RESUMEN
A large outbreak of enterohaemorrhagic Escherichia coli (EHEC) O111 and O157 occurred in Japan in April 2011. We conducted an unmatched case-control study and trace-back investigation to determine the source of EHEC O111 infection and risk factors for severe complications. Pulsed-field gel electrophoresis was performed to help define cases. A total of 86 individuals met the case definition. Of these, 40% experienced haemolytic uraemic syndrome (HUS), 24% acute encephalopathy, and 6% died. Illness was significantly associated with eating the raw beef dish yukhoe (odds ratio 19·64, 95% confidence interval 7·03-54·83), the likely food vehicle. EHEC O111 and its closely related stx-negative variants were found in the beef. HUS occurred most frequently in individuals aged 5-9 years, and this age group was significantly associated with acute encephalopathy. The prevalence of HUS and acute encephalopathy was higher than in previous non-O157-related outbreaks, indicating a high risk of severe complications.
Asunto(s)
Encefalopatías/epidemiología , Encefalopatías/microbiología , Brotes de Enfermedades , Escherichia coli Enterohemorrágica/aislamiento & purificación , Infecciones por Escherichia coli/epidemiología , Infecciones por Escherichia coli/microbiología , Enfermedades Transmitidas por los Alimentos/epidemiología , Enfermedades Transmitidas por los Alimentos/microbiología , Síndrome Hemolítico-Urémico/epidemiología , Síndrome Hemolítico-Urémico/microbiología , Carne/microbiología , Enfermedad Aguda , Adolescente , Adulto , Estudios de Casos y Controles , Niño , Preescolar , Electroforesis en Gel de Campo Pulsado , Escherichia coli O157/aislamiento & purificación , Femenino , Microbiología de Alimentos , Humanos , Lactante , Japón/epidemiología , Masculino , Persona de Mediana Edad , Factores de RiesgoRESUMEN
Cardiac surgery with cardiopulmonary bypass is associated with the development of a systemic inflammatory response that can often lead to dysfunction of major organs. We hypothesised that the highly selective α2-adrenergic agonist, dexmedetomidine, attenuates the systemic inflammatory response. Forty-two patients were randomly assigned to receive dexmedetomidine or saline after aortic cross-clamping). The mean (SD) levels of the nuclear protein plasma high-mobility group box 1 increased significantly from 5.1 (2.2) ng ml(-1) during (16.6 (7.3) ng ml(-1) ) and after (14.3 (8.2) ng ml(-1) ) cardiopulmonary bypass in the saline group. In the dexmedetomidine group, the levels increased significantly only during cardiopulmonary bypass (4.0 (1.9) ng ml(-1) baseline vs. 10.8 (2.7) ng ml(-1) ) but not after (7.4 (3.8) ng ml(-1) ). Dexmedetomidine infusion also suppressed the rise in mean (SD) interleukin-6 levels after cardiopulmonary bypass (a rise of 124.5 (72.0) pg ml(-1) vs. 65.3 (30.9) pg ml(-1)). These suppressive effects of dexmedetomidine might be due to the inhibition of nuclear factor kappa B activation and suggest that intra-operative dexmedetomidine may beneficially inhibit inflammatory responses associated with ischaemia-reperfusion injury during cardiopulmonary bypass.
Asunto(s)
Analgésicos no Narcóticos/farmacología , Puente Cardiopulmonar/efectos adversos , Dexmedetomidina/farmacología , Mediadores de Inflamación/sangre , Cuidados Posoperatorios/métodos , Síndrome de Respuesta Inflamatoria Sistémica/sangre , Síndrome de Respuesta Inflamatoria Sistémica/prevención & control , Anciano , Analgésicos no Narcóticos/sangre , Biomarcadores/sangre , Dexmedetomidina/sangre , Femenino , Humanos , Interleucina-6/sangre , Masculino , FN-kappa B/sangre , FN-kappa B/efectos de los fármacos , Estudios Prospectivos , Cloruro de Sodio/administración & dosificación , Síndrome de Respuesta Inflamatoria Sistémica/etiología , Factores de Tiempo , Resultado del TratamientoRESUMEN
An abnormal lipoprotein was visualized directly in serum by electron microscopy of preparations negatively stained with potassium phosphotungstate. It appears as a unique disk-shaped particle with major axis measuring 400 to 600 angstroms and minor axis measuring about 100 angstroms. Chemical analysis, viscosity measurements, and x-ray diffraction analysis of purified preparations indicate that the particle, consisting of a one-to-one molar mixture of cholesterol and choline phosphatides associated with a small amount of protein, is a flattened vesicle, the wall of which is a continuous lipid bilayer.
Asunto(s)
Trastornos de las Proteínas Sanguíneas/sangre , Colestasis/sangre , Hiperlipidemias/sangre , Lipoproteínas/sangre , Colesterol/sangre , Ésteres/sangre , Humanos , Microscopía Electrónica , Fosfolípidos/sangre , Ácido Fosfotúngstico , Gravedad Específica , Triglicéridos/sangre , Ultracentrifugación , Viscosidad , Difracción de Rayos XRESUMEN
We investigated the level of telomerase activity (TA) in 17 specimens of non-genital Bowen's disease (BD) and in 14 specimens of skin without sun exposure (non-exposed skin) using a non-isotopic PCR-based telomeric repeat amplification protocol (TRAP) assay. Expression of human telomerase reverse transcriptase (hTERT; the catalytic subunit of telomerase) was also evaluated by immunochemistry in the non-genital BD tissues. Moderate to high levels of TA were detected in 41.2% of 17 non-genital BD specimens (P = 0.001). In contrast, TA was not evident in non-exposed skin. Recently, nucleolin was reported to be associated with hTERT, so we used this antibody instead of hTERT antibody. Immunohistochemistry showed that nucleolin expression was associated with high TA levels in non-genital BD. Our results also revealed differences of TA levels among non-genital BD specimens. High levels of TA in those specimens were not age related. Five out of 7 specimens (71.4%) with moderate to high TA levels were from sun-exposed sites, while the remaining 10 specimens with low levels of TA were from non-exposed sites. These results suggested that cellular DNA damage caused by ultraviolet irradiation might be associated with an increase of TA in non-genital BD. Among non-genital BD specimens, 4 out of 17 (23.5%) showed high levels of TA (median relative TA value: 79.8%; P = 0.003), which might be associated with immortalization or transformation to invasive squamous cell carcinoma.
Asunto(s)
Enfermedad de Bowen/enzimología , Telomerasa/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Secuencia de Bases , Cartilla de ADN , Humanos , Inmunohistoquímica , Persona de Mediana Edad , Reacción en Cadena de la PolimerasaRESUMEN
An ATM-dependent cellular signal, a DNA-damage response, has been shown to be involved during infection of human immunodeficiency virus type-1 (HIV-1), and a high incidence of malignant tumor development has been observed in HIV-1-positive patients. Vpr, an accessory gene product of HIV-1, delays the progression of the cell cycle at the G2/M phase, and ATR-Chk1-Wee-1, another DNA-damage signal, is a proposed cellular pathway responsible for the Vpr-induced cell cycle arrest. In this study, we present evidence that Vpr also activates ATM, and induces expression of gamma-H2AX and phosphorylation of Chk2. Strikingly, Vpr was found to stimulate the focus formation of Rad51 and BRCA1, which are involved in repair of DNA double-strand breaks (DSBs) by homologous recombination (HR), and biochemical analysis revealed that Vpr dissociates the interaction of p53 and Rad51 in the chromatin fraction, as observed under irradiation-induced DSBs. Vpr was consistently found to increase the rate of HR in the locus of I-SceI, a rare cutting-enzyme site that had been introduced into the genome. An increase of the HR rate enhanced by Vpr was attenuated by an ATM inhibitor, KU55933, suggesting that Vpr-induced DSBs activate ATM-dependent cellular signal that enhances the intracellular recombination potential. In context with a recent report that KU55933 attenuated the integration of HIV-1 into host genomes, we discuss the possible role of Vpr-induced DSBs in viral integration and also in HIV-1 associated malignancy.
Asunto(s)
Proteínas de Ciclo Celular/metabolismo , Proteínas de Unión al ADN/metabolismo , Productos del Gen vpr/fisiología , Proteínas Serina-Treonina Quinasas/metabolismo , Recombinación Genética , Proteínas Supresoras de Tumor/metabolismo , Proteínas de la Ataxia Telangiectasia Mutada , Células Cultivadas , ADN/metabolismo , Roturas del ADN de Doble Cadena , Reparación del ADN , Regulación de la Expresión Génica , Humanos , Transducción de SeñalRESUMEN
As an extension of metabolic studies of the cholesteryl ester component of rat very low density lipoproteins, we have studied the metabolism of the B apoprotein component labeled by intravenous injection of [3H]lysine. The B apoprotein separated from other apoproteins by delipidation and selective precipitation with tetramethylurea could not be distinguished from B apoprotein prepared by the conventional gel filtration technique. After injection of [3H]lysine, specific activity of B apoprotein was maximal in very low density and low density lipoproteins 1 and 11/2-h later, respectively, in a manner consistent with a precursor-product relationship. When protein-labeled very low density lipoproteins were injected into rats, the relationships of specific activity again indicated that B apoprotein of very low density lipoproteins may be the sole precursor of that of low density lipoproteins. However, less than 10% of the B apoprotein that disappeared from very low density lipoproteins appeared in density lipoproteins. To evaluate the sites of removal of B aproprotein of very low density lipoproteins from plasma, protein-labeled very low density lipoproteins were incubated with unlabeled high density lipoproteins to reduce radioactivity in non-B apoproteins selectively by molecular exchange. Most of the B apoprotein was rapidly removed by the liver. The extensive hepatic uptake of both the cholesteryl ester and B apoprotein components of rat very low density lipoproteins may explain the characteristically low concentrations of plasma low density lipoproteins in the rat.
Asunto(s)
Apoproteínas/metabolismo , Lipoproteínas VLDL/metabolismo , Hígado/metabolismo , Aminoácidos , Animales , Fenómenos Químicos , Química , Ésteres del Colesterol/metabolismo , Cromatografía en Gel , Electroforesis en Gel de Poliacrilamida , Lipoproteínas LDL/metabolismo , Lipoproteínas VLDL/aislamiento & purificación , Lisina/metabolismo , Masculino , RatasRESUMEN
Methods for quantitation of the major apoproteins of human serum very low density lipoprotein have been developed employing tetramethylurea, which delipidates the lipoprotein and selectively precipitates apolipoprotein B. Six soluble apoproteins are separated by electrophoresis in polyacrylamide gel. One of these is a previously unrecognized species of R-alanine (R4-alanine), more anionic than the R3-alanine polypeptide. Conditions of staining have been found which yield reproducibly linear chromogenic response with native lipoprotein and with each purified apoprotein. Recovery of protein in the seven species measured accounts for over 97% of the total in the very low density lipoprotein of normolipidemic individuals and in most samples from individuals with endogenous hyperlipemia. The mean content of apolipoprotein B in 43 samples from normolipidemic subjects was 36.9(+/-1.2 SEM)% of total protein, The distribution of the major soluble apoproteins as mean (+/-SEM) percentage of the soluble fraction was : R-serine, 5.3+/-o.5; arginine-rich, 20.6+/-1.0; R-glutamic, 10.6+/-0.4; R2-alanine, 28.3+/-0.7; R3-alanine, 26.9+/-0.5; and R4-alanine, 8.0+/-0.5. Distribution of the apoproteins was a function of particle diameter of very low density lipoprotein in fractions separated by gel permeation chromatography and by density gradient ultracentrifugation. In fractions below 700-800 A, apolipoprotein B comprised an increasing percentage of the total protein with decreasing particle diameter. Among the soluble proteins the percentage of the arginine-rich and R-serine polypeptides increased and that of the R-glutamic polypeptide declined progressively with decreasing particle size. Apoprotein distribution was similar in fractions of similar particle size from normolipidemic and hyperlipemic subjects with the exception that all fractions from the hyperlipemic subjects contained more R-serine and some, more arginine rich polypeptide. Even in the absence of chylomicrons, the distribution of soluble apoproteins in particles of diameters greater than 700-800 A was usually similar to that of the smallest particles. This suggests that the largest particles may include products of the partial catabolism of chylomicrons.
Asunto(s)
Apoproteínas/análisis , Hiperlipidemias/sangre , Lipoproteínas VLDL/análisis , Adulto , Centrifugación por Gradiente de Densidad , Fenómenos Químicos , Química , Cromatografía , Electroforesis en Gel de Poliacrilamida , Humanos , Lipoproteínas VLDL/sangre , Persona de Mediana Edad , UltracentrifugaciónRESUMEN
Monoclonal antibodies to the prion protein (PrP) have been of critical importance in the neuropathological characterization of PrP-related disease in men and animals. To determine the influence of species-specific amino-acid substitutions recognized by monoclonal antibodies, and to investigate the immunohistochemical reactivity of the latter, analyses were carried out on brain sections of cattle with bovine spongiform encephalopathy, sheep with scrapie, mice infected with scrapie, and human beings with Creutzfeldt-Jakob disease (CJD) or Gerstmann-Sträussler-Sheinker disease (GSS). Immunoreactivity varied between the antibodies, probably as the result of differences in the amino-acid sequence of the prion protein in the various species. Some monoclonal antibodies against mouse recombinant PrP gave strong signals with bovine, ovine and human PrP(Sc), in addition to murine PrP(Sc), even though the amino-acid sequences determined by the antibody epitope are not fully identical with the amino-acid sequences proper to the species. On the other hand, in certain regions of the PrP sequence, when the species-specificity of the antibodies is defined by one amino-acid substitution, the antibodies revealed no reactivity with other animal species. In the region corresponding to positions 134-159 of murine PrP, immunohistochemical reactivity or species-specificity recognized by the antibodies may be determined by one amino acid corresponding to position 144 of murine PrP. Not all epitopes recognized by a monoclonal antibody play an important role in antigen-antibody reactions in immunohistochemistry. The presence of the core epitope is therefore vital in understanding antibody binding ability.
Asunto(s)
Anticuerpos Monoclonales/inmunología , Especificidad de Anticuerpos , Inmunohistoquímica/métodos , Enfermedades por Prión/inmunología , Enfermedades por Prión/veterinaria , Priones/inmunología , Secuencia de Aminoácidos , Animales , Encéfalo/metabolismo , Encéfalo/patología , Bovinos , Epítopos/inmunología , Humanos , Ratones , Datos de Secuencia Molecular , Ovinos , Especificidad de la EspecieRESUMEN
After immunization of mice with the human hepatocellular carcinoma (HCC) cell line PLC/PRF/5, we produced monoclonal antibody KM-2, which allowed us to characterize a new HCC-associated antigen (KM-2 antigen) and to develop a sandwich-type radioimmunoassay. The KM-2 antigen was strongly expressed on the cell surface of HCC cell lines. Immunofluorescence staining of frozen sections of different tissues and tumors confirmed its specific expression on the cell surface of a group of HCC. The antigen was also detected in the bile canaliculi of normal liver. Its biochemical characterization revealed a high molecular weight (M(r) approximately 900,000) glycoprotein with an N-linked carbohydrate chain close to the peptide epitope recognized by the KM-2 monoclonal antibody. By the radioimmunoassay for the KM-2 antigen, the antigen was detected in sera of 72 (47%) of 154 patients with HCC and 3 (3%) of 102 patients with liver cirrhosis; it was not detected in 96 patients with chronic hepatitis or in 100 healthy control individuals. The positive rate of KM-2 antigen (72 of 154, 47%) was significantly (P less than 0.01) higher than that (51 of 154, 33%) of alpha-fetoprotein (AFP) when the cut-off level of AFP was taken as the widely accepted 400 ng/ml. No significant correlation was recognized between serum levels of the KM-2 antigen and AFP (r = 0.15; P greater than 0.05). In addition, among 103 patients with HCC whose AFP levels were less than 400 ng/ml, 31 (30%) were positive for the KM-2 antigen. Determination of the serum KM-2 antigen would be useful for the serodiagnosis of patients with HCC, particularly in cases with normal or low AFP levels.
Asunto(s)
Anticuerpos Monoclonales , Antígenos de Neoplasias/inmunología , Biomarcadores de Tumor/inmunología , Carcinoma Hepatocelular/diagnóstico , Neoplasias Hepáticas/diagnóstico , Anticuerpos Monoclonales/inmunología , Especificidad de Anticuerpos , Antígenos de Neoplasias/sangre , Técnica del Anticuerpo Fluorescente , Humanos , Técnicas In Vitro , Radioinmunoensayo , Células Tumorales Cultivadas , alfa-Fetoproteínas/metabolismoRESUMEN
We investigated the histological features of lymph nodes, focusing on monocytes/macrophages, in rhesus monkeys (Macaca mulatta) acutely infected with simian immunodeficiency virus (SIV). In monkeys infected with a pathogenic SIV, SIVmac239, MAC387(+) newly blood-derived macrophages markedly increased in number at paracortical areas at 11 to 14 days postinoculation, concomitant with the peak of the primary SIV antigenemia. The MAC387(+) macrophages densely gathered around high endothelial venules and formed cell clusters with CD3(+) T lymphocytes, tingible body macrophages, and plasmacytoid monocytes. In the cell clusters, CD3(+) T lymphocytes which closely adhered to the MAC387(+) macrophages enlarged in size, suggesting a histological manifestation of T-lymphocyte activation by macrophages. By 54 days postinoculation, when SIV antigenemia became undetectable, the MAC387(+) macrophages decreased in number and the cell cluster disappeared from paracortical areas. In contrast, the monkeys infected with a nef-deleted mutant of SIVmac239 showed lower levels of SIV antigenemia and lower numbers of MAC387(+) macrophages in paracortical areas than those infected with SIVmac239. These results indicate that MAC387(+) macrophages accumulate in paracortical areas for the period of the intense primary SIV antigenemia and may play an important role in activating naive T lymphocytes.
Asunto(s)
Ganglios Linfáticos/patología , Macrófagos , Síndrome de Inmunodeficiencia Adquirida del Simio/patología , Animales , Antígenos Virales/sangre , Agregación Celular , Productos del Gen gag/sangre , Hibridación in Situ , Cinética , Ganglios Linfáticos/inmunología , Macaca mulatta , Macrófagos/citología , Macrófagos/inmunología , Síndrome de Inmunodeficiencia Adquirida del Simio/inmunología , Síndrome de Inmunodeficiencia Adquirida del Simio/virología , Virus de la Inmunodeficiencia de los Simios/aislamiento & purificaciónRESUMEN
We present a long-surviving patient with Wiskott-Aldrich syndrome complicated by atypical progressive multifocal leukoencephalopathy (PML). MRI showed multiple tiny spots of Gd-DTPA-enhanced lesions on the T1-weighted image. Pathologic findings for brain biopsy were patchy demyelinated vascularized lesions infiltrated by a surprising number of eosinophils. The presence of polyomavirus JC was confirmed by in situ hybridization and polymerase chain reaction. PML should be included in the differential diagnosis when Gd-DTPA-enhanced spotty lesions are present in the white matter, especially in patients who have a mild immunologic defect.
Asunto(s)
Leucoencefalopatía Multifocal Progresiva/complicaciones , Leucoencefalopatía Multifocal Progresiva/diagnóstico , Imagen por Resonancia Magnética , Síndrome de Wiskott-Aldrich/complicaciones , Adulto , Encéfalo/patología , Gadolinio DTPA , Humanos , Leucoencefalopatía Multifocal Progresiva/patología , Masculino , Compuestos Organometálicos , Ácido Pentético/análogos & derivados , Tomografía Computarizada por Rayos XRESUMEN
In chronic schizophrenic patients treated with phenothiazines (Chlorpromazine, Levomepromazine, Perphenazine) for long periods (average = 8 years), high density lipoprotein cholesterol (HDL-C) levels were significantly lower (P less than 0.001) compared with normal controls. The HDL subfractions showed that HDL3-C was significantly low (P less than 0.005) whereas HDL2-C was not. Both serum apo A-I and apo A-II levels were also low (P less than 0.005 and P less than 0.001, respectively) in schizophrenics treated with phenothiazines. The serum triglycerides (TG) level was significantly higher (P less than 0.05) in patients treated with phenothiazines than in controls. No significant differences in total cholesterol (TC), TG and HDL-C were found between users and nonusers of benzodiazepine in schizophrenic patients receiving phenothiazines. In addition to chronic schizophrenic patients, 8 new patients with schizophrenia and related diseases were studied. The serum HDL-C level decreased by 24% within 1 week following administration of phenothiazines. No significant differences were found in TC and TG levels for 10 weeks after initiation of phenothiazine administration.
Asunto(s)
Antipsicóticos/uso terapéutico , Colesterol/sangre , Lipoproteínas HDL/sangre , Esquizofrenia/tratamiento farmacológico , Adulto , Ansiolíticos/uso terapéutico , Apolipoproteína A-I , Apolipoproteína A-II , Apolipoproteínas/sangre , Benzodiazepinas , Clorpromazina/uso terapéutico , HDL-Colesterol , Humanos , Masculino , Metotrimeprazina/uso terapéutico , Persona de Mediana Edad , Perfenazina/uso terapéutico , Esquizofrenia/sangre , Factores de Tiempo , Triglicéridos/sangreRESUMEN
To determine if there are seasonal variations in serum high density lipoprotein cholesterol (HDL-C), the concentration of HDL-C was measured monthly for 12 consecutive months in 31 healthy men and 24 male inpatients with schizophrenia. In addition to HDL-C, total cholesterol (TC) and triglyceride (TG) concentrations in serum were assayed, and low density lipoprotein cholesterol (LDL-C) was estimated by calculation. Mean serum HDL-C levels of schizophrenic patients were significantly low compared with those of healthy controls, 35 +/- 12 and 49 +/- 11 mg/dl, respectively. The TC levels of schizophrenic patients were significantly higher in January and March as compared with August. The HDL-C levels in summer and autumn were significantly lower than those in winter and spring in both healthy men and schizophrenic patients. The concentration of LDL-C was significantly high in September and October as compared with April in healthy men. In patients with schizophrenia, LDL-C level seemed higher in January and March as compared with August.
Asunto(s)
Colesterol/sangre , Lipoproteínas HDL/sangre , Esquizofrenia/sangre , Estaciones del Año , Adulto , HDL-Colesterol , LDL-Colesterol , Humanos , Lipoproteínas LDL/sangre , Masculino , Persona de Mediana Edad , Triglicéridos/sangreRESUMEN
The lectin amaranthin, purified from the seeds of Amaranthus caudatus, has been shown to react specifically with the Gal beta 1,3GalNAc-alpha and the NeuAc alpha 2,3Gal beta 1,3GalNAc-alpha sequence which represent the T antigen and the cryptic T antigen, respectively. We report here the development of labeling techniques that apply amaranthin to stain paraffin sections from rat fetuses. Amaranthin staining was inhibited by pre-incubation of lectin-gold complexes with 10 mM Gal beta 1,3GalNAc-alpha-O-benzyl (synthetic T antigen) or 10 mM Gal beta 1,3GalNAc-alpha-O-aminophenylethyl-human serum albumin (T antigen neoglycoprotein), asialoglycophorin, asialofetuin, and asialomucin. The beta-elimination reaction also abolished the lectin staining demonstrating specificity for O-glycosidically linked structures. A comparison with monoclonal anti-T antigen antibody immunostaining demonstrated that amaranthin detects the T antigen and its cryptic form in tissue sections. Application of the galactose oxidase-Schiff sequence abolished amaranthin (and anti-T antibody) binding to the T antigen but not to its cryptic form, and therefore permitted their differentiation in tissue sections. Histochemical evidence was obtained indicating that amaranthin is a more specific anti-T reagent than peanut lectin. Data are presented that show the differential expression of the T antigen and the cryptic T antigen in organs and cells of rat fetuses late in gestation. Therefore, amaranthin can be used for histochemical detection of the T antigen and the cryptic T antigen, and facilitates discrimination between them.
Asunto(s)
Antígenos de Carbohidratos Asociados a Tumores , Disacáridos/análisis , Feto/análisis , Isoantígenos/análisis , Lectinas , Lectinas de Plantas , Trisacáridos/análisis , Animales , Anticuerpos Monoclonales , Asialoglicoproteínas , Secuencia de Carbohidratos , Feto/ultraestructura , Oro , Histocitoquímica , Datos de Secuencia Molecular , Especificidad de Órganos , Aglutinina de Mani , Ratas , Proteínas Inactivadoras de Ribosomas , Proteínas Inactivadoras de Ribosomas Tipo 1RESUMEN
The Maackia amurensis leukoagglutinin has been shown to react specifically with the Neu5Ac (alpha 2,3) Gal sequence of asparagine-linked complex type oligosaccharides. We report here the preparation of Maackia amurensis lectin-gold complexes and their application for light and electron microscopic detection of the Neu5 Ac (alpha 2,3) Gal sequence in various tissues. The use of the lectin directly gold labeled was superior to a two-step cytochemical affinity technique using a fetuin-gold complex. The Maackia amurensis lectin-gold staining was inhibited by pre-incubation of the lectin-gold complexes with 50 mM alpha 2,3 sialyllactose, whereas alpha 2,6 sialyllactose up to concentrations of 1 M had no effect, thus demonstrating the high specificity of the histochemical staining. In addition to N-glycanase-sensitive asparagine-linked oligosaccharides, beta-elimination-sensitive serine/threonine-linked oligosaccharides could be detected. Data are presented which show that cellular staining patterns obtained with Maackia amurensis lectin-gold complexes may differ from those with elderberry bark lectin-gold, which detects the Neu5 Ac (alpha 2,6) Gal/GalN Ac sequence. Electron microscopic double labeling for direct study of the differential distribution of the Neu5 Ac (alpha 2,3) Gal and Neu5 Ac (alpha 2,6) Gal sequences is reported. Therefore, the availability of two sialic acid binding lectins with different linkage specificity for histochemistry provides the first opportunity to study tissue and cell type expression of these terminal sequences of glycoproteins.
Asunto(s)
Acetilglucosamina/metabolismo , Glucosamina/análogos & derivados , Fitohemaglutininas/metabolismo , Ácidos Siálicos/metabolismo , Acetilglucosamina/análisis , Aminoácidos/análisis , Animales , Conformación de Carbohidratos , Bovinos , Colon/citología , Colon/metabolismo , Colon/ultraestructura , Oro/metabolismo , Histocitoquímica/métodos , Microscopía Electrónica , Páncreas/citología , Páncreas/metabolismo , Páncreas/ultraestructura , Lectinas de Plantas , Ratas , Ácidos Siálicos/análisis , Coloración y Etiquetado , Glándula Submandibular/citología , Glándula Submandibular/metabolismo , Glándula Submandibular/ultraestructura , Porcinos , ÁrbolesRESUMEN
In view of the strong association between the acquired immunodeficiency syndrome (AIDS) and sexually transmitted diseases (STDs), we screened 182 human immunodeficiency virus (HIV)-1 infected patients over a 15-month period for serological markers to previously encountered or current STDs, most of viral etiology. The relationship between their immunological and clinical status and the prevalence of STDs was assessed and compared with that of 88 HIV-seronegative patients. Hepatitis B virus and Treponema pallidum were the most frequently occurring pathogens in both HIV-1-infected and HIV-seronegative patients. Hepatitis C virus (HCV) infection was also observed in both groups, but no HIV-seronegative patient was infected with human T-lymphotropic virus type 1 (HTLV-1). The Centers for Disease Control clinical staging of A1 through C3, representing asymptomatic to severe AIDS conditions, was observed in HIV-1 patients with or without STDs. A mean CD4 count of 288 cells per microliter (95% CI of 237-340 cells per microliter) in HIV-1 patients was significantly lower (P < 0.05) than that in HIV-seronegative individuals with 1019 cells per microliter (95% CI of 924-1115 cells per microliter), irrespective of whether subjects in either group had previous or current STDs. The mean CD4 count of patients with a single infection from HIV-1 was not significantly different (P = 0.36) from that of HIV-1 patients with multiple infections. HIV-1 infection alone appears to be responsible for the marked immunodeficiency status of seropositive patients observed in this study.
Asunto(s)
Infecciones Oportunistas Relacionadas con el SIDA/inmunología , VIH-1 , Enfermedades de Transmisión Sexual/inmunología , Infecciones Oportunistas Relacionadas con el SIDA/epidemiología , Infecciones Oportunistas Relacionadas con el SIDA/fisiopatología , Adulto , Recuento de Linfocito CD4 , Femenino , Ghana/epidemiología , Seropositividad para VIH/complicaciones , Seropositividad para VIH/inmunología , Seropositividad para VIH/fisiopatología , Humanos , Masculino , Cómputos Matemáticos , Persona de Mediana Edad , Prevalencia , Enfermedades de Transmisión Sexual/epidemiología , Enfermedades de Transmisión Sexual/fisiopatologíaRESUMEN
We have used a particle agglutination (PA) test, Western blot (WB) test, polymerase chain reaction (PCR) test, and virus isolation to define the human immunodeficiency virus (HIV) status of 17 acquired immunodeficiency syndrome (AIDS), 6 AIDS-related complex (ARC), and 2 asymptomatic Ghanaians. HIV-1 antibodies were more frequently detected. The PCR detected 66.7% HIV-1, 11.1% HIV-2, and 5.6% of both HIV-1 and HIV-2 proviral DNA in peripheral blood mononuclear cell (PBMCs) and PBMC-Molt 4 coculture samples tested. Of the 12 viruses isolated from the 25 Ghanaians, 9 were HIV-1, 2 were HIV-2, and both HIV-1 and HIV-2 were isolated from 1 individual. Two of the HIV-1 isolates were from ARC patients who have been PA negative and either HIV-1 or HIV-2 WB indeterminate for more than 1 year without developing antibodies to HIV envelope proteins. Our results indicate that HIV-1 is now predominant in Ghanaian AIDS and ARC patients and that dual infection can occur.
PIP: While HIV is believed to be the causative agent for AIDS, many clinically diagnosed AIDS and AIDS-related complex (ARC) cases in Ghana have been reported to be negative or indeterminate for HIV antibodies. Dual seropositive reactions have also been common among AIDS and ARC cases in the country. A particle agglutination (PA) test, Western blot (WB), polymerase chain reaction (PCR), and virus isolation were used to define the HIV status of 17 AIDS, 6 ARC, and 2 asymptomatic Ghanaians. The PA test detected HIV-1 antibodies in 72% of the plasma samples, 94.4% of which were also positive according to WB. 1 sample was indeterminate by WB and 2 HIV-1 negative samples were determined to be positive by WB. HIV-2 was detected by PA in 32% of all samples, of which 87.5% were confirmed by WB. PCR detected 66.7% of HIV-1 cases, 11.1% of HIV-2, and 5.6% of both HIV-1 and HIV-2 proviral DNA in peripheral blood mononuclear cells (PBMCs) and PBMC-Molt 4 coculture samples tested. 12 viruses were isolated from the 25 subjects; 9 were identified as HIV-1, 2 as HIV-2, and 1 person was infected with both HIV-1 and HIV-2. 2 of the HIV-1 isolates were from ARC patients who had been PA-negative and either HIV-1 or HIV-2 WB indeterminate for more than 1 year without developing antibodies to HIV envelope proteins.
Asunto(s)
Complejo Relacionado con el SIDA/virología , Síndrome de Inmunodeficiencia Adquirida/virología , VIH-1/aislamiento & purificación , VIH-2/aislamiento & purificación , Complejo Relacionado con el SIDA/epidemiología , Síndrome de Inmunodeficiencia Adquirida/epidemiología , Pruebas de Aglutinación , Secuencia de Bases , Western Blotting , Cartilla de ADN , Ghana/epidemiología , Humanos , Datos de Secuencia Molecular , Reacción en Cadena de la PolimerasaRESUMEN
We have isolated a replication-competent, full-length molecular clone of HIV-1 CRF02_AG, designated p97GH-AG1, by reconstituting two separately amplified genomic regions of an HIV-1 provirus of a 1997 Ghanaian isolate. The phylogenetic and recombination breakpoint analyses revealed that 97GH-AG1 had an A/G recombinant structure similar to that of prototype Nigerian isolate IbNG. The 17-nucleotide insertion downstream of the primer-binding site appeared to be a common sequence signature specific to most CRF02_AG strains, including 97GH-AG1. 97GH-AG1 showed an R5 phenotype and exerted productive infection in both HOS and NP2 cell infectivity assays, whereas it failed to show a detectable level of progeny production in peripheral blood mononuclear cells (PBMCs). The data may suggest the presence of unknown determinant(s) that dictate efficient replication in PBMCs, but that are not required for replication in immortalized cell lines.
Asunto(s)
Seropositividad para VIH/virología , VIH-1/clasificación , VIH-1/genética , Filogenia , Recombinación Genética , Replicación Viral , Secuencia de Aminoácidos , Secuencia de Bases , ADN Viral , Femenino , Ghana , Humanos , Persona de Mediana Edad , Datos de Secuencia Molecular , Mapeo Nucleótido , Reacción en Cadena de la PolimerasaRESUMEN
The phylogenetic variability of part of the long terminal repeat (LTR) region of HIV-2 strains isolated in 1995 from five individuals residing in Bissau, the capital city of Guinea-Bissau, and collected from seven persons from Kumasi, Ghana in 1996-1997, was analyzed. All Guinean samples and all but one Ghanaian sample clustered with HIV-2 subtype A. One Ghanaian sample (14%) was classified as HIV-2 subtype B. This study adds to previous reports on HIV-2 subtype distribution in West Africa indicating local prevalence of HIV-2 subtype B in Ivory Coast and neighboring Ghana.
Asunto(s)
Infecciones por VIH/epidemiología , Duplicado del Terminal Largo de VIH/genética , VIH-2/clasificación , ADN Viral/análisis , Ghana/epidemiología , Guinea Bissau/epidemiología , Infecciones por VIH/virología , VIH-2/genética , Humanos , Datos de Secuencia Molecular , Filogenia , Análisis de Secuencia de ADNRESUMEN
PIP: The authors examined HIV-1 genetic variation among 19 HIV-1-infected people of mean age 34.5 years living in Accra, Akwatia, Kumasi, and Ho, in Ghana. One person was of unknown origin. Blood samples were collected between December 1993 and January 1996. 16 of the HIV-1 specimens clustered with members of subtype A, but the clustering was not supported by 70% or more of the bootstrap tests. Two samples clustered with subtype D strains, supported by 92.5% of the bootstrap trees, and one sample clustered with subtype G strains, supported by 96.2% of the bootstrap trees. For the Ghanaian specimens belonging to subtype A, interhost distances at the nucleotide level averaged 14.9%, of range 7.83-20.9%. The interhost distance between the two subtype D samples was 8.2%. A cocirculation of subtypes A, D, and G was identified in Akwatia.^ieng