RESUMEN
OBJECTIVE: Cytomegalovirus (CMV) is the leading cause of viral congenital infection. Some viral factors have been proposed to be CMV pathogenicity markers. The objective of this study was to investigate the frequency of congenital CMV infection in symptomatic patients and the possible association with the CMV glycoprotein B (gB) genotype and viral load. STUDY DESIGN: A total of 361 newborns (NB) and 158 pregnant women (PW) with clinically suspected CMV infection were enrolled. Studied samples included urine, saliva, serum, vaginal swabs and amniotic fluid. CMV infection was diagnosed by multiplex nested PCR. CMV gB genotyping was performed on infected samples, followed by viral load determination. RESULTS: Overall, 18.7% of the tested patients were positive for CMV infection, 19.7% of NB were congenitally infected and 16.5% of PW showed active CMV infection. gB-2 was the most prevalent genotype detected (39/97 patients). gB CMV mixed infections were detected in 12 patients. gB-2 was associated with mono-infections (P<0.01). Mixed infections showed higher levels of viral load compared with gB mono-infection (P=0.03). Hepatomegaly, splenomegaly, jaundice, sepsis-like syndrome and malformations were the most prevalent clinical findings. gB-4 was more frequently associated with sepsis-like syndrome than other gB genotypes (P=0.04, odds ratio=4.3, confidence interval: 0.9 to 21.6). The difference in medians of CMV load was statistically significant among patients presenting different clinical signs (P=0.04). CONCLUSIONS: This study showed that CMV is a frequent cause of congenital infection in symptomatic Cuban patients. Despite gB2 being the most frequently detected, gB-4 was the only genotype associated with clinical features (sepsis-like syndrome in NB). No other associations among specific genotypes and clinical characteristics were found. Further studies are needed to clarify the role that viral load and genotype play in the outcome of congenital infection.
Asunto(s)
Infecciones por Citomegalovirus/congénito , Infecciones por Citomegalovirus/diagnóstico , Citomegalovirus/genética , Proteínas del Envoltorio Viral/genética , Carga Viral , Biomarcadores/análisis , Cuba/epidemiología , Infecciones por Citomegalovirus/epidemiología , Infecciones por Citomegalovirus/virología , ADN Viral/análisis , Femenino , Genotipo , Humanos , Recién Nacido , Reacción en Cadena de la Polimerasa Multiplex/métodos , Embarazo , Estadísticas no Paramétricas , Ultrasonografía PrenatalRESUMEN
Retained low-density lipoproteins (LDL) by arterial glycosaminoglycans (GAG) are more susceptible to reactive oxygen species-mediated oxidation, contributing to oxidative stress and atherosclerosis. Recently, we reported the properties of the chimeric mouse/human monoclonal antibody chP3R99-LALA to bind sulfated GAG, to inhibit LDL-chondroitin sulfate binding, and to avoid LDL oxidation in vitro. Here, we hypothesized that chP3R99-LALA treatment might reduce aortic oxidative stress in a therapeutic setting. Redox biomarkers and serum lipids were determined by spectrophotometric methods. Subcutaneous administration of five doses (100 µg) of chP3R99-LALA, after Lipofundin administration (2 mL/kg/day, i.v.) during 8 days, reduced atherosclerotic lesion development, which was not associated with a serum lipid modulation. In contrast, the treatment with chP3R99-LALA reduced (p < 0.05) malondialdehyde and protein oxidation, induced a restoration of reduced glutathione level, of the superoxide dismutase and catalase activities and of endothelial nitric oxide level. Thus, the antiatherogenic effect of chP3R99-LALA treatment seems to be associated with a reduction of aortic oxidative stress. These results contribute in understanding the molecular mechanisms associated with chP3R99-LALA atheroprotection and support the use of anti-GAG antibody-based immunotherapy as a potential tool to treat the atherosclerosis.
Asunto(s)
Anticuerpos Antiidiotipos/administración & dosificación , Aorta/efectos de los fármacos , Aterosclerosis/inmunología , Glicosaminoglicanos/inmunología , Animales , Aorta/inmunología , Aorta/patología , Aterosclerosis/tratamiento farmacológico , Aterosclerosis/metabolismo , Modelos Animales de Enfermedad , Glicosaminoglicanos/metabolismo , Humanos , Lipoproteínas LDL/inmunología , Lipoproteínas LDL/metabolismo , Estrés Oxidativo/efectos de los fármacos , Conejos , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Probiotics are microorganisms that have demonstrated beneficial effects on human health. Probiotics are usually isolated from the commensal microflora that inhabits the skin and mucosas. We propose that probiotics represent the species of microorganisms that have established a symbiotic relationship with humans for the longest time. Cultural practices of ancient human societies used to favor that symbiosis and the transmission of probiotics from generation to generation. New practices, introduced as a result of industrialization, such as childbirth by surgical delivery, ingestion of pasteurized and synthetic compounds-supplemented food, cleaner homes, indiscriminate use of antibiotics and so on, have led in recent years to the replacement of probiotics by other microorganisms that are not as well adapted to the microenvironments of the human body. These newly settled microorganisms lack many of the beneficial effects of probiotics. Our hypothesis is that the sudden change (from an evolutive perspective) in human intestinal microflora may importantly contribute to the rise in the incidence of autoimmune diseases, observed in the last half a century.
Asunto(s)
Autoinmunidad , Probióticos/uso terapéutico , Humanos , Recién Nacido , Mucosa Intestinal/microbiología , Modelos Biológicos , Modelos Inmunológicos , SimbiosisRESUMEN
A cross-sectional study was performed in 60 Cuban women of child-bearing age who were seropositive for human immunodeficiency virus (HIV) and 60 controls. Human papillomavirus (HPV) was identified most frequently, with oncogenic HPV serotypes 16, 33 and 58 detected in HIV-positive patients, and serotypes 11, 33 and 51 in the controls (relative risk 4.41; 95% CI 2.21-8.29). Syphilis and hepatitis B and C viruses were detected exclusively in HIV-seropositive women (p<0.05). Sexually transmitted diseases (STDs) appeared to pose a substantial health problem, especially for HIV-positive women. Clinics should consider screening and treatment for STDs as part of their HIV prevention programmes.
Asunto(s)
Papillomaviridae , Enfermedades de Transmisión Sexual/epidemiología , Adolescente , Adulto , Comorbilidad , Estudios Transversales , Cuba/epidemiología , Femenino , Seropositividad para VIH/epidemiología , Hepatitis B/epidemiología , Hepatitis C/epidemiología , Humanos , Persona de Mediana Edad , Infecciones por Papillomavirus/epidemiología , Prevalencia , Sífilis/epidemiologíaRESUMEN
We have expressed a recombinant Dengue 4 virus envelope glycoprotein (E4rec), truncated at its C-terminus by 53 amino acids, in Pichia pastoris. The presence of E4rec was confirmed by Western-blot using anti-DEN 4 hyper immune mouse ascitic fluid. E4rec migrated during SDS-PAGE as a 64 kDa protein. Treatment with endoglycosidases showed that the E protein was modified by the addition of short mannose chains and the absence of hyperglycosylation. When administered to BALB-C mice, E4rec elicited a DEN 4 neutralizing antibody response haemagglutination inhibition antibodies and specific memory T cell response. Mice immunized were also significantly protected against lethal DEN 4 virus challenge (86.6%, p < 0.001).
Asunto(s)
Anticuerpos Antivirales/sangre , Virus del Dengue/inmunología , Vacunas Sintéticas/inmunología , Proteínas del Envoltorio Viral/inmunología , Vacunas Virales/inmunología , Animales , Dengue/prevención & control , Inmunización , Ratones , Ratones Endogámicos BALB C , Pruebas de Neutralización , Pichia/genética , Proteínas Recombinantes/inmunologíaAsunto(s)
ADN Viral/genética , Neoplasias Laríngeas/virología , Neoplasias de Células Escamosas/virología , Papillomaviridae/genética , Papillomaviridae/aislamiento & purificación , Infecciones por Papillomavirus/diagnóstico , Reacción en Cadena de la Polimerasa , Infecciones Tumorales por Virus/diagnóstico , Adulto , Formaldehído , Humanos , Neoplasias Laríngeas/diagnóstico , Persona de Mediana Edad , Neoplasias de Células Escamosas/diagnósticoRESUMEN
The polymerase chain reaction technique was applied to detect sequences of human Papillomavirus (HPV) by controls of cellular lines of cervical cancer and of tissues obtained through biopsy with a HPV-positive clinical diagnosis. A set of consensus oligonucleotides, which are complementary to a highly conserved region within the open reading frame E1 of the viral genome of HPV affecting the cervical mucosa, was used. With these primers it was possible to amplify DNA sequences corresponding to HPV 6 and 11, considered in the low risk group, and to HPV 16, 18, 31 and 33, included in the high risk group. The study of the sensitivity of the amplification technique showed a level of detection of 3,5 viral particles per each cellular diploid genome.
Asunto(s)
Papillomaviridae/genética , Reacción en Cadena de la Polimerasa/métodos , ADN Viral/análisis , ADN Viral/genética , ADN Viral/aislamiento & purificación , Genoma Viral , Humanos , Sondas de Oligonucleótidos , Mapeo Restrictivo , Sensibilidad y Especificidad , Análisis de Secuencia de ADN , Células Tumorales Cultivadas , Cultivo de Virus/métodosRESUMEN
Wistar rats were lesioned into the nigrostriatal pathway with 6-OHDA. The D-amphetamine-induced circling behavior test was performed to evaluated lesion efficiency. Animals that showed more than 620 turns/90 min were named totally lesioned animals (TLA). The group of rats that performed less than 620 turns/90 min were named partially lesioned animals (PLA). The contents of DA and its catabolites in the striata of these groups, and in the same tissue of the untreated animals, were measured. Moreover, the striatal glutathione-S-transferase (GST) specific activity for all groups was tested, and the kinetics parameters for GST purified from the whole brain were evaluated from other three similar groups. The striatal DA depletion on TLA was greater than in PLA. Striatal GST activity showed a significantly bilateral increase in PLA, whereas TLA exhibited only and ipsilateral augment. There were also differences between groups about the kinetic parameters of the purified brain enzyme. The possible role of GST on the interindividual lesion response difference was analyzed.
Asunto(s)
Cuerpo Estriado/efectos de los fármacos , Glutatión Transferasa/biosíntesis , Neuronas/efectos de los fármacos , Sustancia Negra/efectos de los fármacos , Ácido 3,4-Dihidroxifenilacético/metabolismo , Animales , Cuerpo Estriado/citología , Cuerpo Estriado/enzimología , Dopamina/metabolismo , Inducción Enzimática , Ácido Homovanílico/metabolismo , Masculino , Neuronas/enzimología , Neurotoxinas , Oxidopamina , Ratas , Ratas Wistar , Sustancia Negra/citología , Sustancia Negra/enzimologíaRESUMEN
The aim of this study was to develop a polymerase chain reaction (PCR) for the detection of respiratory syncytial virus (RSV) genomes. The primers were designed from published sequences and selected from conserved regions of the genome encoding for the N protein of subgroups A and B of RSV. PCR was applied to 20 specimens from children admitted to the respiratory ward of "William Soler" Pediatric Hospital in Havana City with a clinical diagnosis of bronchiolitis. The PCR was compared with viral isolation and with an indirect immunofluorescence technique that employs monoclonal antibodies of subgroups A and B. Of 20 nasopharyngeal exudates, 10 were found positive by the three assayed methods. In only two cases, samples that yielded positive RNA-PCR were found negative by indirect immunofluorescence and cell culture. Considering viral isolation as the "gold standard" technique, RNA-PCR had 100% sensitivity and 80% specificity. RNA-PCR is a specific and sensitive technique for the detection of the RSV genome. Technical advantages are discussed.
Asunto(s)
Reacción en Cadena de la Polimerasa , Infecciones por Virus Sincitial Respiratorio/diagnóstico , Virus Sincitial Respiratorio Humano/genética , Humanos , ADN Polimerasa Dirigida por ARN , Mapeo RestrictivoRESUMEN
The immune response of a group of patients with epidemical neuropathy and of controls was studied by the immunoblotting technique against proteins of the Coxsackie virus and the proteins of slow effect isolated in our laboratory. 13 sera of patients with epidemical neuropathy and 9 sera of controls were studied. Of the 13 sera studied, 8 (61.5%) recognized protein VPI and 2 sera (15.3%) protein VP0 of the strain 47.93. Of the 9 controls studied, 4 (44.4%) recognized protein VPI and 3 (33.3%) protein VP0 only. With the antigen prepared from the slow effect strain it was obtained a specific signal in 5 (38.5%) sera of patients and in 2 sera (22.5%) of controls. It should be stressed that in this last case the protein observed had a molecular weight of 41,300 D, and that its size was smaller than that of the preceding protein detected against the strain 47.93 was of 45,000 D.
Asunto(s)
Anticuerpos Antivirales/sangre , Infecciones por Coxsackievirus/inmunología , Brotes de Enfermedades , Enterovirus/inmunología , Enfermedades del Nervio Óptico/inmunología , Enfermedades del Sistema Nervioso Periférico/inmunología , Proteínas Virales/inmunología , Animales , Especificidad de Anticuerpos , Western Blotting , Chlorocebus aethiops , Infecciones por Coxsackievirus/epidemiología , Infecciones por Coxsackievirus/virología , Cuba/epidemiología , Enterovirus/clasificación , Enterovirus/aislamiento & purificación , Humanos , Peso Molecular , Enfermedades del Nervio Óptico/sangre , Enfermedades del Nervio Óptico/epidemiología , Enfermedades del Nervio Óptico/virología , Enfermedades del Sistema Nervioso Periférico/sangre , Enfermedades del Sistema Nervioso Periférico/epidemiología , Enfermedades del Sistema Nervioso Periférico/virología , Conejos , Estudios Seroepidemiológicos , Células Vero , Proteínas Virales/químicaRESUMEN
A study of conjunctivitis cases with a possible viral cause was carried out in Havana City during June and August, 1993. Thirty seven cases from "Ramón Pando Ferrer" Ophthalmological Hospital were studied, of whom 13 isolations (35.1%) in cell culture of Hela cells were obtained. Isolations showed a cytopathic effect characteristic of Adenovirus. A direct immunofluorescence with conjugated serum of rabbit specific to Adenovirus was performed and results were positive in 100% of cases. A DNA restriction analysis of each isolated was performed for their classification. Restriction endonucleases SmaI and Hind III were used and 2 isolates were classified as Adenovirus type 3, and 6 as Adenovirus type 7b, 4 out of 13 isolates were not classified because of the degradation of the genetic material.