Long-term prognosis of HIV-infected patients with Kaposi sarcoma treated with pegylated liposomal doxorubicin.

INTRODUCTION: Incidence of Kaposi sarcoma (KS) in human immunodeficiency virus (HIV)-infected persons has dramatically decreased in the highly active antiretroviral therapy era. However, this tumor still represents the most common cancer in this population. OBJECTIVES: The objectives of this study were to evaluate long-term prognosis of HIV-infected patients with KS who had received pegylated liposomal doxorubicin (PLD) and, more specifically, to assess tumor relapse rate, mortality, and cause of death in these subjects. DESIGN: This study was a retrospective review of all patients with KS who had received PLD in centers belonging to the Caelyx/KS Spanish Group. Kaplan-Meier analysis and univariate and multivariate Cox-regression analysis were used to assess the rate of and factors associated with relapse and death through January 2006. RESULTS: A total of 98 patients received PLD from September 1997 through June 2002. Median follow-up after initiation of treatment was 28.7 months (interquartile range, 6.6-73.2 months); during follow-up, 29 patients died (a mortality rate of 14.6% per year). In 9 patients (31%), the cause of death was related to the appearance of other tumors (including 7 lymphomas, 1 gastrointestinal adenocarcinoma, and 1 tongue epidermoid cancer). Death caused by progression of KS occurred in 3 cases. Death risk was inversely related to CD4(+) cell counts at the end of follow-up (hazard ratio for every increase in CD4(+) cell count of 100 cells/microL, 0.7; 95% confidence interval, 0.5-0.9). A relapse study was performed for 61 patients who had complete or partial response to PLD and who attended a control visit after treatment completion. After a median follow-up of 50 months (interquartile range, 17.2-76 months), 8 patients (13%) had experienced relapse; 5 of these patient experienced relapse within the first year after stopping PLD. The only factor that was independently related to risk of relapse was having a CD4(+) cell count >200 cells/microL at baseline (hazard ratio, 6.2; 95% confidence interval, 1.2-30). Lower CD4(+) cell count at the end of follow-up was marginally associated with relapse (hazard ratio for every increase in CD4(+) cell count of 100 cells/microL, 0.7; 95% confidence interval, 0.6-1.01). CONCLUSIONS: Treatment of KS with PLD in HIV-infected patients is followed by a low relapse rate, with most relapses occurring during the first year after stopping chemotherapy. However, the mortality rate in this population was high, in part because of an unexpectedly high incidence of other tumors, mainly lymphomas.

Evolution of bone mineral density in AIDS patients on treatment with zidovudine/lamivudine plus abacavir or lopinavir/ritonavir.

OBJECTIVES: The aim of the study was to determine the factors that may contribute to decreases in bone mineral density (BMD) in patients with AIDS. METHODS: This was a prospective, non-randomized study. Dual X-ray absorptiometry (DXA) was used to determine the BMD of the lumbar spine, femoral neck and distal radius in treatment-naïve HIV-infected male patients with AIDS before and after 1 year of treatment with zidovudine (ZDV)/lamivudine (3TC) plus abacavir (ABC) or lopinavir/ritonavir (LPV/r). RESULTS: Basal DXA was performed in 50 patients with CD4 counts <200 cells/microL and/or any AIDS-defining condition. Thirty-two patients completed 1 year with full adherence (17 on ABC and 15 on LPV/r) and a second DXA was then performed. At baseline, 19% had osteopenia at the lumbar spine and 19% at the femoral neck. Low body weight was related to low BMD. After 48 weeks, BMD loss was significant at the three locations. The percentage of BMD loss at the femoral neck tended to be greater in the lopinavir group (5.3 vs. 3.2%, P=0.058). The differences became significant at the lumbar spine (5.7 vs. 2.7%, P=0.044). In the multivariate analysis, the treatment with LPV/r remained associated with bone loss at the lumbar spine. CONCLUSIONS: Osteopenia is frequent in treatment-naïve HIV-infected men with AIDS. Bone loss is higher with LPV/r-based regimens compared with triple nucleoside reverse transcriptase inhibitors.

First epidemic of echovirus 16 meningitis in Cuba.

From April to September 2000, an epidemic of aseptic meningitis spread throughout Cuba, with 16,943 reported cases. Virologic studies identified echovirus 16 as the cause of this epidemic. This is the first reported isolate of echovirus 16 from patients with viral meningitis in Cuba.

[Hemorrhagic pneumonia caused by influenza virus: virological diagnosis]. / Neumonía hemorrágica por Influenza virus: diagnóstico virológico.

The paper presents the case of a female patient who was admitted to "Calixto García" General Hospital with respiratory distress and hypovolemic or septic shock. She was diagnosed with viral hemorrhagic pneumonia. From the endotracheal secretion taken as a sample, the influenza virus was isolated as etiological agent, which, through the hemaglutination inhibition technique, was characterized as a strain belonging to H3N2 subtype, very similar to strain A/Johannesburg/33/94 from the antigenic viewpoint. The patient recovered satisfactorily after being treated with rivabirin.

Isolation and identification of adenovirus in hospitalized children, under five years, with acute respiratory disease, in Havana, Cuba.

Nine Adenovirus (Ad) strains isolated in Cuba, from 128 nasopharingeal swab specimens of children below five years old, with acute respiratory diseases, during 1996 and 1997, were studied by restriction enzyme analysis of genomic DNA with two endonucleases BamH I and Sma I. All different fragment patterns were compared with the respective prototypes. The identified adenoviruses were Ad 1 (n=4), Ad 2 (n=1) and Ad 6 (n=4). Males were more frequently infected than females. The analysis of the occurrence of these Adenovirus strains of subgenus C revealed that Ad 1 and Ad 6 were the predominant serotypes in 1996 and in 1997, respectively.

[Rapid diagnosis of principal respiratory viruses in the city of Havana, 1995-97]. / Diagnóstico rápido de los principales virus respiratorios en Ciudad de la Habana, 1995-97.

Acute respiratory diseases (ARD) are the most common infections in humans and difficult to prevent. Viruses have been recognized as predominant ethiological agents. In Cuba, ARD constitute a major problem of health and are the first cause of morbidity and important cause of mortality. In this paper, rapid diagnosis was performed to 516 clinical samples which arrived to the Reference Respiratory Viruses Laboratory of the Pedro Kourí Institute of Tropical Medicine (IPK) from different parts of Havana City during 1995, 1996 and 1997. The results obtained have shown 218 positive samples (Influenza A, 89; respiratory syncytial virus 52; Influenza B, 45; Adenovirus, 13; human parainfluenza virus(HPIV)-1, 6; HPIV-2, 3 and HPIV-3, 10). Influenza A was the virus most frequently found in adults, whereas in closed population of teen-agers and adults, Influenza B was frequently found. Furthermore, respiratory syncytial virus was the most important pathogen in children's under 1 year of age.

Detection of respiratory syncytial virus in nasopharyngeal secretions by 24-well plate precentrifugation assay using a monoclonal antibody against F protein.

BACKGROUND: Respiratory syncytial virus (RSV) is responsible for 50% of all bronchiolitis and 25% of pneumonia cases during the first month of life. Detection of the RSV antigen by immunofluorescence in exfoliated nasal epithelium or by other methods in nasopharyngeal swabs is useful in the potentially infected patient because results are available within a few hours. In contrast, RSV antigen detection in cell culture may require as much as 3 weeks. METHODS: Three methods for detection of respiratory syncytial virus in 131 clinical respiratory specimens from patients with acute respiratory disease and bronchiolitis were compared utilizing the following: a precentrifugation immunofluorescence assay using Hep-2 cells, indirect immunofluorescence assay, and conventional tube cell culture using Hep-2 cells. RESULTS: Respiratory syncytial virus was identified in 36 specimens by the three methods previously described. The virus was recovered in 41 (31.3%) samples by precentrifugation immunofluorescence assay, 40 (30.5%) were identified by the immunofluorescence technique, and 38 (29.0%) cases were positive by conventional cell culture. The sensitivity of the precentrifugation assay in relation to the immunofluorescence technique was 90%, the specificity 94.5%, and the agreement, 96.2%. A positive predictive value of 90.2% was obtained. Sensitivity, specificity, agreement, and positive predictive values obtained by the precentrifugation assay variant compared to the conventional cell were 90.8%, 94.5%, 93.1%, and 87.8%, respectively. CONCLUSIONS: The precentrifugation immunofluorescence assay method was as sensitive as the remainder of the methods used in our study and represents a valid alternative for rapid detection of respiratory syncytial virus in clinical samples.

Diagnóstico rápido de los principales virus respiratorios en Ciudad de la Habana, 1995-97 / Rapid diagnosis of principal respiratory viruses in the city of Havana, 1995-97

Acute respiratory diseases (ARD) are the most common infections in humans and difficult to prevent. Viruses have been recognized as predominant ethiological agents. In Cuba, ARD constitute a major problem of health and are the first cause of morbidity and important cause of mortality. In this paper, rapid diagnosis was performed to 516 clinical samples which arrived to the Reference Respiratory Viruses Laboratory of the Pedro KourÝ Institute of Tropical Medicine (IPK) from different parts of Havana City during 1995, 1996 and 1997. The results obtained have shown 218 positive samples (Influenza A, 89; respiratory syncytial virus 52; Influenza B, 45; Adenovirus, 13; human parainfluenza virus(HPIV)-1, 6; HPIV-2, 3 and HPIV-3, 10). Influenza A was the virus most frequently found in adults, whereas in closed population of teen-agers and adults, Influenza B was frequently found. Furthermore, respiratory syncytial virus was the most important pathogen in children's under 1 year of age.

Diagnóstico rápido de los principales virus respiratorios en Ciudad de la Habana, 1995-97 / Rapid diagnosis of principal respiratory viruses in the city of Havana, 1995-97

Acute respiratory diseases (ARD) are the most common infections in humans and difficult to prevent. Viruses have been recognized as predominant ethiological agents. In Cuba, ARD constitute a major problem of health and are the first cause of morbidity and important cause of mortality. In this paper, rapid diagnosis was performed to 516 clinical samples which arrived to the Reference Respiratory Viruses Laboratory of the Pedro KourY Institute of Tropical Medicine (IPK) from different parts of Havana City during 1995, 1996 and 1997. The results obtained have shown 218 positive samples (Influenza A, 89; respiratory syncytial virus 52; Influenza B, 45; Adenovirus, 13; human parainfluenza virus(HPIV)-1, 6; HPIV-2, 3 and HPIV-3, 10). Influenza A was the virus most frequently found in adults, whereas in closed population of teen-agers and adults, Influenza B was frequently found. Furthermore, respiratory syncytial virus was the most important pathogen in childrens under 1 year of age.(AU)

Diagnóstico rápido de los principales virus respiratorios en Ciudad de la Habana, 1995-97. / [Rapid diagnosis of principal respiratory viruses in the city of Havana, 1995-97]

Acute respiratory diseases (ARD) are the most common infections in humans and difficult to prevent. Viruses have been recognized as predominant ethiological agents. In Cuba, ARD constitute a major problem of health and are the first cause of morbidity and important cause of mortality. In this paper, rapid diagnosis was performed to 516 clinical samples which arrived to the Reference Respiratory Viruses Laboratory of the Pedro Kourí Institute of Tropical Medicine (IPK) from different parts of Havana City during 1995, 1996 and 1997. The results obtained have shown 218 positive samples (Influenza A, 89; respiratory syncytial virus 52; Influenza B, 45; Adenovirus, 13; human parainfluenza virus(HPIV)-1, 6; HPIV-2, 3 and HPIV-3, 10). Influenza A was the virus most frequently found in adults, whereas in closed population of teen-agers and adults, Influenza B was frequently found. Furthermore, respiratory syncytial virus was the most important pathogen in childrens under 1 year of age.

Antigenic and genetic characterization of twenty-six strains of human respiratory syncytial virus (subgroup A) isolated during three consecutive outbreaks in Havana city, Cuba.

Twenty-six human respiratory syncytial virus strains (subgroup A) isolated from three outbreaks in Havana City during the period 1994/95, 1995/96 and 1996/97 were analyzed to determine their antigenic and genetic relationships. Analyses were performed by monoclonal antibodies and restriction mapping (N gene) following amplification of the select region of the virus genome by polymerase chain reaction. All isolated strains were classified as subgroup A by monoclonal antibodies and they showed a restriction pattern NP4 that belonged to subgroup A. Thus the results obtained in this work, showed a close relation (100%) between antigenic and genetic characterization of the isolated strains in our laboratory. These methods permit the examination of large numbers of isolates by molecular techniques, simplifying the researchs into the molecular epidemiology of the virus.

Unusual antigenic and genetic characteristics of human respiratory syncytial viruses isolated in Cuba.

The G protein of 23 strains of human respiratory syncytial virus isolated in Havana, Cuba, between October 1994 and January 1995 was analyzed at the antigenic and genetic level. All viruses reacted with 10 of 11 antibodies specific for the Long strain. Moreover, the G protein gene of the Cuban isolates had only five nucleotide differences from the sequence of the Long gene. The homogeneity of the Cuban isolates and their resemblance to an ancient strain, such as Long, are at odds with previous findings for viruses isolated in countries with a temperate climate and different socioeconomic status. The G proteins of three of four other viruses isolated in Havana 2 years later (1996) were also identical to those of the 1994-to-1995 isolates, and the fourth virus had a single extra nucleotide difference. This, again, is unusual, since no identical viruses had been isolated in different epidemics previously. The singular characteristics of the Cuban isolates reported here are discussed in terms of the epidemiological, climatic, and socioeconomic characteristics of Cuba.

[The incidence of adenoviruses in viral conjunctivitis]. / Incidencia de los Adenovirus en las conjuntivitis virales.

A study about the incidence of Adenovirus on viral conjunctivitis was conducted. A sample design was made and samples of conjunctival exudate were taken from 150 patients with diagnosis of apparently viral conjunctivitis at the "Ramón Pando Ferrer" Ophthalmological Hospital from July to December, 1994. Samples were inoculated in cell culture and the indirect immunofluorescence technique was applied to those with a cytopathogenic effect that suggested infection due to Adenovirus. Monoclonal antibodies were used against Adenovirus allowing to identify them as part of the Adenoviridae family. The hemagglutination technique was used with erythrocytes of monkeys and rats as an indicator system in order to group the isolates previously identified by the indirect immunofluorescence technique. Later on, it was made an analysis by restriction enzymes of the viral genome to enable typing. The results of this study showed an incidence of Adenovirus on viral conjunctivitis of 20%, with a confidence interval between 14 and 26% and a reliability index of 95%. It was proved that serotype 37 caused conjunctivitis more frequently.

Molecular characterization of an outbreak of respiratory syncytial virus (subgroup A) in Havana, Cuba, by monoclonal antibodies and restriction mapping (N gene).

Twenty-one respiratory syncytial virus (RSV) strains isolated from one outbreak in Havana, Cuba (1994 to 1995), were analyzed to determine their relatedness. All isolated strains were classified as subgroup A by monoclonal antibodies. Of 21 RSV strains examined, 20 were classified as having restriction pattern NP4 and only 1 was classified as having restriction pattern NP5.

Analysis of respiratory syncytial virus in clinical samples by reverse transcriptase-polymerase chain reaction restriction mapping.

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.

[Application of the polymerase chain reaction for detecting respiratory syncytial virus]. / Aplicación de la reacción en cadena de la polimerasa para la identificación del virus sincitial respiratorio.

The polymerase chain reaction (PCR) was developed in order to identify the respiratory syncytial virus by using the reference strain. The high sensitivity and specificity obtained show the PCR utility for detecting the RSV genoma and its application on the diagnosis.

[Evaluation of a mouse anti-IgG-fluorescein conjugate using indirect immunofluorescence and flow cytometry techniques]. / Evaluación de un conjugado anti-IgG de ratón-fluoresceína mediante técnicas de inmunofluorescencia indirecta y citometría de flujo.

An immunoglobulin G of mouse was purified from sera by affinity chromatography in protein A. The rabbits whose sera were able to recognize the antigen injected by double immunodiffusion were immunized with this preparation. The antibodies were precipitated from the rabbit's serum and purified by ion exchange chromatography. This preparation was conjugated to fluorescin isothiocyanate according to the conventional technique. The conjugated obtained was evaluated with the reference strains of Parainfluenza virus 1, 2, 3; Adenovirus; respiratory syncytial virus; and influenza virus A and B, by an indirect immunofluorescence technique and HIV positive samples by flow citometry. Specific monoclonal antibodies were used in both cases. Clinical specimens of patients with acute respiratory infection were evaluated.

[Estimation of IgG Adenovirus antibody titers using a standard curve and an indirect ultramicroELISA essay]. / Estimación del título de anticuerpos IgG a Adenovirus utilizando una curva patrón y un ensayo de ultramicroELISA indirecto.

It was possible to standardize a procedure which combined an indirect microELISA assay with a standard curve and that allowed to estimate the titre of IgG antibodies to Adenovirus in samples of human serum, using only one dilution of these. Based on the end-point titre previously determined for a panel of 117 serum samples, we selected 90 of these samples (r2 = 0.98) to build 4 standard curves that related the natural logarithm of the fluorescence responses to the natural logarithm of the end-point titre for a wide range of serum dilutions (1:40 = 1:320). It was selected the curve corresponding to serum dilution 1:40 (r2 = 0.81), which made possible an optimum utilization of those accessories designed to handle the volumes in the ultramicro range and, therefore, the automation of the whole procedure. The results obtained as regards the complement fixation test (100% of sensitivity and 97.3% of specificity) support the use of this method in our laboratory as a complementary tool to carry out seroepidemiological studies on a large scale and with diagnostic ends.

[Subgroups classification of strains of the respiratory syncytial virus isolated in an outbreak in Ciudad de La Havana]. / Clasificación en subgrupos de cepas del virus sincitial respiratorio aisladas de un brote en Ciudad de La Habana.

A high number of acute respiratory diseases was detected among children under one year admitted in a hospital of Havana City. 25 respiratory syncytial virus strains were obtained from 93 patients studied. Viral isolations were multiplied in HEP-2 cells and after observing a cytopathic effect of 80%, they were classified into subgroups by the indirect immunofluorescence technique, using anti-protein G antibodies from the respiratory syncytial virus. All the samples studied were classified within subgroup A. It is the first time a study like this is conducted in our country, which allowed us to deepen into the viral cause of these diseases and to know that the subgroup A of the respiratory syncytial virus circulated during the outbreak.