Rosa García

Fractionation and Biological Characterization of Olea europea Pollen Extract

Preliminary fractionation of Olea european pollen extract has been performed. At least 10 antigenic fractions have been found by crossed electrophoresis. After Sephadex gel filtration, two fractions with a molecular weight of 160,000 and 65,000 have been obtained. The fractions were evaluated for allergenic activity by two in vitro techniques: polystyrene tube radioimmunoassay (PTRIA) and basophil degranulation test (BDT), and by skin tests. All tests indicated that the most reactive fractions were those in the 65,000 molecular weight peak. BDT has been shown to be a very reliable method as compared with histamine release and other parameters. Although common antigenic fractions have been found for Lollium perenne and O. europea, no cross-allergenicity has been shown by PTRIA inhibition.

Long-Term Follow-Up of Asymptomatic HIV-Infected Patients Who Discontinued Antiretroviral Therapy

BACKGROUND Whether asymptomatic human immunodeficiency virus (HIV)-infected patients can interrupt treatment remains unknown. METHODS We performed a prospective, observational study of 46 patients who started therapy with >300 CD4+ cells/mm3 and/or <70,0000 HIV-1 RNA copies/mL. Patients had been receiving highly active antiretroviral therapy (HAART) for at least 6 months. HAART was discontinued, and plasma HIV-1 RNA loads and CD4+ cell counts were determined at 4-month intervals. RESULTS At the time of HAART discontinuation, the median CD4+ cell count was 793 cells/mm3, and all patients had undetectable viral loads. A rapid decrease of 173 cells/mm3 in the median CD4+ cell count was observed during the first 4 months after HAART was stopped, followed by a slower decrease of 234 cells/mm3 between months 5 and 20. The decrease in the median CD4+ cell count early after HAART discontinuation was inversely correlated with the increase that occurred during receipt of therapy (r=-0.653) and with the count at the time of HAART discontinuation (r=-0.589). The decrease in the median CD4+ cell count after the fourth month without HAART was correlated with the nadir count before HAART initiation (r=-0.349) and the increase during treatment (r=-0.322). The median follow-up duration was 20 months. After 12, 24, and 36 months of observation, 33 patients (71.7%), 22 patients (47.8%), and 16 patients (34.7%), respectively, remained free of therapy. Adverse clinical events were not seen, and all patients who reinitiated HAART responded rapidly. CONCLUSION Selected asymptomatic HIV-infected patients can safely discontinue therapy for prolonged periods of time.

Long-Term Efficacy and Safety of Protease Inhibitor Switching to Nevirapine in HIV-Infected Patients with Undetectable Virus Load

BACKGROUND Simplified highly active antiretroviral therapy (HAART) regimens are becoming widely used, particularly as a result of the side effects of and difficult compliance with protease inhibitor (PI) therapy. However, the long-term efficacy of HAART has not been properly assessed. METHODS We performed a prospective study of 110 patients infected with human immunodeficiency virus type 1 (HIV-1) with undetectable virus load who discontinued PI therapy and initiated therapy with nevirapine without changing nucleoside analogues. Reasons for switching were treatment simplification (45%), lipodystrophy (24%), renal problems (23%), and dyslipidemia (8%). HIV-1 load, CD4 cell count, and fasting biochemistry profiles were performed at the time of switching (baseline) and every 3-4 months thereafter. The aim of the study was to evaluate the long-term efficacy and safety of this combination. RESULTS Sixty-eight patients (61.8%) had a duration of follow-up of 3 years. The mean increase in the CD4 cell count after 3 years was 90 cells/microL (13.8% from baseline). Virus loads remained undetectable in all patients but 9 (8.2%). Triglyceride levels dramatically improved at 12 months (a 75% decrease; P<.02) and remained statistically significant over time (P<.04). The same occurred with serum cholesterol levels: there was an initial reduction of 25% (P<.02) and at the end of the follow-up period (P<.015). However, at the long-term evaluation, complete normalization of mean serum cholesterol and triglyceride levels could not be achieved. Sixteen patients (14.5%) had to stop therapy as a result of nevirapine-associated side effects. CONCLUSIONS The switching of a PI to nevirapine is a safe and well-tolerated option for maintaining long-term virological suppression and immunological control. Three years after starting nevirapine therapy, rates of hypercholesterolemia and hypertriglyceridemia improved, although normal cholesterol and triglyceride values were not achieved.

Quantification of the Major Allergen from Cypress (Cupressus arizonica) Pollen, Cup a 1, by Monoclonal Antibody-Based ELISA

Background: Cypress pollen allergy is an important cause of rhinoconjunctivitis and asthma in Mediterranean countries. Cypress allergenic extracts are difficult to produce since they have low protein and high carbohydrate content, thus accurate standardization of them is essential to guarantee their quality. The aim of this study is to develop a sandwich ELISA for the quantification of Cup a 1, the major allergen of cypress (Cupressus arizonica) pollen extract. Methods: Monoclonal antibodies directed to purified Cup a 1 were produced. Two of them (9C7 as capture antibody and 3D2 as the tracer) were selected to develop a quantitative sandwich ELISA. This ELISA was subsequently evaluated and compared with other techniques. Results: The described ELISA is very sensitive with a detection limit of 8.7 ng/ml and a practical working range of 62.5–1,000 ng/ml. The assay is also highly reproducible with intra-assay and interassay coefficients of variation of less than 10%. The purified Cup a 1, used as standard, presents pectate lyase enzymatic activity. The assay also detected Cup a 1-like proteins in pollen from other Cupressaceae. A good correlation was obtained between Cup a 1 content of 12 C. arizonica pollen extracts and their IgE-binding activity. Conclusions: The described Cup a 1 ELISA is sensitive, specific and reproducible and can be used for the quantification of Cup a 1 in C. arizonica and other related pollen extracts. It also provides a reliable indication of the allergenic activity of the whole cypress pollen extract.

Clinical Characteristics and Outcome of Infective Endocarditis in Individuals of the General Population Managed at a Teaching Hospital Without Cardiac Surgery Facilities. Study of 120 Cases

INTRODUCTION AND OBJECTIVES To assess the clinical characteristics and management of infective endocarditis at a teaching hospital without cardiac surgery facilities. METHODS Descriptive case-control study looking at trends. Risk factors, the occurrence of complications, the rate of referral for cardiac surgery, and the mortality rate were assessed. RESULTS The study included 120 patients referred between 1990 and 2004, with a mean age of 50.8 (17.8) years (67.6% men). Disease incidence did not change throughout the study. Some 55% of infective endocarditis cases were from the ordinary general population, 25% were intravenous drug users, and 20% were of nosocomial origin. The number in the last category had increased over time. The most commonly isolated microorganism was Staphylococcus aureus. Around 83% of patients presented with a severe complication, with cardiac failure and septic metastasis being the most common. The in-hospital mortality rate was 19.2%. Acute renal failure (odds ratio 6.7, 95% confidence interval, 1.9-24) and perivalvular abscess (odds ratio 9.2, 95% confidence interval, 1.6-54) were independent predictors of death. The introduction in 2002 of a multidisciplinary infective endocarditis team, which included a consultant cardiac surgeon, was associated with a significant increase in referrals for surgery, from 14.5% to 34.5% (P=.03), though in-hospital mortality was not significantly altered, decreasing from 20.9% to 13.8% (P=.4). CONCLUSIONS The occurrence of acute renal failure and perivalvular abscess worsen the prognosis of infective endocarditis. The introduction of a multidisciplinary infective endocarditis team altered management of the disease and increased referrals for cardiac surgery.

Conocimientos sobre los procedimientos correctos de medición de la presión arterial entre estudiantes universitarios de ciencias de la salud

Este trabajo describe los conocimientos sobre los procedimientos correctos de medicion de la presion arterial (PA) entre 175 estudiantes de tercer curso y 176 de sexto curso de una facultad de medicina y 58 de tercer curso de una escuela de enfermeria. Solo el 51,8% de los estudiantes consideraban que sabian medir correctamente la PA (el 28,6% en tercero de medicina, el 61,9% en sexto y el 91,4% en enfermeria). En sexto de medicina, el 12,5% conocia el tamano apropiado del manguito; el 35%, que el diafragma del estetoscopio no debe introducirse debajo del manguito, y el 43%, que la velocidad de desinflado del manguito influye en la PA. En enfermeria, el 33% conocia el tamano adecuado del manguito; el 22%, que la PA ambulatoria debe medirse mas de una vez en cada visita, y el 55%, que el diagnostico de hipertension requiere tener la PA elevada en mas de una visita.

Knowledge of correct blood pressure measurement procedures among medical and nursing students.

This study reports on the level of knowledge about correct blood pressure (BP) measurement procedures among 175 third-year and 176 sixth-year students at a medical faculty and 58 third-year students at a nursing college. Only 51.8% of all students thought that they knew how to measure BP correctly (28.6% in the third year and 61.9% in the sixth year at the medical faculty and 91.4% at the nursing college). Among sixth-year medical students, 12.5% knew the appropriate cuff size, 35% that the stethoscope diaphragm should not be placed under the cuff, and 43% that the cuff deflation rate affects BP measurement. Among nursing students, 33% knew the appropriate cuff size, 22% that an outpatients BP should be measured more than once during each visit, and 55% that a diagnosis of hypertension can only be made if the BP is elevated on more than one visit.

Immune complexes from HIV-1+ patients contain infectious virus able to infect normal lymphocytes

It has been suggested that circulating immune complexes containing HIV-1 could be involved in enhancement of the infection through Fc receptors. To test this hypothesis, immune complexes precipitated from the sera of 25 HIV-1-seropositive individuals, at different stages of the disease, were assayed for the presence of infectious virus. When added to phytohemagglutinin-activated peripheral blood lymphocytes, seven of 25 complexes were able to sustain a productive infection. This was demonstrated by release of HIV-p24 antigens in the supernatant of activated but not resting peripheral blood lymphocytes after several days of culture. Moreover, peripheral blood lymphocytes cultured with complexes from HIV-1+ patients but not from control subject, contained integrated HIV-1 provirus in their DNA. These results demonstrated the presence of infectious virus in immune complexes from HIV-1+ patients.

Occupational allergic multiorgan disease induced by wheat flour

Bakers are repeatedly exposed to wheat flour (WF) and may develop sensitization and occupational rhinoconjunctivitis and/or asthma to WF allergens.1 Several wheat proteins have been identified as causative allergens of occupational respiratory allergy in bakery workers.1 Testing of IgE reactivity in patients with different clinical profiles of wheat allergy (food allergy, wheat-dependent exercise-induced anaphylaxis, and bakers asthma) to salt-soluble and salt-insoluble protein fractions from WF revealed a high degree of heterogeneity in the recognized allergens. However, mainly salt-soluble proteins (albumins, globulins) seem to be associated with bakers asthma, and prolamins (gliadins, glutenins) with wheat-dependent exercise-induced anaphylaxis, whereas both protein fractions reacted to IgE from food-allergic patients.1 Notwithstanding, gliadins have also been incriminated as causative allergens in bakers asthma.2 We report on a 31-year-old woman who had been exposed to WF practically since birth because her family owned a bakery housed in the same home where they lived. She moved from this house when she was 25 years, but she continued working every day in the family bakery. In the last 8 years she had suffered from work-related nasal and ocular symptoms such as itching, watery eyes, sneezing, nasal stuffiness, and rhinorrhea. These symptoms markedly improved when away from work and worsened at work. In the last 5 years, she had also experienced dysphagia with frequent choking, especially when ingesting meats or cephalopods, which had partially improved with omeprazole therapy. Two years before referral to our clinic, she began to have dry cough and breathlessness, which she also attributed to her work environment. Upper and lower respiratory tract symptoms increased when sifting the WF and making the dough. The patient did not experience gastrointestinal symptoms with ingestion of cereal products. Skin prick test results were positive to grass (mean wheal, 6 mm), cypress (5 mm) and Russian thistle pollen (4 mm), WF (4 mm), and peach lipid transfer protein (6 mm) and were negative to rice flour, corn flour, profilin, mites, molds, and animal dander. Skin prick test with a homemade WF extract (10% wt/vol) was strongly positive (15 mm). Serologic tests yielded the following results: eosinophil cationic protein, 47 ?g/L; total serum IgE, 74 kU/L; specific IgE (ImmunoCAP; ThermoFisher, Uppsala, Sweden) to WF, 7.4 kU/L; barley flour, 1.24 kU/L; and corn, gluten, alpha-amylase, peach, and apple, less than 0.35 kU/L. Specific IgE binding to microarrayed purified WF allergens (WDAI-0.19, WDAI-0.53, WTAI-CM1, WTAI-CM2, WTAI-CM3, WTAI-CM16, WTAI-CM17, Tri a 14, profilin, ?-5-gliadin, Tri a Bd 36 and Tri a TLP, and gliadin and glutamine fractions) was assessed as described elsewhere.3 The patients serum specifically recognized ?-5-gliadin and the gliadin fraction, and no IgE reactivity was observed to other wheat allergens. Spirometry revealed a forced vital capacity of 3.88 L (88%), an FEV1 of 3.04 L (87%), and FEV1/forced vital capacity of 83%. A methacholine inhalation test was performed following an abbreviated protocol,4 and the results were expressed as PD20 in cumulative dose (mg) of methacholine. Methacholine inhalation challenge test result was positive (0.24 mg cumulative dose) when she was working, and after a 3-month period away from work and with no visits to the bakery house, it gave a negative result. A chest x-ray was normal. Specific inhalation challenge test was carried out in the hospital laboratory by tipping WF from one tray to another for 15 minutes. Spirometry was performed at baseline and at 2, 5, 10, 15, 20, 30, 45, and 60 minutes after the challenge with WF. Peak expiratory flow was measured at baseline and then hourly over 24 hours (respecting sleeping time). A 12% fall in FEV1 was observed at 20 minutes and a 26% drop in peak expiratory flow at 9 hours after exposure to WF,

Anti-CD4 Activity in Circulating Immune Complexes in HIV-lnfected Patients

Levels of circulating immune complexes (CIC) measured by precipitation with 1.04 M ammonium sulfate ranged from 22 to 2,040 micrograms/ml in a group of 141 HIV-infected patients. CIC were elevated (> 200 micrograms/ml) in 72.2% of infected individuals. When analyzed for their HIV antigen composition, those CIC containing HIV antigens were found more frequently in patients clinically affected (68.6%) than in asymptomatic individuals (31.4%; p < 0.001). Anti-CD4 activity of 89 isolated CIC was detected in 43.8% of these patients, but only in 7.6% of the cases these CIC could bind to native CD4+ molecules. CIC with anti-CD4 activity could inhibit PHA stimulation of normal peripheral blood lymphocytes. Anti-CD4 activity in CIC was independent of the clinical and immunological status of HIV-infected patients.