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1.
Parasitol Res ; 119(8): 2439-2462, 2020 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-32591866

RESUMEN

In 2018 and 2019, salmonid fishes, Salmo trutta L. and Salvelinus alpinus (L.) from lakes in Iceland were assessed for trematodes during a parasitological examination. Combined morphological and molecular analyses revealed the presence of four trematode species, two of which were previously known to parasitise salmonids in Iceland, Crepidostomum farionis (Müller, 1780) and Phyllodistomum umblae (Fabricius, 1780), and the two remaining species were recognised as new to science. Morphologically and genetically, Crepidostomum brinkmanni n. sp. and C. pseudofarionis n. sp. are closely related to two European species, namely C. metoecus and C. farionis. However, C. brinkmanni n. sp. is distinct by the position of maximum body width and arrangement of vitelline follicles; C. pseudofarionis n. sp. is distinct by its stout body, position of maximum body width, size of muscular papillae relative to oral sucker and the anterior extent of vitelline follicles. The new species were previously molecularly detected in their intermediate and definitive hosts in Norway and Ukraine, but their sequences were not supplemented with any morphological characterisation. In the present study, we provide detailed morphological descriptions and molecular sequences (28S rDNA and ITS2) of the four species of trematodes detected in Iceland. The discovery of the two new species of Crepidostomum indicates that the trematode diversity in fishes in the north is higher than previously known; our finding doubles the species spectrum of fish trematodes for Iceland. The record of C. brinkmanni from Ukraine indicates that its distribution might not be limited to northern latitudes.


Asunto(s)
Biodiversidad , Enfermedades de los Peces/parasitología , Salmonidae/parasitología , Trematodos/clasificación , Infecciones por Trematodos/parasitología , Animales , ADN Ribosómico/genética , Enfermedades de los Peces/epidemiología , Islandia/epidemiología , Lagos , Filogenia , Especificidad de la Especie , Trematodos/genética , Infecciones por Trematodos/epidemiología
3.
JEADV Clin Pract ; 3(1): 150-159, 2024 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-38646149

RESUMEN

Background: Beta-defensins (BDs) are antimicrobial peptides secreted upon epithelial injury. Both chemotactic and antimicrobial properties of BDs function as initial steps in host defense and prime the adaptive immune system in the body. Psoriasis, a chronic immune-mediated inflammatory disease, has both visible cutaneous manifestations as well as known associations with higher incidence of cardiometabolic complications and vascular inflammation. Objectives: We aimed to investigate the circulating expression of beta-defensin-2 (BD2) in psoriasis at baseline compared to control subjects, along with changes in BD2 levels following biologic treatment at one-year. The contribution of BD2 to subclinical atherosclerosis is also assessed. In addition, we have sought to unravel signaling mechanisms linking inflammation with BD2 expression. Methods: Multimodality imaging as well inflammatory biomarker assays were performed in biologic naïve psoriasis (n=71) and non-psoriasis (n=53) subjects. A subset of psoriasis patients were followed for one-year after biological intervention (anti-Tumor Necrosis Factor-α (TNFα), n=30; anti-Interleukin17A (IL17A), n=21). Measurements of circulating BD2 were completed by Enzyme-Linked Immunosorbent Assay (ELISA). Using HaCaT transformed keratinocytes, expression of BD2 upon cytokine treatment was assessed by quantitative polymerase chain reaction (qPCR) and ELISA. Results: Herein, we confirm that human circulating BD2 levels associate with psoriasis, which attenuate upon biologic interventions (anti-TNFα, anti-IL-17A). A link between circulating BD2 and sub-clinical atherosclerosis markers was not observed. Furthermore, we demonstrate that IL-17A-driven BD2 expression occurs in a Phosphatidylinositol 3-kinase (PI3-kinase) and Rac1 GTPase-dependent manner. Conclusions: Our findings expand on the potential role of BD2 as a tractable biomarker in psoriasis patients and describes the role of an IL-17A-PI3-kinase/Rac signaling axis in regulating BD2 levels in keratinocytes.

4.
J Clin Microbiol ; 50(3): 546-52, 2012 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-22219306

RESUMEN

Syphilis diagnosis is based on clinical observation, serological analysis, and dark-field microscopy (DFM) detection of Treponema pallidum subsp. pallidum, the etiological agent of syphilis, in skin ulcers. We performed a nested PCR (nPCR) assay specifically amplifying the tpp47 gene of T. pallidum from swab and blood specimens. We studied a cohort of 294 patients with suspected syphilis and 35 healthy volunteers. Eighty-seven of the 294 patients had primary syphilis, 103 had secondary syphilis, 40 had latent syphilis, and 64 were found not to have syphilis. The T. pallidum nPCR results for swab specimens were highly concordant with syphilis diagnosis, with a sensitivity of 82% and a specificity of 95%. Reasonable agreement was observed between the results obtained with the nPCR and DFM methods (kappa = 0.53). No agreement was found between the nPCR detection of T. pallidum in blood and the diagnosis of syphilis, with sensitivities of 29, 18, 14.7, and 24% and specificities of 96, 92, 93, and 97% for peripheral blood mononuclear cell (PBMC), plasma, serum, and whole-blood fractions, respectively. HIV status did not affect the frequency of T. pallidum detection in any of the specimens tested. Swab specimens from mucosal or skin lesions seemed to be more useful than blood for the efficient detection of the T. pallidum genome and, thus, for the diagnosis of syphilis.


Asunto(s)
Técnicas Bacteriológicas/métodos , Técnicas de Laboratorio Clínico/métodos , Técnicas de Diagnóstico Molecular/métodos , Reacción en Cadena de la Polimerasa/métodos , Sífilis/diagnóstico , Treponema pallidum/aislamiento & purificación , Adulto , Sangre/microbiología , Proteínas Portadoras/genética , Estudios de Cohortes , Femenino , Humanos , Lipoproteínas/genética , Masculino , Persona de Mediana Edad , Estudios Prospectivos , Sensibilidad y Especificidad , Úlcera Cutánea/microbiología , Treponema pallidum/genética
5.
J Invertebr Pathol ; 110(2): 174-83, 2012 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-22434000

RESUMEN

Shrimp farming in the Americas began to develop in the late 1970s into a significant industry. In its first decade of development, the technology used was simple and postlarvae (PLs) produced from wild adults and wild caught PLs were used for stocking farms. Prior to 1990, there were no World Animal Health Organization (OIE) listed diseases, but that changed rapidly commensurate with the phenomenal growth of the global shrimp farming industry. There was relatively little international trade of live or frozen commodity shrimp between Asia and the Americas in those early years, and with a few exceptions, most of the diseases known before 1980 were due to disease agents that were opportunistic or part of the shrimps' local environment. Tetrahedral baculovirosis, caused by Baculovirus penaei (BP), and necrotizing hepatopancreatitis (NHP) and its bacterial agent Hepatobacterium penaei, were among the "American" diseases that eventually became OIE listed and have not become established outside of the Americas. As the industry grew after 1980, a number of new diseases that soon became OIE listed, emerged in the Americas or were introduced from Asia. Spherical baculovirus, caused by MBV, although discovered in the Americas in imported live Penaeus monodon, was subsequently found to be common in wild and farmed Asian, Australian and African penaeids. Infectious hypodermal and hematopoietic necrosis virus (IHHNV) was introduced from the Philippines in the mid 1970s with live P. monodon and was eventually found throughout the Americas and subsequently in much of the shrimp farming industry in the eastern hemisphere. Taura syndrome emerged in Penaeus vannamei farms in 1991-1992 in Ecuador and was transferred to SE Asia with live shrimp by 1999 where it also caused severe losses. White Spot Disease (WSD) caused by White spot syndrome virus (WSSV) emerged in East Asia in ∼1992, and spread throughout most of the Asian shrimp farming industry by 1994. By 1995, WSSV reached the eastern USA via frozen commodity products and it reached the main shrimp farming countries of the Americas located on the Pacific side of the continents by the same mechanism in 1999. As is the case in Asia, WSD is the dominant disease problem of farmed shrimp in the Americas. The most recent disease to emerge in the Americas was infectious myonecrosis caused by IMN virus. As had happened before, within 3years of its discovery, the disease had been transferred to SE Asia with live P. vannamei, and because of its impact on the industry and potential for further spread in was listed by the OIE in 2005. Despite the huge negative impact of disease on the shrimp farming industry in the Americas, the industry has continued to grow and mature into a more sustainable industry. In marked contrast to 15-20years ago when PLs produced from wild adults and wild PLs were used to stock farms in the Americas, the industry now relies on domesticated lines of broodstock that have undergone selection for desirable characteristics including disease resistance.


Asunto(s)
Acuicultura/tendencias , Crustáceos/microbiología , Américas , Animales , Acuicultura/normas
6.
J Nutr Health Aging ; 26(11): 987-993, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-36437766

RESUMEN

OBJECTIVES: Older adults with poor oral health may be at higher risk of being pre-frail or frail. However, very few studies have examined this association in Latin American countries and middle-aged individuals. Therefore, we aimed to investigate the association between oral health and frailty status among Chilean adults ≥40 years. DESIGN: Cross-sectional study. SETTING AND PARTICIPANTS: We included 3,036 participants ≥40 years from the Chilean National Health Survey 2016-2017. METHODS: Frailty status was assessed with a 49-item frailty index, while the number of teeth, self-reported oral health, tooth decay, use of prostheses, and oral pain were the oral health conditions included. To assess the association between oral health conditions and frailty, we used multinomial logistic regression models status adjusted for sociodemographic and lifestyle variables. RESULTS: Overall, 40.6% and 11.8% of individuals were classified as pre-frail and frail, respectively. After adjusting for confounders, individuals with ≤20 teeth had a higher likelihood of being frail (odds ratio (OR): 1.94 [95% CI: 1.18-3.20]) than people with >20 teeth. Moreover, people with bad or very bad oral health, as well as oral pain, had a higher likelihood of being pre-frail (OR: 2.04 [95% CI: 1.40-2.97] and OR: 2.92 [95% CI: 1.58-5.39], respectively). Middle-aged individuals with fewer teeth and poor self-reported oral health had a higher likelihood of being pre-frail and frail than people ≥60. CONCLUSIONS AND IMPLICATIONS: Individuals with poor global oral health were more likely to be pre-frail or frail. This association seems to be stronger in people <60 years old. Our results are consistent with previously published reports.


Asunto(s)
Fragilidad , Humanos , Persona de Mediana Edad , Anciano , Fragilidad/epidemiología , Estudios Transversales , Salud Bucal , Oportunidad Relativa , Dolor
7.
Rev Med Chil ; 139(11): 1403-13, 2011 Nov.
Artículo en Español | MEDLINE | ID: mdl-22446644

RESUMEN

BACKGROUND: Clinical practice guidelines (CPG) are widely used as tools for improving quality of health care. Guidelines developed elsewhere, can be adapted using a valid and systematic process. AIM: To describe the methodology used in the process of adaptation of a guideline for the management of adults with community-acquired pneumonia (CAP) in a private health care organization. MATERIAL AND METHODS: We used the ADAPTE framework involving three main phases. At the set-up phase a guideline adaptation group integrated by medical specialists from different disciplines, a methodologist and a nurse coordinator was formed. At the adaptation phase, the specific clinical questions to be addressed by the guidelines were identified. RESULTS: Twenty five guidelines were initially retrieved. After their assessment, the number was reduced to only three. Recommendations from these guidelines were 'mapped' and focused searches were carried out where 'evidence gaps' were identified. An initial draft was written and revised by the adaptation group. At the finalization phase, the external review of the guideline was carried out and a process for the regular review and update of the adapted guideline was defined. CONCLUSIONS: We developed a guideline for the management of adults with CAP, adapted to the local context of our health care system, using guidelines developed elsewhere. This guideline creation method can be an efficient means of saving professional resources.


Asunto(s)
Comparación Transcultural , Atención a la Salud/organización & administración , Neumonía/terapia , Sector Privado/organización & administración , Sector Público/organización & administración , Adulto , Chile , Infecciones Comunitarias Adquiridas/terapia , Humanos
8.
Dev Biol (Basel) ; 126: 117-22; discussion 325-6, 2006.
Artículo en Inglés | MEDLINE | ID: mdl-17058487

RESUMEN

The most important diseases of farmed penaeid shrimp have infectious aetiologies. Among these are diseases with viral, rickettsial, bacterial, fungal and parasitic aetiologies. Diagnostic methods for these pathogens include the traditional methods of gross pathology, histopathology, classical microbiology, animal bioassay, antibody-based methods, and molecular methods using DNA probes and DNA amplification. While methods using clinical chemistry and tissue culture are standard methods in veterinary and human diagnostic laboratories, the former has not been routinely applied to the diagnosis of penaeid shrimp diseases and the latter has yet to be developed, despite considerable research and development efforts that have spanned the past 40 years. No continuous shrimp cell lines, or lines from other crustaceans, have been developed. Hence, when molecular methods began to be routinely applied to the diagnosis of infectious diseases in humans and domestic animals in the mid- to late 1980s, the technology was applied to the diagnosis of certain important diseases of penaeid shrimp for which only classical diagnostic methods were previously available. A DNA hybridization assay for the parvovirus IHHNV was the first molecular test developed for a shrimp disease. This was followed within a year by the first PCR test for MBV, an important baculovirus disease of shrimp. Today, shrimp disease diagnostic laboratories routinely use molecular tests for diagnostic and surveillance purposes for most of the important penaeid shrimp diseases.


Asunto(s)
Enfermedades de los Animales/diagnóstico , Enfermedades de los Animales/virología , Acuicultura/métodos , Técnicas de Diagnóstico Molecular/veterinaria , Penaeidae/virología , Enfermedades de los Animales/prevención & control , Animales , Técnicas de Diagnóstico Molecular/métodos , Virus/aislamiento & purificación
9.
Oncogene ; 18(35): 4974-82, 1999 Sep 02.
Artículo en Inglés | MEDLINE | ID: mdl-10490832

RESUMEN

The cell-cycle inhibitor p21 is upregulated during senescence and upon induction of senescence-like arrest by oncogenic Ras. We have used primary fibroblasts derived from p21-null mice to evaluate the role of p21 in these processes. We find that primary p21-/- cells enter senescence and have a lifespan similar to wild-type cells. Upon immortalization, most wild-type and p21-/- cultures acquire alterations in either p53 or p16INK4a, further indicating that p21-deficiency is not sufficient by itself to allow immortalization. Primary p21-/- cells, like wild-type cells, respond to oncogenic Ras by accumulating p53 and p16INK4a, and by decreasing their proliferation rate. In agreement with this, p21-/- cells are refractory to neoplasic transformation by oncogenic Ras when compared to p53-/- cells. We conclude that, in murine fibroblasts, p21 is not essential neither for senescence nor for preventing neoplasic transformation by oncogenic Ras.


Asunto(s)
Transformación Celular Neoplásica , Senescencia Celular , Ciclinas/fisiología , Fibroblastos/citología , Proteína Oncogénica p21(ras)/metabolismo , Células 3T3 , Animales , Ciclo Celular , División Celular , Línea Celular Transformada , Transformación Celular Neoplásica/genética , Células Cultivadas , Inhibidor p16 de la Quinasa Dependiente de Ciclina/genética , Inhibidor p16 de la Quinasa Dependiente de Ciclina/metabolismo , Inhibidor p21 de las Quinasas Dependientes de la Ciclina , Ciclinas/deficiencia , Ciclinas/genética , Doxorrubicina/farmacología , Fibroblastos/efectos de los fármacos , Fibroblastos/metabolismo , Eliminación de Gen , Expresión Génica/efectos de los fármacos , Ratones , Ratones Noqueados , Proteína Oncogénica p21(ras)/genética , Pase Seriado , Transducción Genética , Proteína p53 Supresora de Tumor/genética , Proteína p53 Supresora de Tumor/metabolismo
10.
J Parasitol ; 101(4): 492-9, 2015 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-25885554

RESUMEN

Morphological and genetic data on the larvae of Anisakidae parasitizing the sandperches Pseudopercis numida and Pinguipes brasilianus were recorded from Cabo Frio, State of Rio de Janeiro, Brazil (23°S, 42°W). Twenty-five specimens of P. numida and 25 specimens of P. brasilianus were necropsied. The 352 larvae collected included 1 Anisakis sp., 344 Hysterothylacium sp., and 7 Raphidascaris sp. We present mtDNA sequence data for Anisakis sp. and Hysterothylacium sp., and we sequenced the ITS ribosomal DNA (rDNA) of Hysterothylacium sp. and Raphidascaris sp. The morphological and genetic profiles confirmed the identification of Anisakis typica (Diesing, 1860). Hysterothylacium sp. and Raphidascaris sp. were not identified to the species level due to the absence of similar sequences for adult parasites. The mtDNA sequence of Hysterothylacium sp. from P. numida showed a high similarity to the sequence of Hysterothylacium deardorffoverstreetorum. However, a phylogenetic analysis demonstrated the presence of different clades under the same name as the larval H. deardorffoverstreetorum. Pseudopercis numida is a new host record for A. typica.


Asunto(s)
Infecciones por Ascaridida/veterinaria , Ascaridoidea/clasificación , Enfermedades de los Peces/parasitología , Percas/parasitología , Animales , Infecciones por Ascaridida/parasitología , Ascaridoidea/anatomía & histología , Ascaridoidea/genética , Secuencia de Bases , Brasil , ADN de Helmintos/química , ADN Mitocondrial/química , ADN Ribosómico/química , Funciones de Verosimilitud , Datos de Secuencia Molecular
11.
Exp Gerontol ; 36(8): 1289-302, 2001 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-11602205

RESUMEN

The tumor suppressor gene ARF is formed by three exons, namely exons 1 beta, 2 and 3. Here, we show that embryo fibroblasts from mice genetically deficient in exons 2 and 3 (Delta 2,3) express a transcript formed by exon 1 beta followed by the 3'-terminal exon of the gene immediately downstream of the INK4a/ARF locus, which we have called NTp16 (Next-To-p16). The chimeric ARF-NTp16 transcript is not detectable in wild-type fibroblasts but its expression level in Delta 2,3 fibroblasts is 30% compared to the level of the normal ARF transcript in wild-type cells. Expression of the ARF-NTp16 transcript in Delta 2,3 cells is subject to normal regulatory features, such as upregulation by the accumulation of cell doublings, and by the presence of oncogenic Ras or E1a. The chimeric ARF-NTp16 transcript has the potential to encode a 17kDa peptide; however, this peptide is not accumulated in cells at detectable levels, probably reflecting poor codon usage or protein instability. We conclude that Delta 2,3 cells do not retain ARF functionality, at least to a significant extent. Interestingly, the expression pattern of the full-length NTp16 gene is altered in several tissues by the presence of the Delta 2,3 mutation. Finally, these data identify the gene immediately downstream of the INK4a/ARF locus, a region that has been previously proposed to contain another tumor suppressor different from the INK4a/ARF genes.


Asunto(s)
Inhibidor p16 de la Quinasa Dependiente de Ciclina/genética , Genes Supresores de Tumor , Genes p16 , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Células Cultivadas , Quimera/genética , Mapeo Cromosómico , Cartilla de ADN/genética , ADN Complementario/genética , Exones , Expresión Génica , Ratones , Ratones Noqueados , Datos de Secuencia Molecular , Mutación
12.
Ann N Y Acad Sci ; 916: 628-34, 2000.
Artículo en Inglés | MEDLINE | ID: mdl-11193688

RESUMEN

We studied the levels of resistance to seven insecticides: malathion, chlorpyrifos, pirimiphos-methyl, propoxur, cypermethrin, deltamethrin, and lambda-cyhalothrin in nine strains of Blattella germanica (Linnaeus, 1717) collected from sites in Santiago de Cuba and Havana City. The strains from Santiago de Cuba, generally had high levels of resistance to malathion, cypermethrin, deltamethrin, and lambda-cyhalothrin, but only low levels of resistance to pirimiphos-methyl, chlorpyrifos, and propoxur. In the strains from Havana City we found a moderate resistance to the organophosphate insecticides, resistance to the pyrethroids (except for three strains that showed susceptibility to lambda-cyhalothrin), and susceptibility to the carbamate insecticide (propoxur).


Asunto(s)
Blattellidae , Resistencia a los Insecticidas , Insecticidas/toxicidad , Animales , Cloropirifos/toxicidad , Cuba , Geografía , Malatión/toxicidad , Nitrilos , Compuestos Organotiofosforados/toxicidad , Propoxur/toxicidad , Piretrinas/toxicidad , Especificidad de la Especie
13.
Oncol Rep ; 8(2): 425-9, 2001.
Artículo en Inglés | MEDLINE | ID: mdl-11182068

RESUMEN

IFNs are a family of cytokines involved in antiviral defense, cell growth regulation and immune activation. IFNs either inhibit cell proliferation or control apoptosis depending on factors such as cell type and state of cell differentiation. It is important to determine how IFN-induced gene products interact with other cellular proteins to produce these responses. We have investigated the effect of IFNalpha 2b on a human small cell lung carcinoma (SCLC) cell line H82. We have found that IFNalpha efficiently induces apoptosis in H82 cells. The induction of apoptosis by IFNalpha 2b is accompanied by decreased levels of c-myc and Cdk2. We have also observed that in H82 cells IFNalpha induces downregulation of p27 and this is in contrast to the upregulation of p27 observed in other cell types where IFNs induce cell cycle arrest. IFNalpha-induced downregulation of p27 is due to protein destabilization and can be prevented by the proteasome inhibitor LLnL. The data suggest that in H82 cells, IFNalpha 2b induces degradation of p27Kip1 independently of CDK2 kinase activity and through a ubiquitin or ubiquitin-related pathway and that the degradation of p27Kip1 could be a molecular event of importance for IFN-induced apoptosis in cancer cells.


Asunto(s)
Apoptosis/efectos de los fármacos , Quinasas CDC2-CDC28 , Proteínas de Ciclo Celular , Cisteína Endopeptidasas/metabolismo , Interferón-alfa/farmacología , Proteínas Asociadas a Microtúbulos/metabolismo , Complejos Multienzimáticos/metabolismo , Proteínas Supresoras de Tumor , Carcinoma de Células Pequeñas , Quinasa 2 Dependiente de la Ciclina , Inhibidor p27 de las Quinasas Dependientes de la Ciclina , Quinasas Ciclina-Dependientes/metabolismo , Inhibidores Enzimáticos/metabolismo , Genes myc , Humanos , Interferón alfa-2 , Neoplasias Pulmonares , Inhibidores de Proteasas/farmacología , Complejo de la Endopetidasa Proteasomal , Proteínas Serina-Treonina Quinasas/metabolismo , Proteínas Proto-Oncogénicas c-myc/metabolismo , Proteínas Recombinantes , Células Tumorales Cultivadas
14.
Dis Aquat Organ ; 44(2): 87-96, 2001 Mar 09.
Artículo en Inglés | MEDLINE | ID: mdl-11324820

RESUMEN

A post-embedding in situ hybridization procedure was developed to detect hepatopancreatic parvovirus (HPV) of penaeid shrimp at the ultrastructural level. The procedure was optimized using sections of resin-embedded hepatopancreas from HPV-infected juvenile Penaeus monodon and postlarval P. chinensis. The hepatopancreata were fixed using various fixatives, dehydrated, and embedded in the hydrophilic resin Unicryl. A 592 bp HPV-specific DNA probe, labeled with DIG-11-dUTP, was tested both on semi-thin and ultra-thin sections and examined by light and electron microscopy, respectively. Hybridized probe was detected by means of an anti-DIG antibody conjugated to 10 nm gold particles and subsequent silver enhancement. Hybridization signal intensities were similar with all fixatives tested, but ultrastructure was best preserved with either 2 or 6% glutaraldehyde. Post-fixation with 1% osmium tetroxide improved ultrastructure but markedly decreased hybridization signal and induced non-specific deposition of gold and silver. Under optimized conditions, this technique was used to successfully follow the development of HPV from absorption and transport through the cytoplansm to nuclear penetration, replication and release by cytolysis. The probe signal was consistently observed among necrotic cell debris within the lumen of hepatopancreatic tubules, within the microvillous border of tubule epithelial cells, within the cytoplasm, and within diagnostic HPV intranuclear inclusion bodies. The nucleolus and karyoplasm of patently infected cells (i.e., showing HPV intranuclear inclusion bodies) were almost devoid of signal. Electron-lucent structures, known as intranuclear bodies, commonly found within the virogenic stroma, showed only weak labeling. This is the first use of in situ hybridization to detect HPV nucleic acids with the electron microscope. The technique should be useful for studying the pathogenesis of HPV.


Asunto(s)
Decápodos/virología , Densovirinae/aislamiento & purificación , Hibridación in Situ/veterinaria , Animales , Decápodos/ultraestructura , Microscopía Electrónica/veterinaria
15.
Dis Aquat Organ ; 39(3): 177-82, 2000 Feb 09.
Artículo en Inglés | MEDLINE | ID: mdl-10768285

RESUMEN

Current methods to detect hepatopancreatic parvovirus (HPV) infection of penaeid shrimp depend on invasive techniques that require dissecting the organs infected by this virus. However, sacrificing valuable stocks in order to determine their HPV status can be a drawback in the case of breeding programs. A method was developed for HPV detection by applying a polymerase chain reaction (PCR) assay to fecal samples collected from live HPV-infected shrimp Penaeus chinensis. A pair of PCR primers, 1120F/1120R, which amplify a 592 base pair (bp) region from the virus genome, was designed from previously known HPV sequence information (HPV clone HPV8). PCR amplification with these primers generated a product of the expected size directly from the crude feces of HPV-infected shrimp but not from the feces of specific pathogen-free (SPF) shrimp. The HPV origin of the amplified product was validated by means of an in situ hybridization assay where the product of the amplification, labeled with digoxigenin (DIG)-11-dUTP, showed an intense reaction within hepatopancreatic cells displaying characteristic HPV lesions on HPV-infected shrimp. No reaction to this probe was observed when reacted in situ with sections of the hepatopancreas of SPF specimens or to sections of shrimp infected by the infectious hypodermal and hematopoietic necrosis virus (IHHNV), another parvovirus of penaeid shrimp. These primers were tested for specificity against homologous and nonhomologous viruses and no product was amplified. A fragment of the expected size was obtained only when purified HPV or purified HPV8 plasmid was used as template DNA. Under optimized conditions, these primers detected as little as 1 fg of purified HPV8 plasmid DNA, equivalent to approximately 300 HPV particles. Analysis of fecal samples by PCR may prove useful for non-lethal screening of valuable shrimp of unknown HPV status. This same strategy also might be used for detection of other enteric viruses that infect penaeid shrimp.


Asunto(s)
Decápodos/virología , Parvovirus/aislamiento & purificación , Animales , Cartilla de ADN/química , Sondas de ADN/química , ADN Viral/química , Sistema Digestivo/patología , Sistema Digestivo/virología , Electroforesis en Gel de Agar , Heces/virología , Histocitoquímica , Hibridación in Situ/veterinaria , Cuerpos de Inclusión Viral/patología , Parvovirus/química , Parvovirus/genética , Reacción en Cadena de la Polimerasa/métodos , Reacción en Cadena de la Polimerasa/veterinaria , Sensibilidad y Especificidad , Organismos Libres de Patógenos Específicos
16.
Dis Aquat Organ ; 47(1): 13-23, 2001 Oct 29.
Artículo en Inglés | MEDLINE | ID: mdl-11797911

RESUMEN

Monoclonal antibodies (MAbs) were produced against white spot syndrome virus (WSSV) of penaeid shrimp. The virus isolate used for immunization was obtained from China in 1994 and was passaged in Penaeus vannamei. The 4 hybridomas selected for characterization all produced MAbs that reacted with the 28 kD structural protein by Western blot analysis. The MAbs tested in dot-immunoblot assays were capable of detecting the virus in hemolymph samples collected from moribund shrimp during an experimentally induced WSSV infection. Two of the MAbs were chosen for development of serological detection methods for WSSV. The 2 MAbs detected WSSV infections in fresh tissue impression smears using a fluorescent antibody for final detection. A rapid immunohistochemical method using the MAbs on Davidson's fixed tissue sections identified WSSV-infected cells and tissues in a pattern similar to that seen with digoxigenin-labeled WSSV-specific gene probes. A whole mount assay of pieces of fixed tissue without paraffin embedding and sectioning was also successfully used for detecting the virus. None of the MAbs reacted with hemolymph from specific pathogen-free shrimp or from shrimp infected with infectious hypodermal and hematopoietic necrosis virus, yellow head virus or Taura syndrome virus. In Western blot analysis, the 2 MAbs did not detect any serological differences among WSSV isolates from China, Thailand, India, Texas, South Carolina or Panama. Additionally, the MAbs did not detect a serological difference between WSSV isolated from penaeid shrimp and WSSV isolated from freshwater crayfish.


Asunto(s)
Anticuerpos Monoclonales , Virus ADN/inmunología , Decápodos/virología , Animales , Anticuerpos Monoclonales/biosíntesis , Western Blotting/veterinaria , Virus ADN/aislamiento & purificación , Electroforesis en Gel de Poliacrilamida/veterinaria , Técnica del Anticuerpo Fluorescente/veterinaria , Hemolinfa , Immunoblotting/veterinaria , Inmunohistoquímica/veterinaria , Organismos Libres de Patógenos Específicos
17.
Dis Aquat Organ ; 46(2): 153-8, 2001 Sep 12.
Artículo en Inglés | MEDLINE | ID: mdl-11678231

RESUMEN

Hepatopancreatic parvovirus (HPV) can cause stunted growth and death in penaeid shrimp including Penaeus monodon. We used PCR primers and a commercial DNA probe designed from HPV of Penaeus chinensis (HPVchin) to examine HPV-infected Thai P. monodon (HPVmon). We found that the PCR primers produced a 732 bp DNA amplicon rather than the 350 bp amplicon obtained with HPVchin template and that the DNA probe gave weak to variable in situ DNA hybridization results. In addition, hybridization to PCR products from HPVmon was weak compared with hybridization with PCR products from HPVchin. By contrast, the 732 bp amplicon hybridized strongly with HPVmon-infected cells by in situ hybridization but not with uninfected shrimp tissue or other shrimp viruses, thus confirming its origin from HPVmon. Cloning, sequencing and analysis of the 732 bp amplicon showed that 696 bp (excluding the primer sequences) contained 47% GC content and had only 78% homology to 701 aligned bases from a 3350 bp DNA fragment of HPVchin from GenBank. These results explain why the reagents based on HPVchin gave a different PCR product and weak hybridization results with HPVmon, and they show that multiple primers or degenerate primers may be necessary for general detection of HPV varieties. Together with previously published information on the estimated total genome sizes for HPVchin (approximately 4 kb) and HPVmon (approximately 6 kb), these data support the contention that HPVchin and HPVmon are different varieties or species, in spite of their similar histopathology.


Asunto(s)
ADN Viral/análisis , Decápodos/virología , Parvovirus/clasificación , Parvovirus/aislamiento & purificación , Animales , Cartilla de ADN , Sondas de ADN , Electroforesis en Gel de Agar/veterinaria , Amplificación de Genes , Hibridación in Situ , Peso Molecular , Parvovirus/genética , Filogenia , Reacción en Cadena de la Polimerasa/veterinaria , Juego de Reactivos para Diagnóstico/veterinaria , Sensibilidad y Especificidad , Alineación de Secuencia , Homología de Secuencia de Ácido Nucleico
18.
J Ethnopharmacol ; 37(2): 145-9, 1992 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-1434688

RESUMEN

The objectives of this work were to investigate the effects of a garlic dialysate on diastolic blood pressure (DBP), heart rate (HR) and electrocardiogram (ECG) of anaesthetized dogs and its effects on frequency and tension of isolated rat atria. Garlic dialysate led to a drop in DBP (from 112.5 +/- 3.67 to 70 +/- 3.16 mmHg) and a decrease in HR (from 198 +/- 9.81 to 164 +/- 16.59 beats/min) in a dose-dependent manner. The ECG showed a regular sinus bradycardic rhythm. The addition of garlic dialysate to isolated left rat atria evoked a decrease in tension development. Frequency, measured by spontaneous beating of the right atria, was also reduced. Both effects were dose-dependent. In addition to these effects, the positive inotropism and chronotropism induced by addition of isoproterenol 10(-9) M, were partially antagonized by preincubation of the rat atria with the garlic dialysate. The above findings can be explained by a depressant effect on automaticity and tension development in the heart, suggesting a beta-adrenoceptor blocking action produced by the garlic dialysate.


Asunto(s)
Antiarrítmicos/farmacología , Ajo/química , Plantas Medicinales , Anestesia , Animales , Presión Sanguínea/efectos de los fármacos , Diálisis , Perros , Electrocardiografía/efectos de los fármacos , Femenino , Corazón/efectos de los fármacos , Frecuencia Cardíaca/efectos de los fármacos , Técnicas In Vitro , Isoproterenol/farmacología , Masculino , Músculo Liso Vascular/efectos de los fármacos , Contracción Miocárdica/efectos de los fármacos , Extractos Vegetales/farmacología , Ratas
19.
J Ethnopharmacol ; 52(2): 101-5, 1996 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-8735454

RESUMEN

The intravenous administration of chromatographically purified fractions of garlic (2, 4 and 6 micrograms/kg dry weight) to anaesthetized rabbits elicits dose-dependent diuretic-natriuretic responses which reach a maximum 60 min after injection, and return to basal levels after 90 min. A gradual decrease in heart rate, but not in arterial blood pressure was observed during the course of the experimental periods. The electrocardiogram was not affected.


Asunto(s)
Diuresis/efectos de los fármacos , Ajo/metabolismo , Natriuresis/efectos de los fármacos , Plantas Medicinales , Animales , Presión Sanguínea/efectos de los fármacos , Fraccionamiento Químico , Cromatografía en Gel , Relación Dosis-Respuesta a Droga , Electrocardiografía/efectos de los fármacos , Frecuencia Cardíaca/efectos de los fármacos , Inyecciones Intravenosas , Masculino , Extractos Vegetales/administración & dosificación , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/farmacología , Conejos
20.
J Ethnopharmacol ; 31(3): 325-31, 1991 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-2056760

RESUMEN

Gastric administration of encapsuled garlic powder to anaesthetized dogs induced dose-dependent (2.5 to 15 mg/kg) natriuretic and diuretic responses which reached maximum 30-40 min after garlic administration and decreased to basal levels after 100-150 min. A simultaneous decrease in arterial blood pressure was observed which continued past the 250 min-mark. High garlic doses (15 and 20 mg/kg) provoked bradycardia and T-wave inversion during the first 10-15 min of the experiment with recordings returning to normal and staying normal throughout the remainder of the experiment.


Asunto(s)
Antihipertensivos , Diuréticos , Ajo , Natriuresis/efectos de los fármacos , Plantas Medicinales , Anestesia , Animales , Presión Sanguínea/efectos de los fármacos , Perros , Electrocardiografía , Femenino , Frecuencia Cardíaca/efectos de los fármacos , Masculino
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