantified (TUNEL). RESULTS: Histopathologically relatively mild acute myocarditis followed by persistent cardiomyocyte damage was observed. Rate of cardiomyocyte apoptosis was the highest on day 5 (0.16 +/- 0.01% vs. 0.03 +/- 0.01% in controls, P < 0.001). Circulating troponin I levels were increased to day 5 (45.2 +/- 6.5 ng mL(-1), P < 0.005 vs. controls). Troponin autoantibodies were detected from 2 weeks after infection (20% of animals had autoantibodies at 2 weeks, 60% at 4 and 8 weeks and 20% at 12 weeks, P < 0.05 vs. controls). Fractional shortening remained decreased after acute myocarditis (0.36 +/- 0.02 at 4 weeks, 0.30 +/- 0.02 at 8 and 12 weeks vs. 0.41 +/- 0.01 before infection, P < 0.01) parallel to development of troponin autoantibodies. CONCLUSION: Troponin autoantibodies are formed in experimental virus induced myocarditis following troponin I release and cardiomyocyte apoptosis. The definite role of these autoantibodies remains to be further characterized.
Asunto(s)
Apoptosis/inmunología , Autoanticuerpos/inmunología , Infecciones por Coxsackievirus/inmunología , Miocarditis/inmunología , Troponina/inmunología , Animales , Infecciones por Coxsackievirus/complicaciones , Infecciones por Coxsackievirus/patología , Masculino , Ratones , Ratones Endogámicos BALB C , Miocarditis/patología , Miocarditis/virología , ARN Viral/análisis , Proteínas Virales/inmunologíaRESUMEN
The proinflammatory cytokine interleukin-17A (IL-17A) is known to mediate antimicrobial activity, but its role during rhinovirus (RV) infections and in asthma needs further investigation. Therefore, we addressed the role of IL-17A during allergic asthma and antiviral immune response in human and murine immunocompetent cells. In this study we found that asthmatic children with a RV infection in their upper airways have upregulated mRNA levels of the antiviral cytokine interferon type I (IFN)-ß and the transcription factor T-box 21 (TBX21) and reduced levels of IL-17A protein in their peripheral blood mononuclear cells (PBMCs). We also found that IL-17A inhibited RV1b replication in infected human lung epithelial cells A549. Furthermore, by using gene array analysis we discovered that targeted deletion of Il17a in murine lung CD4(+) T cells impaired Oas1g mRNA downstream of Ifnß, independently from RV infection. Additionally, in PBMCs of children with a RV infection in their nasalpharyngeal fluid OAS1 gene expression was found downregulated. Finally RV1b inhibited IL-17A production in lung CD4(+) T cells in a setting of experimental asthma. These results indicate that the RV1b inhibits IL-17A in T helper type 17 cells and IL-17A clears RV1b infection in epithelial cells. In both cases IL-17A contributes to fend off RV1b infection by inducing genes downstream of interferon type I pathway.
Asunto(s)
Asma/inmunología , Linfocitos T CD4-Positivos/inmunología , Hipersensibilidad a las Drogas/inmunología , Interleucina-17/inmunología , Infecciones por Picornaviridae/inmunología , Rhinovirus/inmunología , 2',5'-Oligoadenilato Sintetasa/genética , 2',5'-Oligoadenilato Sintetasa/inmunología , Células A549 , Animales , Asma/genética , Asma/virología , Linfocitos T CD4-Positivos/virología , Niño , Preescolar , Hipersensibilidad a las Drogas/genética , Hipersensibilidad a las Drogas/virología , Femenino , Regulación de la Expresión Génica , Humanos , Interferón beta/genética , Interferón beta/inmunología , Interleucina-17/genética , Pulmón/inmunología , Pulmón/virología , Masculino , Ratones , Ratones Noqueados , Ovalbúmina/administración & dosificación , Infecciones por Picornaviridae/genética , Infecciones por Picornaviridae/virología , Cultivo Primario de Células , ARN Mensajero/genética , ARN Mensajero/inmunología , Rhinovirus/crecimiento & desarrollo , Transducción de Señal , Proteínas de Dominio T Box/genética , Proteínas de Dominio T Box/inmunologíaRESUMEN
BACKGROUND: The etiology of chronic urticaria is undefined, but the potential role of infectious agents as one triggering factor has been suggested. The appearance of chronic urticaria in a 16-year old male after a history of a recent parvovirus B19 (B19) infection led us to investigate the association between B19 and chronic urticaria. OBJECTIVES: To investigate whether parvovirus B19 (B19) has a role in chronic urticaria. STUDY DESIGN: We amplified B19 DNA from skin biopsy samples of 36 adult chronic urticaria patients as well as of 22 healthy controls using two sets of separate primers and probe. Circulating IgG and IgM antibodies to B19 were measured from 27 patients and from all controls. RESULTS: B19 DNA was detected in 18 (50%) skin biopsy samples of 36 patients with chronic urticaria. Unexpectedly, also 14 (64%) skin biopsy samples from 22 healthy controls harbored B19 DNA. All 32 persons with positive B19 PCR findings had circulating IgG-class antibodies to B19 major structural protein VP2, but no IgM antibodies. CONCLUSION: Our results show that B19 DNA commonly exists in human skin. Therefore, the association between B19 infection and chronic urticaria remains uncertain. However, these findings raise the question whether the skin may constitute a reservoir for B19.
Asunto(s)
ADN Viral/análisis , Infecciones por Parvoviridae/virología , Parvovirus B19 Humano/aislamiento & purificación , Piel/virología , Urticaria/virología , Adolescente , Adulto , Anciano , Anticuerpos Antivirales/sangre , Enfermedad Crónica , Humanos , Inmunoglobulina G/sangre , Masculino , Persona de Mediana Edad , Parvovirus B19 Humano/inmunología , Reacción en Cadena de la PolimerasaRESUMEN
Quantitative enzyme immunoassays for parainfluenza type 1, 2 and 3 IgG antibodies were developed. Serum specimens were tested at a single dilution of 1:1000 and results expressed in units by the use of a standard curve. The unit values correlated well with titres obtained by testing the same specimens in serial dilutions. All serum pairs with significant titre rises also showed significant rises in unit values. Parainfluenza IgG and IgM serology was evaluated in 66 patients with a proven parainfluenza infection. Diagnostic IgG antibody increases were detected in 70, 69 and 87% of parainfluenza type 1, 2 and 3 infections, respectively. Heterologous titre rises between parainfluenza types 1 and 3 were common. IgM antibodies were detected in 42% of the patients, most commonly in those below two years of age and rarely in adults.
Asunto(s)
Anticuerpos Antivirales/análisis , Técnicas para Inmunoenzimas , Respirovirus/inmunología , Humanos , Inmunoglobulina G/análisis , Inmunoglobulina M/análisis , Virus de la Parainfluenza 1 Humana/inmunología , Virus de la Parainfluenza 2 Humana/inmunología , Virus de la Parainfluenza 3 Humana/inmunología , Infecciones por Paramyxoviridae/diagnóstico , Virología/métodosRESUMEN
Islet-like cell clusters (ICCs) prepared from human fetal pancreases were infected with mumps or coxsackie B3 virus. Double-labeled antibody technique showed that the viruses infected both insulin-secreting and other pancreatic cells and that secretion of immunoreactive insulin into the culture medium of the mumps virus-infected cells had already ceased on day 7. The mumps virus-infected ICC clusters produced virus for 14 days, and the mumps virus antigen was detected in the ICCs through the whole 22-day observation period. The coxsackie B3 virus-infected ICCs contained cells highly positive for viral antigen during the first 2 days after infection, and the infectious virus was detected in the culture medium for 22 days. This in vitro model indicates that mumps and coxsackie B3 viruses infect human fetal pancreatic endocrine cells and are able to alter beta-cell function. Coxsackie B3 virus infection in ICCs is lytical and seems to lead to rapid cell destruction, but long-lasting, restricted mumps virus infection in human fetal pancreatic ICCs offers an interesting model to study the effects of viral infection in the endocrine pancreas and beta cell.
Asunto(s)
Infecciones por Coxsackievirus/fisiopatología , Islotes Pancreáticos/citología , Islotes Pancreáticos/microbiología , Paperas/metabolismo , Páncreas/embriología , Células Cultivadas , Enterovirus Humano B/aislamiento & purificación , Enterovirus Humano B/fisiología , Feto/citología , Técnica del Anticuerpo Fluorescente , Humanos , Insulina/metabolismo , Islotes Pancreáticos/fisiología , Virus de la Parotiditis/aislamiento & purificación , Virus de la Parotiditis/fisiología , Radioinmunoensayo , Factores de Tiempo , Replicación Viral/fisiologíaRESUMEN
D-erythro-Pentos-2-ulose and D-threo-pentos-2-ulose and their 1-13C- and 2-13C-substituted derivatives have been prepared by oxidizing the corresponding natural and 13C-substituted D-aldopentoses (D-arabinose, D-xylose) with cupric acetate, and purifying the products by chromatography on a cation-exchange resin in the calcium or barium form. The equilibrium compositions of the pentos-2-uloses in 2H2O were determined by 13C-n.m.r. spectroscopy (75 MHz) at 25 degrees and 80 degrees. Among the eighteen possible monomeric acyclic, cyclic, and bicyclic forms, the anomeric pairs of the unhydrated aldopyranoses, aldopyranose endocyclic hydrates, aldofuranose endocyclic hydrates, and ketofuranose exocyclic hydrates were identified on the basis of 13C chemical shifts and 13C-1H and 13C-13C spin-coupling constants. 1H-N.m.r. (300, 500, and 620 MHz) and 13C-n.m.r. (75 MHz) spectroscopic data in one and two dimensions (DQF-COSY, homonuclear 2D-J) were used to evaluate the conformational properties of the cyclic structures. The unhydrated pyranoses are highly conformationally homogeneous; the erythro and threo isomers prefer 1C4 and 4C1 conformations, respectively. D-threo-Pentos-2-ulopyranose hydrate prefers the 4C1 conformation whereas the erythro isomers exists in both the 4C1 and 1C4 conformations. The furanoid forms favor structures having quasi-axial anomeric hydroxyl groups and quasi-equatorial exocyclic hydroxymethyl or dihydroxymethyl groups.
Asunto(s)
Arabinosa/análogos & derivados , Cetosas , Xilosa/análogos & derivados , Arabinosa/síntesis química , Arabinosa/química , Conformación de Carbohidratos , Isótopos de Carbono , Cromatografía de Gases , Hidrógeno , Espectroscopía de Resonancia Magnética , Estructura Molecular , Xilosa/síntesis química , Xilosa/químicaRESUMEN
The pentuloses D-erythro-2-pentulose (1) and D-threo-2-pentulose (2) and their 1-13C- and 2-13C-substituted derivatives were prepared by hydrogenating the corresponding isotopically normal and 13C-substituted D-pentos-2-uloses with a Pd-carbon catalyst. The threo isomer and its labeled derivatives were alternatively prepared from isotopically normal and 13C-substituted D-xyloses with immobilized D-xylose (D-glucose) isomerase (E.C.5.3.1.5). The equilibrium compositions of 1 and 2 (furanose anomers and acyclic keto forms) in 2H2O were determined from 13C-n.m.r. spectra (75 MHz) of the 2-13C-labeled derivatives. The conformational properties of the cyclic and acyclic forms in 2H2O were assessed with the use of 1H-1H, 13C-1H, and 13C-13C spin-coupling constants obtained from 1H-n.m.r. (620 MHz) and 13C-n.m.r. (75 MHz) spectra. Compared with the structurally related aldotetrofuranoses the 2-pentulofuranoses more strongly prefer conformations in which the anomeric hydroxyl group is oriented quasi-axially. The strongly dipolarized carbonyl group in the acyclic keto forms of 1 and 2 appears to stabilize chain conformations having O-1 and O-3 eclipsed with the carbonyl oxygen.
Asunto(s)
Pentosas/síntesis química , Xilulosa/síntesis química , Conformación de Carbohidratos , Isótopos de Carbono , Espectroscopía de Resonancia Magnética , Pentosas/química , Xilulosa/químicaRESUMEN
The tautomeric compositions of D-erythro-2-pentulose (D-ribulose) and D-threo-2-pentulose (D-xylulose) in aqueous solution have been studied by 13C-n.m.r. spectroscopy at various temperatures using 2-13C-substituted compounds. The alpha-furanose, beta-furanose, and acyclic carbonyl (keto) forms were detected at all temperatures, whereas the acyclic hydrate (gem-diol) form was not observed. The percentage of keto form increased with increasing temperature, at the expense of the furanose forms. Thermodynamic (delta G0, delta H0, delta S0) and kinetic parameters for the interconversion of alpha- and beta-furanoses with the acyclic carbonyl form were determined and compared with those determined under similar conditions for the structurally-related aldotetrofuranoses. The ring-opening rate constant (kopen) measured by 13C saturation-transfer n.m.r. spectroscopy in 50mM sodium acetate (pH 4.0) at 55 degrees were as follows: beta-threofuranose (0.65 s-1) greater than alpha-erythrofuranose (0.51 s-1) greater than beta-erythrofuranose (0.37 s-1) approximately beta-threo-2-pentulofuranose (0.35 s-1) greater than alpha-threofuranose (0.25 s-1) greater than alpha-threo-2-pentulofuranose (0.20 s-1) approximately alpha-erythro-2-pentulofuranose (0.18 s-1) approximately beta-erythro-2-pentulofuranose (0.18 s-1). Within each structural type the pentulofuranose anomer having O-2 and O-3 cis (O-1 and O-2 cis in aldotetrofuranoses) opens faster than, or at a similar rate to, the alternative anomer having these oxygen atoms trans. Ring-closing rate constants (kclose), calculated from kopen and Keq, decrease in the order beta-erythrofuranose (15 s-1) greater than beta-threofuranose (12 s-1) greater than alpha-erythrofuranose (9.9 s-1) greater than alpha-threofuranose (6.2 s-1) greater than beta-threo-2-pentulofuranose (0.71 s-1) greater than alpha-erythro-2-pentulofuranose (0.38 s-1) greater than alpha-threo-2-pentulofuranose (0.13 s-1) approximately beta-erythro-2-pentulofuranose (0.13 s-1). Replacement of H-1 in aldotetrofuranoses by a hydroxymethyl group (i.e., conversion to 2-pentuloses) significantly decreases the ring-opening and ring-closing rate constants of furanose anomerization.
Asunto(s)
Pentosas/química , Isomerismo , Cinética , Espectroscopía de Resonancia Magnética , Estructura Molecular , Termodinámica , Xilulosa/químicaRESUMEN
A 4-O-methylglucuronoxylan was converted into a hexenuronoxylan at high temperature and alkalinity similar to the conditions used during kraft pulping. The hexenuronoxylan was hydrolysed with enzymes, and acidic xylooligosaccharides were separated from the hydrolysate by anion-exchange and size-exclusion chromatography. The primary structure of the two main hexenuronic acid-substituted xylooligosaccharides (a tetramer and a pentamer) was determined by two-dimensional 1H and 13C NMR spectroscopy. The 4-deoxy-hexenuronic acid is not stable under the acid hydrolysis step of conventional carbohydrate analysis. Here, we have identified the acidic degradation products of 4-deoxy-hexenuronic acid by NMR spectroscopy. Two degradation pathways were observed, both resulting in a furan derivative.