Extracellular acidosis enhances Zika virus infection both in human cells and ex-vivo tissue cultures from female reproductive tract.

Zika virus (ZIKV) is a flavivirus transmitted by mosquitoes of the genus Aedes, but unlike other flaviviruses, ZIKV can be sexually transmitted by vaginal intercourse. The healthy vaginal pH ranges from 4.0 to 6.0, reaching values of 6.0-7.0 after semen deposition. Here, we report that low extracellular pH values (range 6.2-6.6) dramatically increase ZIKV infection on cell lines of different origin including some derived from the female genital tract and monocyte-derived macrophages. Furthermore, low pH significantly increased ZIKV infection of human ectocervix and endocervix cultured ex-vivo. Enhancement of infection by low pH was also observed using different ZIKV strains and distinct methods to evaluate viral infection, i.e. plaque assays, RT-PCR, flow cytometry, and fluorescence microscopy. Analysis of the mechanisms involved revealed that the enhancement of ZIKV infection induced by low pH was associated with increased binding of the viral particles to the heparan sulphate expressed on the target cell surface. Acidosis represents a critical but generally overlooked feature of the female genital tract, with major implications for sexual transmission diseases. Our results suggest that low vaginal pH might promote male-to-female transmission of ZIKV infection.

A pilot study of a stepped-care brief intervention to help psychologically-distressed women displaced by conflict in Bogotá, Colombia.

BACKGROUND: Colombia's 6.5 million internally displaced persons (IDPs) have been exposed to trauma, loss, and hardships. Common mental disorders (CMDs) are prevalent in this group, yet there are few evidence-based psychosocial interventions for this population. We assessed the feasibility and acceptability of a stepped-care intervention for women IDPs in Bogota, Colombia. METHODS: Feasibility to recruit participants for an intervention trial, to screen for CMDs and displacement-related traumas, to refer high-risk cases to professional consultation, to implement evidence-based interpersonal counseling (IPC) for women with diagnosed CMDs, to retain participants in the intervention, and to conduct follow-up assessments was assessed. Assessment instruments were validated. The intervention was delivered by trained outreach personnel. Intervention acceptability was assessed by monitoring session attendance, dropout rates, and satisfaction. Potential efficacy was evaluated with pre- and post-intervention measures of CMDs. RESULTS: We recruited 279 women IDPs into the intervention. On screening, 177 (63.4%) had symptom levels suggesting a CMD. Participants endorsed a wide range of displacement-related exposures. Most participants receiving IPC decreased their symptom levels at follow-up. Many participants did not complete the recommended number of IPC sessions; loss to follow-up was 30%. The performance of the outreach personnel improved after the initial intervention team was replaced with community members trained to deliver the intervention. The Bogotá health system was unable to reliably accommodate emergency psychiatric referrals. CONCLUSIONS: The IPC intervention shows promise, but significant challenges remain for improving reach, adherence, and participant retention. We identified strategies and partnerships to redress some of the main study limitations.

Seminal vesicle fluid increases the efficacy of intravaginal HSV-2 vaccination.

Once considered merely as a vehicle for spermatozoa, it is now clear that seminal plasma (SP) induces a variety of biological actions on the female reproductive tissues able to modulate the immune response against paternal antigens. To our knowledge, the influence of SP on the immune response against sexually transmitted pathogens has not been yet evaluated. We here analyzed whether the seminal vesicle fluid (SVF), which contributes almost 60% of the SP volume in mice, could modulate the immune response against herpes simplex virus type 2 (HSV-2). We found that SVF does not modify the course of primary infection, but markedly improved protection conferred by vaginal vaccination with inactivated HSV-2 against a lethal challenge. This protective effect was shown to be associated to a robust memory immune response mediated by CD4+ and CD8+ T cells in both the lymph nodes draining the vagina and the vaginal mucosa, the site of viral replication. In contrast with the widespread notion that SP acts as an immunosuppressive agent, our results suggest that SVF might improve the female immune response against sexually transmitted pathogens.

Acetylcholine polarizes dendritic cells toward a Th2-promoting profile.

BACKGROUND: A growing body of research shows a reciprocal regulation between the neural and immune systems. Acetylcholine (ACh) is the most important parasympathetic neurotransmitter, and increasing evidence indicates that it is able to modulate the immune response. Interestingly, in recent years, it has become clear that immune cells express a non-neuronal cholinergic system, which is stimulated in the course of inflammatory processes. We have previously shown that dendritic cells (DC) express muscarinic receptors, as well as the enzymes responsible for the synthesis and degradation of ACh. Here, we analyzed whether ACh could also modulate the functional profile of DC. METHODS: Dendritic cells were obtained from monocytes cultured for 5 days with GM-CSF+IL-4 or isolated from peripheral blood (CD1c+ DC). The phenotype of DC was evaluated by flow cytometry, the production of cytokines was analyzed by ELISA or intracellular staining and flow cytometry, and the expression of muscarinic and nicotinic receptors was evaluated by flow cytometry or qRT-PCR. RESULTS: Treatment of DC with ACh stimulated the expression of the Th2-promoter OX40L, the production of the Th2-chemokines MDC (macrophage-derived chemokine/CCL22) and TARC (thymus and activation-regulated chemokine/CCL17), and the synthesis of IL-4, IL-5, and IL-13 by T cells, in the course of the mixed lymphocyte reaction (MLR). Moreover, we found that the stimulation of OX40L, HLA-DR, and CD83 expressions in DC induced by the Th2-promoting cytokine TSLP, as well as the production of IL-13, IL-4, and IL-5 by T cells in the course of the MLR, was further enhanced when DC were treated with TSLP plus ACh, instead of TSLP or ACh alone. CONCLUSIONS: Our observations suggest that ACh polarizes DC toward a Th2-promoting profile.

Evaluation of the efficacy of intramuscular versus intramammary treatment of subclinical Streptococcus agalactiae mastitis in dairy cows in Colombia.

A randomized controlled trial was performed in 17 Colombian dairy herds to determine the cure risk among cows subclinically infected with Streptococcus agalactiae exposed to 2 antibiotic therapies. Composite milk samples were collected before milking at the onset of the trial (pretreatment) and 2 subsequent times over a period of approximately 63 d. The intramammary application (IMM) of ampicillin-cloxacillin was compared with the intramuscular application (IM) of penethamate hydriodide, and cure risks after an initial and retreatment application were assessed. Cure risk after the initial treatment was higher (82.4%) for the IMM treatment than for IM therapy (65.8%). However, no difference was observed in the cure risk of refractory cases after retreatment (IMM=52.6% vs. IM=51.2%). The cumulative cure risk (both initial and retreatment) was 90.4 and 82.9% for the IMM and IM products, respectively. A 2-level random effects logistic model that controlled for pretreatment cow-level somatic cell count, indicated that IM treatment (odds ratio=0.37) had a lower cure risk than IMM and a tendency for a lower cure risk with increasing baseline somatic cell count. Our findings suggest that both products and administration routes can reduce the prevalence of S. agalactiae in affected herds, but the IMM product had a better efficacy in curing the infection. In addition to the treatment protocol, the cow somatic cell count should be considered when making management decisions for cows infected with S. agalactiae.

Fucosylated clusterin in semen promotes the uptake of stress-damaged proteins by dendritic cells via DC-SIGN.

STUDY QUESTION: Could seminal plasma clusterin play a role in the uptake of stress-damaged proteins by dendritic cells? SUMMARY ANSWER: Seminal plasma clusterin, but not serum clusterin, promotes the uptake of stress-damaged proteins by dendritic cells via DC-SIGN. WHAT IS KNOWN ALREADY: Clusterin is one of the major extracellular chaperones. It interacts with a variety of stressed proteins to prevent their aggregation, guiding them for receptor-mediated endocytosis and intracellular degradation. The concentration of clusterin in semen is almost 20-fold higher than that found in serum, raising the question about the role of seminal plasma clusterin in reproduction. No previous studies have analyzed whether seminal plasma clusterin has chaperone activity. We have previously shown that seminal plasma clusterin, but not serum clusterin, expresses an extreme abundance of fucosylated glycans. These motifs enable seminal plasma clusterin to bind DC-SIGN with very high affinity. STUDY DESIGN, SIZE, DURATION: In vitro experiments were performed to evaluate the ability of seminal plasma clusterin to inhibit the precipitation of stressed proteins, promoting their uptake by dendritic cells via DC-SIGN (a C-type lectin receptor selectively expressed on dendritic cells (DC)). Moreover, the ability of seminal plasma clusterin to modulate the phenotype and function of DCs was also assessed. PARTICIPANTS/MATERIALS, SETTING, METHODS: Clusterin was purified from human semen and human serum. Catalase, bovine serum albumin, glutathione S-transferase, and normal human serum were stressed and the ability of seminal plasma clusterin to prevent the precipitation of these proteins, guiding them to DC-SIGN expressed by DCs, was evaluated using a fluorescence-activated cell sorter (FACS). Endocytosis of stressed proteins was analyzed by confocal microscopy and the ability of seminal plasma clusterin-treated DCs to stimulate the proliferation of CD25+FOXP3+CD4+ T cells was also evaluated by FACS. MAIN RESULTS AND THE ROLE OF CHANCE: Seminal plasma clusterin interacts with stressed proteins, inhibits their aggregation (P < 0.01) and efficiently targets them to dendritic cells via DC-SIGN (P < 0.01). DCs efficiently endocytosed clusterin-client complexes and sorted them to degradative compartments involved in antigen processing and presentation. Moreover, we also found that the interaction of seminal plasma clusterin with DC-SIGN did not change the phenotype of DCs, but stimulates their ability to induce the expansion of CD25+FOXP3+CD4+ T lymphocytes (P < 0.05 versus control). LIMITATIONS, REASONS FOR CAUTION: All the experiments were performed in vitro; hence the relevance of our observations should be validated in vivo. WIDER IMPLICATIONS OF THE FINDINGS: Our results suggest that by inducing the endocytosis of stress-damaged proteins by DCs via DC-SIGN, seminal plasma clusterin might promote a tolerogenic response to male antigens, thereby contributing to female tolerance to seminal antigens. STUDY FUNDING/COMPETING INTERESTS: The present research was supported by the Consejo Nacional de Investigaciones Científicas y Técnicas, the Buenos Aires University School of Medicine, and the Agencia Nacional de Promoción Científica y Tecnológica (Argentina). The authors have no conflicts of interest to declare.

Prostaglandins in semen compromise the immune response against sexually transmitted pathogens.

Seminal plasma is not just a spermatozoa carrier. It induces the expression of inflammatory cytokines and chemokines and a massive infiltration of neutrophils, monocytes and dendritic cells in the female genital mucosa after coitus, enabling the innate immune system to fight against sexually transmitted pathogens. However, exposure to seminal plasma not only turns on an inflammatory response but also induces regulatory mechanisms that allow the fetus (a semiallograft) to grow and develop in the uterus. In mouse models it has been shown that seminal plasma induces the expansion of regulatory T cells specific to seminal Ags in the receptive partner, thus promoting tolerance to paternal alloantigens and avoiding allogeneic fetal rejection. These mechanisms appear to be mainly induced by prostaglandins of the E series (PGE) and TGF-ß, which are present at huge concentrations in the seminal plasma. Moreover, we have recently shown that exposure to seminal plasma induces the differentiation of dendritic cells into a tolerogenic profile through a mechanism dependent on the activation of the prostanoid receptors EP2 and EP4 by seminal PGE. Our hypothesis proposes that this tolerogenic response induced by seminal PGE, while promoting fertility by inducing tolerance toward paternal alloantigens, might also compromise the development of the adaptive immune response against sexually transmitted pathogens in the receptive partner.

Barium selenate supplementation and its effect on intramammary infection in pasture-based dairy cows.

A significant proportion of cattle receive inadequate dietary Se because of its low content in soils and pastures of various regions of the world. Several economically important diseases in dairy cows, such as mastitis, have been associated with Se deficiency. The objective of this study was to evaluate the effect of a single injection of a long-acting form of Se at drying off on the risk and incidence rate of new intramammary infections and on milk somatic cell count in the subsequent lactation in pasture-based dairy cows. Forty-nine Chilean Holstein-Friesian cows were fed a diet containing <0.05 mg of Se/kg of ration dry matter. During the dry period, cows were allocated to 1 of 2 groups, a supplemented (n=24) group treated with a single subcutaneous injection of barium selenate 2 mo before calving and a control group (n=25) that remained unsupplemented. Duplicate foremilk samples were aseptically collected within 6 d after calving and every 2 wk until drying-off for bacteriological culture. Milk samples were also collected monthly for somatic cell count evaluation. Blood samples were collected before treatment and at 30, 90, 180, and 270 d after treatment for analysis of blood glutathione peroxidase (GPx) activity. The activity of glutathione peroxidase was higher in supplemented cows 30 d after the injection until the end of the study. The risk and incidence rate of new intramammary infections was not affected by supplementation. A progressive increase in somatic cell count was observed throughout lactation, but there was no effect of supplementation. In conclusion, a one-time injection of barium selenate 2 mo before calving in these pasture-based dairy cows did not affect udder health in the subsequent lactation, indicating that Se basal intake was adequate for preventing subclinical mastitis in pasture-based cows in southern Chile.

Viral reactivation and pseudotype production in an in vitro superinfection system with two different strains of HIV-1.

Viral production and variability of HIV-1 is normally high in vivo causing the necessary conditions for cellular superinfection. In order to evaluate the superinfection dynamics in vitro, H9HTLVIIIB cell line was superinfected with HIVMN. Superinfected cells showed nearly 50% cell mortality at day 1 post-superinfection (ps), which increased significantly up to day 4 ps. Superinfecting genome was detectable until day 10 ps. The superinfecting strain was found in the supernatant only on day 1 ps, but was recovered up to day 4 ps by coculture with non-infected cells. The existing strain (HIVHXB2) was recovered throughout the studied period. Pseudotype formation by the HIVHXB2 genome and envelope proteins of the superinfecting strain (HIVMN) was observed from day 1 to 6 ps. Viral production was increased by 1.7 LOG in superinfected cells from day 1 ps. Both viral production increase and pseudotype formation could be relevant for HIV pathogenesis in vivo.

Influence of formulation and process variables on in vitro release of theophylline from directly-compressed Eudragit matrix tablets.

Extended-release theophylline (TP) matrix tablets were prepared by direct compression of drug and different pH-dependent (Eudragit L100, S100 and L100-55) and pH-independent (Eudragit RLPO and RSPO) polymer combinations. The influence of varying the polymer/polymer (w/w) ratio and the drug incorporation method (simple blend or solid dispersion) was also evaluated. Drug release, monitored using the Through Flow Cell system, markedly depended on both the kind of Eudragit polymer combinations used and their relative content in the matrix. Maintaining a constant 1:1 (w/w) drug/polymers ratio, the selection of appropriate mixtures of pH-dependent and pH-independent polymers enabled achievement of a suitable control of TP release. In particular, matrices with a 0.7:0.3 w/w mixture of Eudragit L100-Eudragit RLPO showed highly reproducible drug release profiles, with an almost zero-order kinetic, and allowed 100% released drug after 360 min. As for the effect of the drug incorporation method, simple blending was better than the solid dispersion technique, which not only did not improve the release data reproducibility, but also caused, unexpectedly, a marked slowing down in drug release rate.

The effect of D-003 (10 mg/day) on biochemical parameters of bone remodelling in postmenopausal women: a randomized, double-blind study.

Biphosphonates, which are antiresorptive agents used to treat osteoporosis, inhibit the mevalonate pathway, preventing protein prenylation and inhibiting osteoclast activity. Statins decrease cholesterol biosynthesis by blocking the mevalonate pathway and have been reported to have beneficial effects on bone. D-003 is a mixture of high molecular weight acids purified from sugarcane wax that inhibits cholesterol biosynthesis before mevalonate production. D-003 prevents bone loss and resorption in rats with osteoporosis induced with ovariectomy or corticoids. Biochemical markers of bone turnover are used to monitor the short-term efficacy of antiosteoporotic therapy. This randomized, double-blind, placebo-controlled study was undertaken to investigate the short-term effects of D-003 (10 mg/day) on biochemical markers of bone turnover in postmenopausal women with low bone mineral density (BMD). After 4 weeks on a low-fat diet, 34 women were randomized to D-003 (10 mg/day) or placebo for 6 months. Pre- and post-treatment samples were analyzed for urinary excretion of deoxypyridinoline (DPD)/creatinine (Cr), a marker of bone resorption, and serum bone specific alkaline phosphatase (BSAP), a marker of bone formation. The effects on lipid profile and safety indicators, as well as adverse events (AE), were investigated. D-003 (10 mg/day) lowered urinary excretion of tDPD/Cr versus baseline (20.6%) (p < 0.001) and placebo (33.7%) (p < 0.01), but did not modify serum BSAP. D-003 decreased low-density lipoprotein-cholesterol (LDL-C) (32.8%), total cholesterol (TC) (16.4%) and the TC/high-density lipoprotein-cholesterol (HDL-C) ratio (34.7%), increased HDL-C (30.3%) (p < 0.001) and did not modify triglycerides. The effects on these variables were significant as early as 3 months after treatment initiation. D-003 was well tolerated. Three patients (one in the placebo group and two in the D-003 group) withdrew from the study. Two of these withdrawals were due to AE: abdominal pain (placebo) and heartburn (D-003). Five patients (four in the placebo group [22.2%] and one in the D-003 group [6.3%]) reported mild AE. In conclusion, D-003 (10 mg/day) reduced urinary excretion of tDPD/Cr, a bone resorption marker and did not change serum BSAP, a bone formation marker, while it lowered cholesterol in study patients. These preliminary results suggest that D-003 could be useful in treating postmenopausal women with low BMD. However, the potential value of D-003 in treating or preventing osteoporosis deserves further clinical investigation.

Oxytocin and prostaglandins F2alpha and E2 do not enhance HIV antigen production in vitro.

Oxytocin and prostaglandins (PGs) are hormones involved in labor and are used clinically for its induction. In this study the effect of oxytocin, PGF(2alpha), and PGE(2) on Humour immunodeficiency virus-1 production in acutely and persistently infected cells was measured. No significant effect on p24 antigen production was found with oxytocin or PGs, except for a transient decrease in persistently infected cells treated with 1 micro M PGF(2alpha). These results showed that oxytocin and PGs could be used clinically for labor induction without any direct enhancement in viral production. Besides, the results with PGF(2alpha) at the highest concentration studied may indicate a pharmacological effect.

[Streptococcus pneumoniae: evolution of antibiotic resistance in a pediatric hospital in Córdoba, Argentina]. / Streptococcus pneumoniae: evolución de la resistencia a los antimicrobianos en un hospital de niños de Córdoba, Argentina.

The wide variety of prevalence of antimicrobial resistant Streptococcus pneumoniae in different countries confirms the importance of determining local patterns of resistance. From 1992 to 2000, we studied the pattern of antimicrobial resistance in S. pneumoniae and its evolution along the years, using 468 strains isolated in the Hospital de Niños de Córdoba. A total of 177 isolates (37.8%) were not susceptible to penicillin, with 19% intermediate and 18.8% resistant strains. High and intermediate resistance levels to cefotaxime were 4.9% and 10.9%, respectively. Decreased susceptibility to trimethoprim/sulfamethoxazole (TMS), erythromycin, chloramphenicol, and rifampin was found in 194 isolates (41.5%), 32 (6.8%), 13 (2.8%) and 3 (0.6%), respectively. No isolates resistant to vancomycin were detected. The most commonly combined resistance patterns were: penicillin/TMS (35.6%) and penicillin/TMS/cefotaxime (11.8%). This study highlights the increased rate of drug resistant S. pneumoniae during the last years, and the importance of antimicrobial resistance surveillance of adequate empirical therapy involving local and regional susceptibility patterns.

Streptococcus pneumoniae: evolución de la resistencia a los antimicrobianos en un hospital de niños de Córdoba, Argentina / Streptococcus pneumoniae: evolution of antibiotic resistance in a pediatric hospital in Córdoba, Argentina

The wide variety of prevalence of antimicrobial resistant Streptococcus pneumoniae in different countries confirms the importance of determining local patterns of resistance. From 1992 to 2000, we studied the pattern of antimicrobial resistance in S. pneumoniae and its evolution along the years, using 468 strains isolated in the Hospital de Niños de Córdoba. A total of 177 isolates (37.8) were not susceptible to penicillin, with 19 intermediate and 18.8 resistant strains. High and intermediate resistance levels to cefotaxime were 4.9 and 10.9, respectively. Decreased susceptibility to trimethoprim/sulfamethoxazole (TMS), erythromycin, chloramphenicol, and rifampin was found in 194 isolates (41.5), 32 (6.8), 13 (2.8) and 3 (0.6), respectively. No isolates resistant to vancomycin were detected. The most commonly combined resistance patterns were: penicillin/TMS (35.6) and penicillin/TMS/cefotaxime (11.8). This study highlights the increased rate of drug resistant S. pneumoniae during the last years, and the importance of antimicrobial resistance surveillance of adequate empirical therapy involving local and regional susceptibility patterns.(AU)

Streptococcus pneumoniae: evolución de la resistencia a los antimicrobianos en un hospital de niños de Córdoba, Argentina / Streptococcus pneumoniae: evolution of antibiotic resistance in a pediatric hospital in Córdoba, Argentina

The wide variety of prevalence of antimicrobial resistant Streptococcus pneumoniae in different countries confirms the importance of determining local patterns of resistance. From 1992 to 2000, we studied the pattern of antimicrobial resistance in S. pneumoniae and its evolution along the years, using 468 strains isolated in the Hospital de Niños de Córdoba. A total of 177 isolates (37.8) were not susceptible to penicillin, with 19 intermediate and 18.8 resistant strains. High and intermediate resistance levels to cefotaxime were 4.9 and 10.9, respectively. Decreased susceptibility to trimethoprim/sulfamethoxazole (TMS), erythromycin, chloramphenicol, and rifampin was found in 194 isolates (41.5), 32 (6.8), 13 (2.8) and 3 (0.6), respectively. No isolates resistant to vancomycin were detected. The most commonly combined resistance patterns were: penicillin/TMS (35.6) and penicillin/TMS/cefotaxime (11.8). This study highlights the increased rate of drug resistant S. pneumoniae during the last years, and the importance of antimicrobial resistance surveillance of adequate empirical therapy involving local and regional susceptibility patterns.

Specific IgA detection can be used for perinatal diagnosis of HIV in children under protocol ACTG 076.

Detection of anti-HIV-1 IgA antibodies using a modified ELISA test for the early diagnosis of perinatally acquired HIV-1 infection in children treated with protocol ACTG 076 was evaluated. A total of 177 sera were obtained from 141 infants between 1 and 12 months of age (46 were treated and 95 were non-treated with protocol ACTG 076) and tested for HIV IgA antibodies by an ELISA test after removal of IgG with recombinant protein G. Infants were classified according to CDC's classification system after a follow-up until 20 months of age. Of the 46 treated children 22 turned out to be infected and in the group of 95 untreated children, 52 were infected. All 81 samples from uninfected children treated or untreated with protocol ACTG 076 were persistently IgA-negative. HIV IgA antibodies were detected in 14 of 25 plasma samples from infected children with treatment, and in 58 of 71 samples in infected children without treatment. Considering that the sensitivity of this test is lower in children younger than 6 months the population of children studied was divided into two groups; those under and those over 6 months of age. No significant differences were observed in the detection of IgA in treated or untreated children in both age groups. The overall specificity of the test was 100 per cent; sensitivity in children older than 6 months was 76.92 per cent in treated children and 93.10 per cent in untreated children. In spite of the small number of samples studied it could be demonstrated that treatment with zidovudine does not affect the detection of IgA antibodies. This is a simple and inexpensive method that could be used for diagnosis of treated and untreated children in developing countries.

Streptococcus pneumoniae: evolución de la resistencia a los antimicrobianos en un hospital de niños de Córdoba, Argentina. / [Streptococcus pneumoniae: evolution of antibiotic resistance in a pediatric hospital in Córdoba, Argentina]

The wide variety of prevalence of antimicrobial resistant Streptococcus pneumoniae in different countries confirms the importance of determining local patterns of resistance. From 1992 to 2000, we studied the pattern of antimicrobial resistance in S. pneumoniae and its evolution along the years, using 468 strains isolated in the Hospital de Niños de Córdoba. A total of 177 isolates (37.8

) were not susceptible to penicillin, with 19

intermediate and 18.8

resistant strains. High and intermediate resistance levels to cefotaxime were 4.9

and 10.9

, respectively. Decreased susceptibility to trimethoprim/sulfamethoxazole (TMS), erythromycin, chloramphenicol, and rifampin was found in 194 isolates (41.5

), 32 (6.8

), 13 (2.8

) and 3 (0.6

), respectively. No isolates resistant to vancomycin were detected. The most commonly combined resistance patterns were: penicillin/TMS (35.6

) and penicillin/TMS/cefotaxime (11.8

). This study highlights the increased rate of drug resistant S. pneumoniae during the last years, and the importance of antimicrobial resistance surveillance of adequate empirical therapy involving local and regional susceptibility patterns.

[Evaluation of indirect immunofluorescence as a supplementary test for the diagnosis of HIV-1 infection]. / Evaluación de la inmunofluorescencia indirecta como prueba suplementaria para el diagnóstico de la infección por HIV-1.

In order to be used as an alternative or complementary test to confirm HIV-1 infection, the efficiency of indirect immunofluorescence assay (IFA) was compared with Western blot (WB) in 362 samples from persons with high and low risk behaviour. A panel of sera with 220 WB positive, 122 WB negative and 20 WB indeterminate sera were tested by an "in house" IFA. The sensitivity of IFA was found to be 98.63% and the specificity 98.36%. Therefore, IFA appeared to be an efficient alternative method to WB, since the cost of testing by IFA is less than 10% of WB testing. We observed a direct relationship between WB protein reactivity and IFA results. In 15 samples with coincident indeterminate results for WB and IFA, antibody reactivity to p24 and gp160 presented the highest frequency. On the other hand, antibodies to viral glycoproteins were always present in IFA weak positive samples, showing their high predictive value.

Evaluación de la inmunofluorescencia indirecta como prueba suplementaria para el diagnóstico de la infección por HIV-1 / Evaluation of indirect inmunofluorescence as a confirmatory test for the diagnosis of HIV-1 infection

Se comparó la eficiencia diagnóstica de la inmunofluorescencia indirecta (IFI) como método confirmatorio de la infección por HIV-1 en muestras de suero de 362 personas con conductas de alto y bajo riesgo. El panel compuesto por 220 positivos, 122 negativos y 20 indeterminados por Western blot (WB) fue ensayado por una técnica de IFI desarrollada en nuestro laboratorio. La sensibilidad calculada fue 98,63 por ciento y la espicificidad 98,36 por ciento, indicando que la IFI es un método alternativo para la confirmación de la presencia de anticuerpos contra el HIV-1. Dado que su costo es menor que el 10 por ciento comparado con el del WB, se justifica su introducción en el algoritmo de diagnóstico serológico de HIV-1. Se observó también una relación directa entre la reactividad de las proteínas del WB y los resultados de IFI. En 15 muestras con resultado indeterminado por WB e inespecífico por IFI, las bandas más observadas fueron la p24 seguida de la gp160; por otro lado los anticuerpos contra las glicoproteínas virales son los que presentan mayor frecuencia en las muestras positivas débiles, demostrando su alto valor predictivo (AU)

Evaluación de la inmunofluorescencia indirecta como prueba suplementaria para el diagnóstico de la infección por HIV-1 / Evaluation of indirect inmunofluorescence as a confirmatory test for the diagnosis of HIV-1 infection

Se comparó la eficiencia diagnóstica de la inmunofluorescencia indirecta (IFI) como método confirmatorio de la infección por HIV-1 en muestras de suero de 362 personas con conductas de alto y bajo riesgo. El panel compuesto por 220 positivos, 122 negativos y 20 indeterminados por Western blot (WB) fue ensayado por una técnica de IFI desarrollada en nuestro laboratorio. La sensibilidad calculada fue 98,63 por ciento y la espicificidad 98,36 por ciento, indicando que la IFI es un método alternativo para la confirmación de la presencia de anticuerpos contra el HIV-1. Dado que su costo es menor que el 10 por ciento comparado con el del WB, se justifica su introducción en el algoritmo de diagnóstico serológico de HIV-1. Se observó también una relación directa entre la reactividad de las proteínas del WB y los resultados de IFI. En 15 muestras con resultado indeterminado por WB e inespecífico por IFI, las bandas más observadas fueron la p24 seguida de la gp160; por otro lado los anticuerpos contra las glicoproteínas virales son los que presentan mayor frecuencia en las muestras positivas débiles, demostrando su alto valor predictivo