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1.
Br J Anaesth ; 107(4): 533-9, 2011 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-21659406

RESUMEN

BACKGROUND: Simulation has been shown to be effective in teaching complex emergency procedural skills. However, the retention of these skills for a period of up to 1 yr has not been studied. We aimed to investigate the 6 month and 1 yr retention of the complex procedural skill of cricothyroidotomy in attending anaesthetists using a high-fidelity-simulated cannot intubate, cannot ventilate (CICV) scenario. METHODS: Thirty-eight attending anaesthetists participated individually in a high-fidelity-simulated CICV scenario (pretest) that required a cricothyroidotomy for definitive airway management. Immediately after a debriefing and structured teaching session on cricothyroidotomy insertion, subjects managed a second identical CICV scenario (post-test). Each anaesthetist was randomized to either a '6 month retention' or a '12 month retention' group. No further teaching occurred. At their respective retention times, each anaesthetist managed a third identical CICV scenario (retention post-test). Two blinded experts independently rated videos of all performances in a random order, using a specific checklist (CL) score, a global-rating scale (GRS) score, and procedural time (PT). RESULTS: Subjects from both groups improved on their cricothyroidotomy skill performances from pretest to immediate post-test and from pretest to retention post-test, irrespective of the retention interval; CL mean (sd) 8.00 (2.39) vs 8.88 (1.53), P=0.49; GRS 28.00 (7.80) vs 31.25 (5.31), P=0.25; PT 102.83 (63.81) s vs 106.88 (36.68) s, P=0.73. CONCLUSIONS: After a single simulation training session, improvements in cricothyroidotomy skills are retained for at least 1 yr. These findings suggest that high-fidelity simulation training, along with practice and feedback, can be used to maintain complex procedural skills for at least 1 yr.


Asunto(s)
Manejo de la Vía Aérea/métodos , Anestesia , Anestesiología/educación , Competencia Clínica , Servicios Médicos de Urgencia/métodos , Complicaciones Intraoperatorias/terapia , Maniquíes , Cartílago Cricoides/cirugía , Humanos , Aprendizaje , Variaciones Dependientes del Observador , Tamaño de la Muestra , Método Simple Ciego , Tiroidectomía , Factores de Tiempo
2.
J Appl Microbiol ; 108(3): 908-916, 2010 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-19709336

RESUMEN

AIMS: To isolate and characterize an antagonist for use as probiotic agent in the biocontrol of Staphylococcus aureus. METHODS AND RESULTS: Bacteria that exhibited antimicrobial activity against Gram-positive bacteria including Staph. aureus were isolated from 12 healthy women, with Staphylococcus hominis MBBL 2-9 showing the strongest activity. The bacteriocin produced by Staph. hominis MBBL 2-9 was purified by 60% ammonium sulfate saturation, ultrafiltration, HLB cartridge and reverse-phase HPLC. The molecular weight was estimated as 2038.2 Da by MALDI-TOF mass spectrometry. The antagonist survived up to 2 h in artificial gastric juice (pH 2.5) and grew in the presence of 1% porcine bile extract. In addition, Staph. hominis MBBL 2-9 adhered effectively to HT-29 epithelial cell line. CONCLUSION: Staphylococcus hominis MBBL 2-9 exhibited desirable probiotic traits such as acid tolerance, bile resistance and adherence to epithelial cell line. The bacterium also produced a bacteriocin with unique molecular weight and high antimicrobial activity similar to traditional antibiotics. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is the first report of a bacteriocin-producing Staph. hominis MBBL 2-9 that has potential for use as a probiotic agent against Staph. aureus.


Asunto(s)
Probióticos , Infecciones Estafilocócicas/prevención & control , Staphylococcus aureus/aislamiento & purificación , Staphylococcus hominis/fisiología , Vagina/microbiología , Adulto , Antibacterianos/farmacología , Antibiosis , Adhesión Bacteriana , Bacteriocinas/biosíntesis , Bacteriocinas/aislamiento & purificación , Línea Celular Tumoral , Femenino , Humanos , Pruebas de Sensibilidad Microbiana , Viabilidad Microbiana , Infecciones Estafilocócicas/microbiología , Staphylococcus aureus/efectos de los fármacos , Staphylococcus hominis/metabolismo
3.
J Environ Qual ; 39(5): 1807-12, 2010.
Artículo en Inglés | MEDLINE | ID: mdl-21043286

RESUMEN

About 80% of dairy cattle N intake is excreted in urine and feces. Urinary-N is about 75% urea, whereas fecal-N is mostly organic. Urinary-N (urea) can only be volatilized when it is hydrolyzed to ammonia (NH3) in a process catalyzed by urease, which is predominantly found in feces. Minimizing contact between urine and feces may be an effective approach to reducing urea hydrolysis and subsequent NH3 emissions. Previous studies have reported 5 to 99% NH3 emissions mitigation within barns from separation of feces and urine. The objective ofthis study was to compare NH3 emissions mitigation via separation of urine and feces in postcollection storage to a conventional scrape manure handling method where urine and feces are comingled. Laboratory scale studies were conducted to evaluate NH3 emissions from simulated postcollection storag of three waste streams: (i) idealistically separated feces and urine (no contact between urine and feces), (ii) realistically separated urine and feces (limited contact of urine and feces), and (iii) conventionally scraped manure (control). From the results of these studies, NH3 losses ranking in descending order was as follows: aggregate of realistically separated waste streams (3375.9 +/- 54.8 mg), aggregate of idealistically separated urine and feces (3047.0 +/- 738.0 mg), and scrape manure (2034.0 +/- 106.5 mg), respectively. Therefore, on the basis of these results, the extra effort of separating the waste streams would not enhance mitigation of NH3 losses from postcollection storage of the separated waste streams compared to the conventional scrape manure collection system.


Asunto(s)
Amoníaco/análisis , Industria Lechera , Animales , Bovinos , Heces , Orina
4.
Br J Anaesth ; 103(4): 570-5, 2009 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-19689979

RESUMEN

BACKGROUND: Retention of skills and knowledge after neonatal resuscitation courses (NRP) is known to be problematic. The use of cognitive aids is mandatory in industries such as aviation, to avoid dependence on memory when decision-making in critical situations. We aimed to prospectively investigate the effect of a cognitive aid on the performance of simulated neonatal resuscitation. METHODS: Thirty-two anaesthesia residents were recruited. The intervention group had a poster detailing the NRP algorithm and the control group did not. Video recordings of each of the performances were analysed using a previously validated checklist by a peer, an expert anaesthetist, and an expert neonatologist. RESULTS: The median (IQR) checklist score in the control group [18.2 (15.0-20.5)] was not significantly different from that in the intervention group [20.3 (18.3-21.3)] (P=0.08). When evaluated by the neonatologist, none of the subjects correctly performed all life-saving interventions necessary to pass the checklist. A minority of the intervention group used the cognitive aid frequently. CONCLUSIONS: Retention of skills after NRP training is poor. The infrequent use of the cognitive aid may be the reason that it did not improve performance. Further research is required to investigate whether cognitive aids can be useful if their use is incorporated into the NRP training.


Asunto(s)
Algoritmos , Reanimación Cardiopulmonar/educación , Competencia Clínica , Reanimación Cardiopulmonar/normas , Protocolos Clínicos , Técnicas de Apoyo para la Decisión , Educación Médica Continua , Femenino , Humanos , Recién Nacido , Cuidado Intensivo Neonatal/métodos , Cuidado Intensivo Neonatal/normas , Masculino , Ontario , Estudios Prospectivos , Retención en Psicología , Método Simple Ciego
5.
J Environ Qual ; 38(2): 647-53, 2009.
Artículo en Inglés | MEDLINE | ID: mdl-19244485

RESUMEN

Strong acid solutions have been widely used in acid traps to determine concentrations of ammonia in ambient air or exhaust air stream. A literature survey indicates the method has a long history and a wide variation in use. Through a series of studies, this paper examines several factors including volume of the acid, depth of the acid, and airflow rate; that might affect the efficiency of sulfuric acid traps and recommends steps researchers and other users may take to ensure reliable results from this method. The results from these series of studies indicate: (i) an inverse relationship between the efficiency of the acid traps and the amount of ammonia to be trapped even when the capacity of the acid trap is excessively greater than the maximum theoretical stoichiometric capacity needed to dissolve all of the ammonia, (ii) for the same volume of acid, the efficiency of the acid trap increased with the acid depth but overall, the efficiency at any given acid depth decreased as the amount of ammonia through the trap increased, and (iii) at the two airflow rates examined in this study (0.5 and 1.0 L/min) the efficiency of the acid traps decreased at similar rates as the concentration of ammonia in the sample air increased but the efficiency of the trap was significantly higher at the lower airflow rate. To obtain reliable measurements from this method, therefore, multi-point calibrations within the entire range of target measurements is recommended to provide accurate corrections of the measurements.


Asunto(s)
Aire/análisis , Amoníaco/análisis , Ácidos Sulfúricos/química
6.
Oncogene ; 37(3): 377-388, 2018 01 18.
Artículo en Inglés | MEDLINE | ID: mdl-28945228

RESUMEN

Hyperactivation of phosphatidylinositol 3-kinase (PI3K) pathway occurs frequently in head and neck squamous cell carcinoma (HNSCC). However, clinical outcomes of targeting the PI3K pathway have been underwhelming. In present study, we investigated the resistant mechanisms and potential combination therapeutic strategy to overcome adaptive resistance to PI3K inhibitor in HNSCC. Treatment of NVP-BKM120, a pan-PI3K inhibitor, led to upregulation of interleukin-6 (IL-6) and subsequent activation of either extracellular signal-regulated kinase (ERK) or signal transducers and activators of transcription 3 (STAT3), causing modest antitumor effects on the growth of HNSCC cells. Blockade of autocrine IL-6 signaling with siRNA or neutralizing antibody for IL-6 receptor (IL-6R) completely abolished NVP-BKM120-induced activation of ERK and STAT3 as well as expression of c-Myc oncogene, which resulted in enhanced sensitivity to NVP-BKM120. Moreover, when compared with a pharmacologic inhibitor or silencing of STAT3, trametinib, a MEK inhibitor, in combination with NVP-BKM120 yielded more potent anti-proliferative effects by inhibiting S phase transition, arresting cells at G0/G1 phase, and downregulating IL-6 and c-Myc expression. Furthermore, as compared with either agent alone, combination of NVP-BKM120 with trametinib or tocilizumab, a humanized anti-IL-6R antibody, significantly suppressed tumor growth in NVP-BKM120-resistant patient-derived tumor xenograft (PDTX) models, which was also confirmed in PDTX-derived cell lines. Collectively, these results suggested that IL-6/ERK signaling is closely involved in adaptive resistance of NVP-BKM120 in HNSCC cells, providing a rationale for a novel combination therapy to overcome resistance to PI3K inhibitors.


Asunto(s)
Aminopiridinas/farmacología , Protocolos de Quimioterapia Combinada Antineoplásica/farmacología , Carcinoma de Células Escamosas/tratamiento farmacológico , Resistencia a Antineoplásicos , Neoplasias de Cabeza y Cuello/tratamiento farmacológico , Interleucina-6/metabolismo , Morfolinas/farmacología , Inhibidores de las Quinasa Fosfoinosítidos-3 , Inhibidores de Proteínas Quinasas/farmacología , Aminopiridinas/uso terapéutico , Animales , Anticuerpos Monoclonales Humanizados/farmacología , Anticuerpos Monoclonales Humanizados/uso terapéutico , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Comunicación Autocrina/efectos de los fármacos , Carcinoma de Células Escamosas/patología , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Quinasas MAP Reguladas por Señal Extracelular/metabolismo , Femenino , Neoplasias de Cabeza y Cuello/patología , Humanos , Interleucina-6/genética , Sistema de Señalización de MAP Quinasas/efectos de los fármacos , Ratones , Ratones Endogámicos NOD , Morfolinas/uso terapéutico , Fosfatidilinositol 3-Quinasas/metabolismo , Inhibidores de Proteínas Quinasas/uso terapéutico , Piridonas/farmacología , Piridonas/uso terapéutico , Pirimidinonas/farmacología , Pirimidinonas/uso terapéutico , ARN Interferente Pequeño/metabolismo , Receptores de Interleucina-6/antagonistas & inhibidores , Receptores de Interleucina-6/metabolismo , Factor de Transcripción STAT3/metabolismo , Carcinoma de Células Escamosas de Cabeza y Cuello , Ensayos Antitumor por Modelo de Xenoinjerto
7.
Oncogene ; 36(39): 5512-5521, 2017 09 28.
Artículo en Inglés | MEDLINE | ID: mdl-28534506

RESUMEN

Lysine-specific demethylase 1 (LSD1), which has been considered as a potential therapeutic target in human cancer, has been known to regulate many biological functions through its non-histone substrates. Although LSD1-induced hypoxia-inducible factor alpha (HIF1α) demethylation has recently been proposed, the effect of LSD1 on the relationship between HIF1α post-translational modifications (PTMs) and HIF1α-induced tumor angiogenesis remains to be elucidated. Here, we identify a new methylation site of the HIF1α protein antagonized by LSD1 and the interplay between HIF1α protein methylation and other PTMs in regulating tumor angiogenesis. LSD1 demethylates HIF1α at lysine (K) 391, which protects HIF1α against ubiquitin-mediated protein degradation. LSD1 also directly suppresses PHD2-induced HIF1α hydroxylation, which has a mutually dependent interplay with Set9-mediated HIF1α methylation. Moreover, the HIF1α acetylation that occurs in a HIF1α methylation-dependent manner is inhibited by the LSD1/NuRD complex. HIF1α stabilized by LSD1 cooperates with CBP and MTA1 to enhance vascular endothelial growth factor (VEGF)-induced tumor angiogenesis. Thus, LSD1 is a key regulator of HIF1α/VEGF-mediated tumor angiogenesis by antagonizing the crosstalk between PTMs involving HIF1α protein degradation.


Asunto(s)
Neoplasias de la Mama/irrigación sanguínea , Histona Demetilasas/metabolismo , Subunidad alfa del Factor 1 Inducible por Hipoxia/metabolismo , Animales , Neoplasias de la Mama/genética , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Línea Celular Tumoral , Femenino , Células HEK293 , Xenoinjertos , Histona Demetilasas/genética , Células Endoteliales de la Vena Umbilical Humana , Humanos , Subunidad alfa del Factor 1 Inducible por Hipoxia/genética , Ratones , Ratones Endogámicos NOD , Ratones SCID , Neovascularización Patológica/metabolismo , Neovascularización Patológica/patología , Transcripción Genética , Transfección , Ubiquitina/metabolismo
8.
J Virol Methods ; 102(1-2): 53-9, 2002 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-11879692

RESUMEN

A total of 360 type A swine influenza virus-positive samples including cell culture isolates, nasal swabs or lung tissues along with 30 virus-negative samples were tested for the detection and subtyping of H1N1, H1N2 or H3N2 by two multiplex reverse transcription (RT)-PCR assays. The positive samples had been collected between 1999 and 2001 from pigs with respiratory diseases, and type A influenza virus was isolated and subtyped by hemagglutination inhibition (HI) test at the Minnesota Veterinary Diagnostic Laboratory (MVDL). Two multiplex RT-PCR assays specific for H1 and H3, and N1 and N2 were developed. RT-PCR products with unique sizes characteristic of each subtype of influenza A virus were sequenced, and the sequences were demonstrated to be specific for H1N1, H1N2 or H3N2. Genomic RNAs or DNAs from 12 common swine pathogens other than type A influenza viruses were not amplified when the PCR assays were performed with these primer sets. Positive amplification reaction could be visualized with RNA extracted from up to 10(-5) dilution of each reference virus with original infectivity titer of 10(5) TCID(50)/ml. Of the 360 samples tested, swine influenza virus H1N1, H1N2 and H3N2 were identified in 200, 13 and 139 samples, respectively. The remaining eight samples were positive for both H1N1 and H3N2 viruses. The results of multiplex RT-PCR were 100% in agreement with those of virus isolation. These results demonstrate the usefulness of multiplex RT-PCR for detection and identification of influenza A virus subtypes. The results also indicate an increased occurrence of H1N2 in US swine population.


Asunto(s)
Subtipo H1N1 del Virus de la Influenza A , Subtipo H3N2 del Virus de la Influenza A , Virus de la Influenza A/aislamiento & purificación , Infecciones por Orthomyxoviridae/veterinaria , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Enfermedades de los Porcinos/virología , Animales , Línea Celular , Virus de la Influenza A/clasificación , Virus de la Influenza A/genética , Infecciones por Orthomyxoviridae/virología , Porcinos
9.
Vet Microbiol ; 17(4): 315-22, 1988 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-2847396

RESUMEN

A radial immunodiffusion enzyme assay (RIDEA) was developed for detection and quantitation of antibodies to equine herpes virus-1 (EHV-1) in horse sera. The detection and quantitation of EHV-1 antibody levels were based on the diameter of the radial diffusion zone of specific antibody in each serum sample reacting with EHV-1 antigen. The circular zone was made visible using peroxidase-conjugated rabbit anti-horse immunoglobulin G and a substrate containing hydrogen peroxide. The results of the RIDEA were compared with those of virus neutralization (VN) and enzyme-linked immunosorbent assay (ELISA) and found to be highly correlated. The relative sensitivity and specificity (percentage of agreement with VN test) were found to be 98.2 and 92.5%, respectively. Because the test procedure is relatively easy to perform, the RIDEA could be used as a field test to detect antibodies to EHV-1 in horses.


Asunto(s)
Anticuerpos Antivirales/análisis , Herpesviridae/inmunología , Herpesvirus Équido 1/inmunología , Caballos/inmunología , Inmunodifusión , Técnicas para Inmunoenzimas , Animales , Ensayo de Inmunoadsorción Enzimática , Pruebas de Neutralización , Valor Predictivo de las Pruebas
10.
Vet Microbiol ; 55(1-4): 303-7, 1997 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-9220626

RESUMEN

IgG and IgM antibody responses were examined by an indirect fluorescent antibody method in pigs following inoculation with different porcine reproductive and respiratory syndrome virus (PRRSV) isolates or a vaccine virus. Viremia was also examined in the pigs. The IgG antibody was first detected between 9 and 14 days post inoculation (PI) and maintained high titers for at least 7 weeks PI. No change in IgG antibody titers was observed when the pigs were reinoculated with PRRSV 35 days PI. IgM antibody was detected between 5 and 28 days PI in the pigs. Reinoculation at 35 days PI caused a short term rise of IgM antibody. Virus was isolated from sera collected between 2 and 21 days PI. The IgM antibody was detected regularly in sera collected during viremia and up to 1-2 weeks after the viremic periods. These results suggest that pigs with detectable IgM antibody are probably pigs with recent infection and that routine testing of IgM antibody in purchased breeding pigs from seropositive farms may be useful in identification of pigs with recent infection.


Asunto(s)
Anticuerpos Antivirales/sangre , Inmunoglobulina M/sangre , Síndrome Respiratorio y de la Reproducción Porcina/inmunología , Virus del Síndrome Respiratorio y Reproductivo Porcino/inmunología , Animales , Técnica del Anticuerpo Fluorescente Indirecta , Inmunoglobulina G/sangre , Síndrome Respiratorio y de la Reproducción Porcina/sangre , Síndrome Respiratorio y de la Reproducción Porcina/diagnóstico , Porcinos , Factores de Tiempo , Viremia/sangre , Viremia/diagnóstico , Viremia/inmunología
11.
Vet Microbiol ; 10(3): 209-18, 1985 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-4002607

RESUMEN

Fourteen different adjuvants, given either in single or combined form with another compound were compared in guinea pigs for their ability to potentiate humoral immunity to porcine parvovirus (PPV) antigen after 2 vaccinations. Two injections were given, the second 3 weeks following the initial vaccination. Antibody concentrations to PPV in sera from injected animals were measured over a 5-week period by the hemagglutination inhibition test. At the conclusion of the experiment, guinea pigs injected with the following adjuvants and PPV antigen: CP-20 961 (Avridin), 50% aluminum hydroxide gel, ethylene maleic anhydride (EMA), oil and water emulsion (O/W) and dimethyl-dioctadecyl-ammonium bromide (DDA) immunologically responded with high geometric mean HI titers (380, 224 and 427, 602, 512, 1202 respectively), whereas guinea pigs receiving Emulsan, sodium dodecyl sulfate (SDS), L-121, combinations of Emulsan/aluminum hydroxide, SDS/aluminum hydroxide and B. pertussis/aluminum hydroxide responded with low mean titers (54, 64, 18, 27, 11, 64, 14, 20 respectively). Guinea pigs injected with antigen without adjuvant responded weakly with geometric mean titers of 3.3 and 16 for the 2 groups tested. Prior to booster injection, guinea pigs immunized with 13 of the preparations had low (less than 4) or undetectable antibody titers. Antibody titers from guinea pigs receiving DDA adjuvant continued to rise throughout the duration of the experiment and at the conclusion had the highest mean titers of the groups tested (1202). The 2 groups immunized with 50% aluminum hydroxide gel had high mean titers (224, 427), but in both instances there was a wide range of titers within a group evidenced by high standard deviations. In contrast, guinea pigs receiving either DDA, CP-20 961, O/W or EMA had antibody titers within a narrow range and small standard deviation. The significance of aluminum hydroxide gel concentration on immunogenicity is discussed.


Asunto(s)
Adyuvantes Inmunológicos/inmunología , Formación de Anticuerpos , Parvoviridae/inmunología , Vacunas Virales/inmunología , Hidróxido de Aluminio/inmunología , Animales , Antígenos Virales/inmunología , Cobayas , Pruebas de Inhibición de Hemaglutinación , Vacunación
12.
Vet Microbiol ; 25(2-3): 177-92, 1990 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-2126409

RESUMEN

An enzyme-linked immunosorbent assay (ELISA) was evaluated for detection of antibodies (Ab) against Mycoplasma hyopneumoniae and M. flocculare in sera from swine experimentally infected with these agents. In addition, the ELISA was compared with the complement fixation test (CFT), and radial immunodiffusion enzyme assay (RIDEA) for the demonstration of Ab against M. hyopneumoniae. Twenty two 6-week-old swine from a respiratory disease-free herd were divided into five groups. Two or three pigs from each of the four groups were inoculated, respectively, with M. hyopneumoniae or with M. flocculare while two pigs in each group were contact exposed to the inoculated penmates. A fifth group, consisting of three pigs, served as inoculated controls. Pigs inoculated with M. hyopneumoniae began coughing 13 days post inoculation (PI). Antibodies were first detected 2 weeks PI with the CFT, 3 weeks PI with the ELISA, and 5 weeks PI with the RIDEA. With the ELISA and RIDEA, Ab were still detectable one year PI at a very low level. With the CFT, Ab were not detectable in sera from any swine beyond 5 months PI. At necropsy 1 year PI, no lesions were detected in lungs of any of the animals nor were mycoplasmas detected. M. flocculare inoculated or contact-exposed pigs never evidenced clinical signs. Antibodies against M. flocculare were first detected 5 to 12 weeks PI with CFT, and 6 to 12 weeks PI with the ELISA. Peak optical density (OD) values obtained in the ELISA with M. flocculare Ab were as high as the values obtained with peak M. hyopneumoniae Ab titers. Levels of Ab against M. flocculare were at relatively higher OD at 1 year PI than Ab against M. hyopneumoniae. Sera with high levels of Ab against M. flocculare cross-reacted slightly with M. hyopneumoniae antigen in immunoblotting and ELISA.


Asunto(s)
Anticuerpos Antibacterianos/sangre , Ensayo de Inmunoadsorción Enzimática , Mycoplasma/inmunología , Neumonía Porcina por Mycoplasma/veterinaria , Enfermedades de los Porcinos/inmunología , Animales , Pruebas de Fijación del Complemento , Reacciones Cruzadas , Electroforesis en Gel de Poliacrilamida , Estudios de Evaluación como Asunto , Immunoblotting , Inmunodifusión , Neumonía Porcina por Mycoplasma/diagnóstico , Neumonía Porcina por Mycoplasma/inmunología , Valor Predictivo de las Pruebas , Porcinos , Enfermedades de los Porcinos/diagnóstico
13.
Vet Microbiol ; 15(1-2): 19-29, 1987 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-2830705

RESUMEN

The pathogenic properties of a skin isolate of porcine parvovirus (PPV), designated Kresse isolate, were compared with NADL-8 isolate, a prototype isolate of PPV, by in utero inoculation of mid-term and late-term gestation swine fetuses. Fetuses from pregnant sows of mid-gestation were inoculated with either NADL-8 or Kresse virus. Both isolates were highly pathogenic to mid-gestation fetuses. In contrast, dramatic differences in pathogenicity between these 2 isolates were observed in fetuses inoculated late in gestation. Such fetuses from each of 4 sows were inoculated with NADL-8 or Kresse virus isolate and sacrificed at 10, 18, 21, or 23 days postinoculation (PI). NADL-8-inoculated fetuses were grossly normal. The pathogenic effects of Kresse isolate were evident by gross pathology in fetuses collected at 18, 21, and 23 days PI, but not at 10 days PI. Hemagglutination (HA) and fluorescent antibody (FA) methods were used to identify virus in various tissues of late-gestation fetuses collected at 10 and 21 days PI. At 10 days PI, HA antigens were detected only in livers of NADL-8-inoculated fetuses, but in all tissues examined of Kresse-inoculated fetuses, including the brain. PPV specific fluorescence was demonstrated in tissues of fetuses inoculated with NADL-8 and Kresse virus. The major difference was that virus antigen was found in the brains of fetuses inoculated with Kresse virus, but not in NADL-8 infected fetuses. At 21 days PI, HA antigen was not detected in any of the tissues of fetuses inoculated with NADL-8 virus, with PPV specific fluorescence by FA being found only in the kidney. However, fetuses inoculated with Kresse virus displayed HA antigen in liver and PPV-specific fluorescence in all tissues tested including the brain. Both isolates induced similar antibody responses, 1:128 to 1:256 at 10 days and 1:512 to 1:1024 at 21 days PI. In addition, immunoglobulin G (IgG) deposits were demonstrated in kidneys and skin of fetuses inoculated with Kresse virus and IgM in brain, but not in tissues from fetuses inoculated with NADL-8 virus.


Asunto(s)
Enfermedades Fetales/veterinaria , Infecciones por Parvoviridae/veterinaria , Parvoviridae/patogenicidad , Enfermedades de los Porcinos/microbiología , Animales , Antígenos Virales/análisis , Femenino , Enfermedades Fetales/microbiología , Enfermedades Fetales/patología , Técnica del Anticuerpo Fluorescente , Pruebas de Hemaglutinación , Parvoviridae/inmunología , Infecciones por Parvoviridae/microbiología , Infecciones por Parvoviridae/patología , Embarazo , Complicaciones Infecciosas del Embarazo/microbiología , Complicaciones Infecciosas del Embarazo/patología , Complicaciones Infecciosas del Embarazo/veterinaria , Piel/microbiología , Porcinos , Enfermedades de los Porcinos/patología
14.
Vet Microbiol ; 9(1): 27-33, 1984 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-6719818

RESUMEN

Antibody responses were compared in guinea-pigs, rabbits and pigs following vaccination with inactivated porcine parvovirus (PPV) vaccines. Mean PPV hemagglutination inhibition (HI) antibody titers of 52, 56 and 36 at 1 week after first vaccination and 896, 640 and 512 at 2 weeks after second vaccination were detected in guinea-pigs, rabbits and pigs, respectively. PPV vaccines prepared with greater concentrations of virus, as determined by hemagglutination (HA) units, and of aluminum hydroxide gel adjuvant, induced higher HI antibody titers in guinea-pigs. Optimal concentrations for inducing consistently high antibody titers consisted of vaccine virus with a HA titer of 256/0.1 ml and gel adjuvant at a final concentration of 50%. A second vaccination at 4 weeks compared to 2 or 3 weeks after first vaccination resulted in higher mean HI titers. These data provide preliminary information on the use of guinea-pigs or rabbits as laboratory animal models for testing the potency of PPV vaccines.


Asunto(s)
Anticuerpos Antivirales/biosíntesis , Parvoviridae/inmunología , Porcinos/inmunología , Vacunación/veterinaria , Vacunas Virales/inmunología , Animales , Cobayas/inmunología , Pruebas de Inhibición de Hemaglutinación , Hemaglutininas Virales/análisis , Inmunización Secundaria , Conejos/inmunología , Vacunas Atenuadas/inmunología
15.
Comp Immunol Microbiol Infect Dis ; 10(3-4): 167-71, 1987.
Artículo en Inglés | MEDLINE | ID: mdl-2827946

RESUMEN

The serum-neutralization test (SN), enzyme-linked immunosorbent assay (ELISA) and the radial immunodiffusion enzyme assay (RIDEA) were compared for the detection of pseudorabies (PRV) antibodies in swine sera. A total of 1285 serum samples were tested. All three tests were considered useful in determining the PRV antibody status of swine on a herd basis, but available evidence supports the continued use of SN as the definitive test because of possible false positive reactions associated with ELISA and RIDEA.


Asunto(s)
Anticuerpos Antivirales/análisis , Ensayo de Inmunoadsorción Enzimática , Herpesvirus Suido 1/inmunología , Inmunodifusión , Pruebas de Neutralización , Porcinos/inmunología , Animales , Reacciones Falso Positivas
16.
Vet Immunol Immunopathol ; 22(2): 175-86, 1989 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-2815578

RESUMEN

Three experiments were performed to evaluate the inflammatory response, the antibody response and protection from experimental challenge of various Actinobacillus pleuropneumoniae serotype 5 (Ap5) vaccines in swine. In the first experiment, subcutaneous injections of either a water-in-oil (W/O) emulsion or Freund's complete adjuvant (FCA) caused lesions at the site of injection, while intraperitoneal injection of the W/O emulsion caused no lesions. In the second experiment, intraperitoneal (IP) injection of a W/O emulsion containing unwashed Ap5 cells (6-h culture) and/or supernates from a 24-h culture resulted in severe peritoneal lesions, while W/O emulsion containing PBS-washed Ap5 cells resulted in minimal peritoneal lesions. Ap5 alone or W/O alone failed to cause peritoneal lesions. The third experiment compared the antibody response and protection from challenge of pigs immunized with either 6-h PBS-washed Ap5 cells emulsified in oil - IP, 6-hour Ap5 cells adjuvanted with dimethyl diodacyl ammonium bromide - IP, Ap5 antigen alone - IP, a commercial vaccine - subcutaneously or saline - IP. All groups, except the saline-treated group, responded with high antibody titers to Ap5 2 weeks following vaccination; however, titers from the W/O plus antigen group were significantly higher than the three other groups (P less than 0.05). Following intranasal challenge with Ap5, all animals responded with increased antibody titers. All pigs were euthanized 10 days after challenge and evaluated for pneumonia and the lungs cultured for bacteria. The lungs of all pigs, excepting the W/O plus antigen group, contained pneumonic lesions and A. pleuropneumoniae was cultured from these lesions. These results, along with results from other groups, suggest that intraperitoneal immunization using oil-adjuvanted vaccine may be an effective method for protecting pigs from pneumonia due to A. pleuropneumoniae. Its efficacy may be due to stimulation of local respiratory mucosal immunity.


Asunto(s)
Infecciones por Actinobacillus/veterinaria , Actinobacillus/inmunología , Adyuvantes Inmunológicos/administración & dosificación , Anticuerpos Antibacterianos/biosíntesis , Vacunas Bacterianas/inmunología , Enfermedades de los Porcinos/prevención & control , Actinobacillus/aislamiento & purificación , Infecciones por Actinobacillus/prevención & control , Animales , Antígenos Bacterianos/inmunología , Vacunas Bacterianas/administración & dosificación , Ensayo de Inmunoadsorción Enzimática/veterinaria , Inmunidad Activa , Inyecciones Intraperitoneales , Porcinos , Enfermedades de los Porcinos/inmunología
17.
J Vet Diagn Invest ; 6(3): 293-6, 1994 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-7948197

RESUMEN

Porcine reproductive and respiratory syndrome virus (PRRSV) MN-1b strain open reading frame 4 (ORF4) has been cloned, sequenced, and expressed in Escherichia coli. The homologies of nucleotide and amino acid sequences between MN-1b (US isolate) and LV (European isolate) are 69% and 64%, respectively. The data also showed that ORF4 of MN-1b is 36 bases shorter than that of LV. Western blot analysis of expressed recombinant ORF4 protein reacted with 65% (26/40) of PRRSV-infected pig sera tested. These results demonstrated that ORF4 of PRRSV may not be a well-conserved region.


Asunto(s)
Regulación Viral de la Expresión Génica , Genes Virales/genética , Enfermedades de los Porcinos/virología , Virosis/veterinaria , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Clonación Molecular , Escherichia coli/genética , Datos de Secuencia Molecular , Homología de Secuencia de Aminoácido , Homología de Secuencia de Ácido Nucleico , Porcinos , Síndrome , Virosis/virología
18.
J Vet Diagn Invest ; 1(2): 101-4, 1989 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-2562190

RESUMEN

Stillborn and mummified swine fetuses from swine farms experiencing reproductive problems were investigated for evidence of infection with encephalomyocarditis (EMC) virus by fetal serology, virus isolation, and histopathologic examination. Fetal sera or thoracic fluids of 478 abnormal fetuses collected during January through December 1987 were tested for the presence of antibody specific to EMC virus. Of 478 samples tested, 175 (36.6%) had EMC virus serum neutralizing antibody titers of 1:64 or greater. The samples positive for EMC virus antibody were obtained from 38 swine farms located in 9 states in the United States. In addition to serologic observations, tissue samples of some abnormal fetuses were examined for the presence of virus and histopathologic lesions. The EMC virus was isolated in 1 case from the fetuses of an aborted litter. The isolate was serologically identical to a reference EMC virus. Nonsuppurative encephalitis and myocarditis were observed in the fetal samples collected from 2 different herds. Thoracic fluid of 1 stillborn pig with lesions was positive for EMC virus antibody at a titer of 1:512. We suggest that a widespread reproductive problem recently experienced in several major swine-producing areas of the United States may have been caused by EMC virus infection.


Asunto(s)
Virus de la Encefalomiocarditis/aislamiento & purificación , Infecciones por Enterovirus/veterinaria , Muerte Fetal/veterinaria , Enfermedades de los Porcinos/microbiología , Animales , Anticuerpos Antivirales/análisis , Anticuerpos Antivirales/sangre , Encéfalo/patología , Cerebelo/patología , Virus de la Encefalomiocarditis/inmunología , Infecciones por Enterovirus/inmunología , Infecciones por Enterovirus/microbiología , Infecciones por Enterovirus/patología , Muerte Fetal/microbiología , Técnica del Anticuerpo Fluorescente , Meninges/patología , Miocardio/patología , Porcinos , Enfermedades de los Porcinos/inmunología , Enfermedades de los Porcinos/patología , Tálamo/patología
19.
J Vet Diagn Invest ; 6(3): 289-92, 1994 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-7948196

RESUMEN

Various conditions were evaluated and modified to improve the sensitivity of the serum neutralization (SN) test for detecting antibody in pigs infected with porcine reproductive and respiratory syndrome virus (PRRSV). Higher SN titers were consistently obtained by the addition of 20% fresh swine serum to the virus diluent and by the use of a permissive cell clone (MARC-145) derived from the MA-104 cell line. Test sera used to assess the SN test were obtained from 2 groups of 3-week-old pigs infected intranasally with PRRSV (MN-1b). Using the modified method, SN antibody was first detected 9-11 days postinoculation (PI), with a peak evident at 11-21 days PI. The antibody subsequently declined, and a second peak was observed between 41 and 45 days PI. The first antibody peak was not observed and the SN antibody was only detectable between 32 and 41 days PI when the test was done with 20% heated swine serum or without supplemental swine serum. The SN antibody during 2-3 weeks PI was found to be sensitive to 2-mercaptoethanol or anti-swine IgM treatment. The SN antibody titers were high when homologous PRRSV isolate was used in the test but were markedly low for heterologous PRRSV isolates. No difference in antibody titers was observed when homologous and heterologous PRRSV isolates were tested by indirect fluorescent antibody assay. These results indicate that the modified SN method is useful in detecting earlier and higher PRRSV antibody and that it can differentiate among PRRSV isolates.


Asunto(s)
Anticuerpos Antivirales/aislamiento & purificación , Pruebas de Neutralización/veterinaria , Enfermedades de los Porcinos/inmunología , Virosis/veterinaria , Animales , Anticuerpos Antivirales/sangre , Técnica del Anticuerpo Fluorescente/veterinaria , Pruebas de Neutralización/métodos , Porcinos , Enfermedades de los Porcinos/sangre , Síndrome , Factores de Tiempo , Virosis/sangre , Virosis/inmunología
20.
J Vet Diagn Invest ; 5(2): 163-5, 1993 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-8507694

RESUMEN

The American and European strains of porcine reproductive and respiratory syndrome (PRRS) virus were initially isolated in an established cell line (CL 2621) and porcine alveolar macrophages (PAM), respectively. Subsequent isolation of American strains of this virus in PAM has also been reported. To determine their relative sensitivity for virus isolation, both PAM and CL 2621 cells were inoculated with 98 tissue specimens and 73 serum samples from animals suspected of having PRRS. Four of the 98 tissue samples yielded virus in both cell types, whereas 7 samples were positive only in PAM and 4 samples only in CL 2621. Of the 73 serum samples tested, 18 were positive in PAM of which only 2 were positive in CL 2621. Additionally, 82 isolates obtained initially in CL 2621 were inoculated in PAM cells, and 18 strains isolated originally in PAM were inoculated in CL 2621. Of the 82 CL 2621 isolates, 25 could not be propagated on PAM. Of the 57 that replicated in PAM, as detected by a positive test on indirect fluorescent antibody test, only 28 produced cytopathic effects and 29 did not. Of the 18 PAM isolates, 5 did not grow on CL 2621. Although PAM were relatively more sensitive for virus isolation, their failure to support the growth of certain strains of PRRS virus indicates the existence of variants among PRRS virus strains, and both PAM and CL 2621 should be used for virus isolation from clinical samples. In addition, the sensitivity of these 2 cell types was compared for the detection of fluorescent antibodies to PRRS virus using 179 serum samples from PRRS-infected animals.(ABSTRACT TRUNCATED AT 250 WORDS)


Asunto(s)
Anticuerpos Antivirales/análisis , Línea Celular/microbiología , Macrófagos Alveolares/microbiología , Enfermedades de los Porcinos/microbiología , Virus/aislamiento & purificación , Animales , Anticuerpos Antivirales/sangre , Infertilidad/microbiología , Infertilidad/veterinaria , Enfermedades Respiratorias/microbiología , Enfermedades Respiratorias/veterinaria , Sensibilidad y Especificidad , Porcinos , Síndrome
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