Antiviral activity of the natural alkaloid anisomycin against dengue and Zika viruses.

Flaviviruses constitute a public health concern because of their global burden and the lack of specific antiviral treatment. Here we investigated the antiviral activity of the alkaloid anisomycin against dengue (DENV) and Zika (ZIKV) viruses. At non-cytotoxic concentrations, anisomycin strongly inhibited the replication of reference strains and clinical isolates of all DENV serotypes and Asian and African strains of ZIKV in Vero cells. Anisomycin also prevented DENV and ZIKV multiplication in human cell lines. While initial steps of DENV and ZIKV replicative cycle were unaffected, a high inhibition of viral protein expression was demonstrated after treatment with anisomycin. DENV RNA synthesis was strongly reduced in anisomycin treated cultures, but the compound did not exert a direct inhibitory effect on 2' O-methyltransferase or RNA polymerase activities of DENV NS5 protein. Furthermore, anisomycin-mediated activation of p38 signaling was not related to the antiviral action of the compound. The evaluation of anisomycin efficacy in a mouse model of ZIKV morbidity and mortality revealed that animals treated with a low dose of anisomycin exhibited a significant reduction in viremia levels and died significantly later than the control group. This protective effect was lost at higher doses, though. In conclusion, anisomycin is a potent and selective in vitro inhibitor of DENV and ZIKV that impairs a post-entry step of viral replication; and a low-dose anisomycin treatment may provide some minimal benefit in a mouse model.

Limitación del esfuerzo terapéutico en pacientes hospitalizados en el Servicio de Medicina Interna / Withholding and withdrawing treatment in patients admitted in an Internal Medicine ward

Introducción. Muchos pacientes ingresados en Medicina Interna presentan mala calidad de vida y/o mal pronóstico vital, por lo que son potenciales candidatos a limitación del esfuerzo terapéutico (LET). El objetivo primario consiste en describir qué medidas de LET se realizan en los pacientes fallecidos en un servicio de Medicina Interna de un hospital de Madrid. El secundario, la descripción del tipo de paciente en el que se realiza LET. Material y métodos. Estudio observacional durante 6 meses de 2011 y 2012. Se incluyeron todos los pacientes fallecidos en Medicina Interna. Resultados. Ingresaron 2.007 pacientes y fallecieron 211 (10,5%). Edad media 85 ± 9 años, el 57% (121) mujeres. Peso medio de los fallecidos 3,4. Ciento tres de los fallecidos (48,8%) provenía de residencia y 105 (49,8%) fueron considerados enfermos terminales. En 182 (86,3%; IC 95%: 81,4-91,1) se estableció durante el ingreso alguna orden de LET, en 99 (46,9%; IC 95%: 39,9-53,9) 2, y en 31 (14,7%; IC 95%: 9,6-19,7) 3 o más. Órdenes de LET más pautadas: «orden de no reanimación cardiopulmonar (154; 73%), no realización de «maniobras agresivas» (80; 38%), rechazo de antibioterapia (19; 9%), no ingreso en UCI (18; 8,5%) y no realización de cirugía (11; 5,2%). Conclusiones. La LET es una estrategia muy frecuente en los pacientes que fallecen en Medicina Interna. Las más frecuentes son la «orden de no reanimación cardiopulmonar» y la no realización de «medidas agresivas». Dichas órdenes se indican en pacientes de edad avanzada, con importante comorbilidad y alta incidencia de demencia y enfermedad terminal (AU)

Introduction. Many of the patients admitted to a general medical ward have a compromised quality of life, or short life expectancy, so they are potential candidates for withhold/withdraw (WH/WD) treatment. The first objectif was to describe which measures were WH/WD among patients who died during their admission in a general medical ward from a tertiary hospital in Madrid. Secondly, to define the clinical characteristics of this population. Material and methods. A cross-sectional descriptive study during 6 months from 2011 and 2012 of all the patients dead while their admission in the Internal Medicine Department. Results. 2007 patients were admitted, 211 died (10.5%). 121 (57%) were female, with 85 ± 9 years of mean age. 103 (48.8%) came from a residential facility and 105 fulfilled terminality criteria (49.8%). One decision to WH/WD treatment was made in 182 patients (86.3%, CI 95%: 81.4-91.1), two in 99 cases (46.9%, CI 95%: 39.9-53.9) and 3 or more in 31 subjects (14.7%, CI 95%: 9.6-19.7). The most frequent decisions involved do-not-resuscitate orders (154, 73.0%), rejection of «aggressive treatment measures» (80, 38.0%), use of antibiotics (19, 9.0%), admission in ICU (18, 8.5%), and/or surgical treatment (11, 5.2%). Conclusions. WH/WD treatment is very frequent among patients who died in a general medical ward. The most frequent involved do-not-resuscitate orders and rejection of «aggressive treatment measures». WH/WD decisions are adopted in an elderly population, with extensive comorbidity and an elevated prevalence of advanced dementia and/or terminal disease (AU)

[Withholding and withdrawing treatment in patients admitted in an Internal Medicine ward]. / Limitación del esfuerzo terapéutico en pacientes hospitalizados en el Servicio de Medicina Interna.

INTRODUCTION: Many of the patients admitted to a general medical ward have a compromised quality of life, or short life expectancy, so they are potential candidates for withhold/withdraw (WH/WD) treatment. The first objectif was to describe which measures were WH/WD among patients who died during their admission in a general medical ward from a tertiary hospital in Madrid. Secondly, to define the clinical characteristics of this population. MATERIAL AND METHODS: A cross-sectional descriptive study during 6 months from 2011 and 2012 of all the patients dead while their admission in the Internal Medicine Department. RESULTS: 2007 patients were admitted, 211 died (10.5%). 121 (57%) were female, with 85±9 years of mean age. 103 (48.8%) came from a residential facility and 105 fulfilled terminality criteria (49.8%). One decision to WH/WD treatment was made in 182 patients (86.3%, CI 95%: 81.4-91.1), two in 99 cases (46.9%, CI 95%: 39.9-53.9) and 3 or more in 31 subjects (14.7%, CI 95%: 9.6-19.7). The most frequent decisions involved do-not-resuscitate orders (154, 73.0%), rejection of «aggressive treatment measures¼ (80, 38.0%), use of antibiotics (19, 9.0%), admission in ICU (18, 8.5%), and/or surgical treatment (11, 5.2%). CONCLUSIONS: WH/WD treatment is very frequent among patients who died in a general medical ward. The most frequent involved do-not-resuscitate orders and rejection of «aggressive treatment measures¼. WH/WD decisions are adopted in an elderly population, with extensive comorbidity and an elevated prevalence of advanced dementia and/or terminal disease.

Antiherpes simplex virus type 2 activity of the antimicrobial peptide subtilosin.

AIMS: In this study, we evaluated the antiviral activity of subtilosin, a cyclical peptide isolated from Bacillus amyloliquefaciens, against herpes simplex virus type 2 (HSV-2) in cell cultures and we investigated subtilosin mode of action. METHODS AND RESULTS: We determined, using a virus yield inhibition assay, that noncytotoxic concentrations of subtilosin inhibit HSV-2 replication in Vero cell cultures. Subtilosin strongly inhibited extracellular and total virus production even when it was added at 8 h postinfection indicating that not only virus release but also viral particle formation is impeded by the antiviral peptide. Although viral glycoprotein gD level of expression is not affected by the bacteriocin, an altered pattern of gD intracellular localization was detected by immunofluorescence assay in subtilosin-treated culture. On the other hand, at high concentrations, subtilosin displays virucidal action. CONCLUSIONS: Subtilosin displays antiviral and virucidal actions against HSV-2. The target of subtilosin inhibitory effect would be late stages of the viral replicative cycle such as viral glycoprotein intracellular transport. SIGNIFICANCE AND IMPACT OF THE STUDY: Given its antimicrobial activity and its safety for human tissues, subtilosin could represent a valuable alternative to be considered in the development of new microbicide formulations.

Plant and animal steroids a new hope to search for antiviral agents.

Scientific literature provides evidence about the use of steroids as an adjunct treatment to antiviral therapies. Immunomodulatory activity of some steroids would account for the recovery in patients with herpetic and other viral infections. However, in vitro studies have demonstrated a direct antiviral effect of this kind of molecules. In this review we discuss recent reports about the mechanism of antiviral action of steroids from animal and plant origin. Chemical structures of most active compounds are also provided.

The changing pattern of tuberculosis and HIV co-infection in immigrants and Spaniards in the last 20 years.

OBJECTIVES: To evaluate the impact of immigration on tuberculosis (TB)-HIV co-infection in Spain in a prospective cohort of HIV patients. METHODS: Among 7761 HIV patients, we evaluated 1284 with at least one episode of TB between 1987 and 2006. Variables were compared between immigrants and Spaniards. RESULTS: Incidence of TB decreased from 20 to five cases per 100 patient-years in 2006 (P<0.01) and was always higher in immigrants than in Spaniards. The proportion of immigrants increased, reaching almost 50% of both new cases of HIV and TB-HIV co-infection in 2006. In 34.4% of patients, TB and HIV infection were diagnosed within the same year; simultaneous diagnosis was more frequent in immigrants (83.3%vs. 16.7%, P<0.001). Mortality was associated independently with age [hazard ratio (HR) 1.03, 95% confidence interval (CI) 1.01-1.05], TB diagnosis before 1996 (HR 2.6, 95% CI 1.8-3.6), use of highly active antiretroviral treatment (HR 0.494, 95% CI 0.37-0.66) and CD4 cell count at TB diagnosis (HR 0.996, 95% CI 0.995-0.997). CONCLUSIONS: Immigrants have a major impact on the incidence of TB in HIV patients, slowing down the decreasing trend in Spain. Simultaneous diagnosis of the co-infection in immigrants reveals a need to intensify HIV case finding in immigrants in Spain.

Economic evaluation of assistance to HIV patients in a Spanish hospital.

BACKGROUND: Little is known about the global effects of HAART on the use of medical resources after the complete implementation of this therapy in Spain. This study was designed to determine the use of medical resources and the costs of health care for HIV-infected patients. METHODS: All patients with HIV infection who came to our institution during the year 2002 were included in the study. We analyzed the global assistance data and pharmaceutical costs during the year. Costs were calculated based on a unitary cost for DRG and an officially assigned standard cost for outpatient clinic, visits to the day care unit and to the emergency room (ER), outpatient surgery, and total costs of pharmacy. RESULTS: The total cost for HIV-related health care assistance was euro739,048. The cost related to admissions was euro150,766.60; euro8631 per first visit and euro49,199.40 per successive visit; euro5085.10 per day care unit; euro14,920 per outpatient surgery; euro7655.70 per ER visit; and euro491,342.40 per antiretroviral treatment. A significant proportion of the total outpatient assistance was given by physicians other than HIV specialists, namely, 63% of the costs attributed to the first visit and 41% per successive visit. CONCLUSION: More than 50% of the costs of caring for HIV-infected patients are still attributed to antiretroviral therapy. Specialists other than infectious disease specialists provide a significant proportion of outpatient assistance. A method to control HIV costs is greatly needed.

A randomized controlled trial investigating the efficacy and safety of switching from a protease inhibitor to nevirapine in patients with undetectable viral load.

OBJECTIVE: To assess the antiviral efficacy and safety of switching from a protease inhibitor (PI) to nevirapine in patients with long-term HIV-1 RNA suppression on PI-containing regimens, and to assess its influence in the adherence to treatment. METHODS: In an open-label multicentre study, 160 HIV-infected patients with undetectable viral load for at least 6 months on a PI-containing regimen were randomized to either continue with their PI regimen (n=79) or replace PI with nevirapine (n=81). Clinical assessment included plasma HIV-1 RNA, blood chemistry, haematology, lymphocyte counts and adverse events reports. Adherence to treatment and lipodystrophy syndrome were assessed by patient self-reporting. RESULTS: Treatment efficacy was equivalent in the two arms, for patients with viral loads either above or below 100 000 HIV-1 RNA copies/mL. The increase in CD4 cell count was significant in both arms (P<0.00001) but the average CD4 cell count at 48 weeks was slightly higher in the nevirapine arm (596 vs. 569; P=0.1588). The number of patients with severe hypertriglyceridaemia (>400 mg/dL) after 48 weeks of treatment decreased in the nevirapine arm (from 11 to six), but increased in the PI arm (from four to 11) and led to treatment discontinuation in two patients. Lipodystrophy changes increased in 15% of patients in the PI arm but decreased in 4% of patients in the nevirapine arm. Finally, although adherence was similar in the two arms, patients reported that it required significantly less effort to stay on treatment in the nevirapine arm. Conclusions The results indicate that switching from PI to nevirapine is as effective as continuing with PI for maintaining viral control, even in patients with baseline viral load above 100,000 copies/mL. In addition, reductions in hypertriglyceridaemia and lipodystrophy and in the effort required to stay on treatment were observed.

Concentraciones plasmáticas de carotenos y vitaminas antioxidantes en personas de edad avanzada: influencia del tabaquismo. Proyecto HALE / Plasma concentrations of carotene and antioxidant vitamins in elderly people: the influence of smoking. HALE project

Introducción. Los carotenos y las vitaminas antioxidantes se asocian con un menor riesgo de aterosclerosis y enfermedad coronaria. Hay estudios que apoyan que el hábito tabáquico modifica el estado nutricional de estos micronutrientes. En este estudio se analizan las concentraciones plasmáticas de los carotenos y las vitaminas antioxidantes en varones y mujeres no fumadores, ex fumadores y fumadores. Pacientes y métodos. Se seleccionó aleatoriamente para participar en el estudio base (1988-1989) del proyecto SENECA (Survey in Europe on Nutrition and the Elderly, a Concerted Action), de la Unión Europea, a 2.200 habitantes de 19 ciudades europeas, Hamme (Bélgica), Roskilde (Dinamarca), Chateau Renault-Amboise, Haguenau y Romans (Francia), Markopoulo (Grecia), Anogia-Archanes (Creta), Monor (Hungría), Padua, Fara Sabina-Magliano y Sabina-Poggio Mirteto (Italia), Culemborg (Países Bajos), Elverum (Noruega), Vila Franca de Xila (Portugal), Betanzos (España), Yverdon-les-Bains, Burdorf y Bellinzona (Suiza), 1.091 varones y 1.109 mujeres, nacidos entre 1913 y 1918. El hábito tabáquico fue valorado usando un cuestionario estandarizado y categorizado en no fumadores, ex fumadores y fumadores. Se llevó a cabo una extracción de sangre y en laboratorios centrales determinaron los marcadores bioquímicos: concentraciones plasmáticas de carotenos, retinol y *-tocoferol con cromatografía líquida de alta presión (HPLC), y del ácido fólico mediante radioinmunoanálisis. El análisis estadístico de los datos se llevó a cabo con el programa R-SIGMA 2. Resultados. Los varones fumadores presentaron concentraciones (P50) más bajas de carotenos (0,34 µmol/l; p 30 µmol/l y cocientes de vitamina E/colesterol > 5,2 mmol/mol, con efectos protectores frente a enfermedades cardiovasculares según distintos estudios epidemiológicos entre los no fumadores que entre los fumadores. Conclusiones. A la vista de estos resultados la primera medida terapéutica debe ir dirigida al cese del consumo de tabaco. Los fumadores deberían consumir una dieta rica en frutas y vegetales con elevado contenido en vitaminas antioxidantes que les permitiera igualar el estado nutricional de estos micronutrientes al del grupo de los no fumadores

Introduction. Carotene and antioxidant vitamins are associated with lower arteriosclerotic risk and coronary disease. This association is supported by studies showing that smoking modifies the nutritional status of these micronutrients. Plasma levels of carotene and antioxidant vitamins in non-smokers, former smokers, and current smokers were analysed. Patients and methods. 2,200 elderly inhabitants 1,091 men, 1,109 women born between 1913 and 1918 of 19 European towns: Hamme (Belgium); Roskilde (Denmark); Chateau Renault-Amboise, Haguenau and Romans (France); Markopoulo (Greece); Anogia-Archanes (Crete); Monor (Hungary); Padua, Fara Sabina-Magliano Sabina-Poggio Mirteto (Italy); Culemborg (The Netherlands); Elverum (Norway); Vila Franca de Xila (Portugal); Betanzos (Spain); Yverdon-les-bains, Burdorf and Bellinzona (Switzerland) were randomly selected to participate in the baseline study (1988-1989) of the SENECA project (Survey in Europe on Nutrition and the Elderly, a Concerted Action) from the European Union. Smoking was assessed using a standardised questionnaire and categorised as-non, former and current smoker. Blood samples were collected and biochemical parameters: plasma levels of carotene, retinol and a-tocopherol by HPLC and folic acid by radioimmuno assay kits were measured at central laboratories, according to a strictly standardised methodology. Statistical analysis was perfomed out using the R-SIGMA 2 programme. Results. Male current smokers had levels (P50) of carotene (0.34 µmol/l; p < 0.001) and retinol (1.98 µmol/l; p < 0.01) lower than non-smokers (0.53; 2.0 µmol/l), respectively, whereas intermediate levels were observed in former smokers; carotene was the strongest parameter related to smoking consumption (r = -0.170; p < 0.006; Spearman). The marginal and deficient levels were less prevalent among non-smokers. a-tocopherol levels over 30 µmol/l or a-tocopherol/cholesterol ratios over 5.2 mmol/mol, with protective effects against cardiovascular disease, were more prevalent among non-smokers than current smokers. Conclusions. According to the results, smoking cessation would be the first measure advised. Current smokers, even former smokers, would require a diet rich in fruit and vegetables with a high antioxidant vitamin content, which would allow them to achieve the same nutritional status as non-smokers

Inhibition of cell fusion in Junin virus-infected cells by sera from Argentine hemorrhagic fever patients.

BACKGROUND: Junin virus (JV), a member of the Arenaviridae family, is the etiological agent of Argentine hemorrhagic fever (AHF). A low pH-pulse, induces fusion of Vero cells infected with JV to form syncytia, whose production can be inhibited by neutralizing antibodies against the JV major glycoprotein. OBJECTIVES: To characterize the existence of an antifusogenic activity present in sera obtained from natural infections of AHF over a 20-year period and to study both the fusogenic activity of one pathogenic and two attenuated strains of JV in Vero cells, at different pH. The study sample consisted of sera obtained from two provinces in the Argentine Republic. Vero cells grown in monolayers, were infected with different strains of JV and a 2 h pulse, at different pH, was performed. Syncytium production was evaluated 12 h later, after staining with Giemsa. Neutralization tests against the attenuated strain XJCl3 were carried out and the antifusogenic activity of immunosera was studied by incubating serum with JV-infected Vero cells. Also the fusion activity in Vero cells infected with three JV strains was assayed. RESULTS AND CONCLUSIONS: A pathogenic strain XJ exhibited the highest fusogenic activity at pH 5. Syncytium formation was prevented by patients' sera obtained from different geographical locations, independently of time of infection. However, when Vero cells were infected with XJ, a significant reduction of syncytium production was observed, though the level of inhibition was lower than that detected in other JV strains-infected cells. These results could be explained by the existence of a conserved domain on JV proteins and also antigenic heterogeneity among strains.

[Multicenter cohort of patients with HIV infection in the Madrid south-eastern metropolitan crown (COMESEM): basis, organization and initial results]. / Cohorte multicéntrica de pacientes con infección VIH de la corona metropolitana sudeste de Madrid (COMESEM): fundamentos, organización y resultados iniciales.

BACKGROUND: In order to evaluate changes related to the effect of new anti-retroviral agents and preventive programs, cohort studies of patients with HIV in our environment are needed. METHODS: Cohort study of patients diagnosed of HIV infection in the five hospitals included in the Madrid metropolitan south-eastern crown (COMESEM), which attend a population of 1,300,000 inhabitants. RESULTS: A total of 5,532 patients had been recruited until August 2001 (with a follow-up of 34,227 patients-year). The male/female ratio was 3/1. As for the transmission mechanism, 72.9% were parenteral drug users (PDU), 13.7% heterosexuals (HTX) and 8% of males having sex with males (MSM). The maximal figure of diagnosis per year corresponded to 1991 for PDU, 1993 for MSM and 1995 for heterosexuals. A decline in the number of diagnosed patients was observed for all groups, but MSM showed an increase in the year 2001 for the first time. Notably, among HTX, a growing and sustained trend was observed of patients not born in Spain (0% in 1993 and 50% in 2001; p < 0.001). CONCLUSIONS: Establishing a large cohort of HIV infected patients based upon medium-sized hospitals is possible. The analysis of data derived from this cohort allows the early detection of changes in the clinical and epidemiological profile of HIV infection.

Cohorte multicéntrica de pacientes con infección VIH de la corona metropolitana sudeste de Madrid(COMESEM): fundamentos, organización y resultados iniciales / Multicenter cohrt of patients with HIV infection in the Madrid south-eastern metropolitan crown (COMESEN): basis organization and initial results

Fundamento. Se necesitan estudios de cohorte de pacientes con infección del virus de la inmunodeficiencia humana (VIH) en nuestro medio para evaluar los cambios relacionados con el efecto de los nuevos antirretrovíricos y los programas de prevención. Métodos. Estudio de cohorte de todos los pacientes diagnosticados de infección VIH en los 5 hospitales de la corona metropolitana sudeste de Madrid (COMESEM) que atienden a una población de 1.300.000 habitantes. Resultados. Hasta agosto de 2001 se han reclutado 5.532 pacientes (con un seguimiento de 34.227 pacientes año). Hay 3 varones por cada mujer. Por vía de transmisión, el 72,9 por ciento eran usuarios de drogas por vía parenteral (UDVP), el 13,7 por ciento heterosexuales (HTX) y el 8 por ciento hombres que mantienen relaciones sexuales con hombres (HSH). El máximo de diagnósticos por año correspondió a 1991 para los UDVP, 1993 para los HSH y 1995 para los heterosexuales. En todos los grupos se ha observado un declive en el número de pacientes diagnosticados, pero los HSH en el año 2001 muestran un incremento por vez primera. Entre los HTX destaca la tendencia creciente y mantenida de pacientes no nacidos en España (0 por ciento en 1993 y 50 por ciento en 2001; p < 0,001).Conclusiones. Es posible establecer una cohorte amplia de pacientes con infección VIH basándose en hospitales de tamaño medio. El análisis de los datos derivados de esta cohorte permite objetivar cambios en el perfil clínico-epidemiológico de la infección VIH de manera precoz (AU)

Desarrollo y utilización de la historia clínica en soporte electrónico: experiencia de un servicio de nefrología de nueva creación / Electronic medical records in a recently created nephrology service

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Assessing clinical probability of organic disease in patients with involuntary weight loss: a simple score.

BACKGROUND: Involuntary weight loss (IWL) is a frequent complaint with a difficult diagnosis. Any one of a number of different diseases may be the source of the symptom. However, there is no universal clinical protocol that can help physicians study this complex syndrome. METHODS: In March 1998, we defined a diagnostic protocol for the study of IWL. IWL was defined as an involuntary and documented weight loss of at least 5% of the usual body weight in the previous 3 months. We analyzed 78 consecutive patients with IWL who came to our clinic between March 1998 and December 2000. RESULTS: An organic disease was found in 56% of cases; cancer, metabolic and digestive diseases were the most common entities. Psychiatric problems were found in 33% of cases. After extensive study, an idiopathic group of 11% was identified. The variables that were independently predictive of a final diagnosis of organic disease were: age>50 years (OR: 8.6, CI 95%: 1.7-43.6), psychiatric symptoms (OR: 0.2, CI 95%: 0.1-0.8), smoking (OR: 14.3, CI 95% 2.3-74), the presence of guide symptoms (OR: 8.0, CI 95%: 1.8-34.4), and anemia (OR: 3.1, CI 95%: 2.5-387). Sixteen percent of the patients died, more often those suffering from organic diseases. Based on multivariate regression coefficients, a clinical risk score was established. CONCLUSIONS: IWL is a complex and frequent syndrome with a 16% rate of mortality during the first year. A protocol based on clinical data can help in the management of IWL. Our clinical prediction rule may help physicians to identify those patients with IWL who are likely to have an underlying organic disease.

Involvement of vacuolar proton ATPase in Junin virus multiplication.

The role of vacuolar-proton ATPase (V-H+ ATPAse) on Junin virus (JV) replication was evaluated by analyzing the effect of specific inhibitors of the enzyme activity on different steps of virus multiplication cycle. The presence of the macrolide antibiotics bafilomycin A1 and concanamycin A during the first two hours of infection caused a significant reduction of extracellular infectious virus production and viral protein expression in Vero and BHK-21 cells. The inhibitory action of the compounds was mainly exerted at an early stage of the JV multiplication cycle, without affecting virus attachment to the cell but preventing virus penetration. A correlation between the inhibitory action of the compounds on intracellular compartments acidification and the reduction of JV yield was observed. The addition of concanamycin A at different times after infection indicated that the compound also interferes with the release of infectious particles to the extracellular medium. Although, intracellular transport of JV glycoproteins to the cell membrane, seems not to be affected as revealed by immunofluorescence staining. The results confirm that JV enters into the cell through the endocytic pathway as previously suggested by using lysosomotropic compounds.

Grave síndrome de Guillain-Barré tras primoinfección por varicela en un adulto joven / Severe Gillain Barré syndrome following initial infection with varicalla in a young adult

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Analysis of viral glycoproteins by glycosidic digestion inside a polyacrylamide gel.

We adapted the method described by Cleveland et al. (1977); (Peptide mapping by limited proteolysis in sodium dodecyl sulphate and analysis by gel electrophoresis. J. Biol. Chem. 252, 1102-1106) to study the glycosidic residues linked to the viral glycoproteins of two enveloped viruses: Junin virus (JV) and rubella virus (RV). Radioiodinated glycoproteins were obtained from purified virions, isolated from SDS-polyacrylamide gels and then hydrolysed by specific glycosidases inside a second gel. N-linked oligosaccharides, mannose and galactose were found as terminal residues in the JV-G1 glycoprotein. Mannose and N-glycans of complex hybrid type were present on RV glycoproteins.

Effects of reducing, oxidizing and alkylating agents on early steps of Junin virus multiplication.

The action of reducing, oxidizing and thiol-alkylating agents on early steps of Junin virus (JV) multiplication in Vero cells was investigated. The presence of reducing agents during virus adsorption as well as incubation of viral particles with these compounds before infection enhanced JV infectivity. On the contrary, the thiol-alkylating agent 5,5' dithiobis (2-nitrobenzoic acid) and the oxidizing compound potassium periodate showed an inhibitory effect, suggesting that sulfhydryl groups, and certain sugar moieties of viral glycoproteins play an important role in the first steps of JV infection. Also enzymatic treatment of cell monolayers and addition of concanavalin A to cultures prior to infection suggest that cellular glycoproteins are involved in virus attachment.