IFN-γ expression in placenta is associated to resistance to Chlamydia abortus after intragastric infection.

Intragastric infection mimics the natural route of infection of Chlamydia abortus (etiological agent of ovine enzootic abortion). In the mouse model, intragastric experimental infection induces very mild signs of infection followed by late term abortions, as it is shown by the natural ovine host. In order to evaluate the immune mechanisms associated to the dissemination of the pathogen from the gastrointestinal tract, we have administered an intragastric dose of C. abortus to pregnant mice. Systemic and local expression of cytokines, tissue colonization and excretion of bacteria after parturition were monitored during pregnancy. Susceptible CBA/J mice showed a higher bacterial colonization of the placenta and excretion of live bacteria after parturition that were related to a higher local IL-10 expression. By contrast, resistant C57BL/6 mouse strain had higher local IFN-γ mRNA expression in the placenta just before parturition and a transient bacterial colonization of the reproductive tract, with no excretion of C. abortus after parturition. In summary, intragastric infection not only mimics the natural route of infection of C. abortus, but can also be useful in order to understand the immunopathogenesis of chlamydial abortion in the mouse.

[Respiratory infections research: a perspective from the tuberculosis and respiratory infections area (TIR)]. / Investigación en infecciones respiratorias: una perspectiva desde el Area Tuberculosis e Infecciones Respiratorias (TIR).

The scientific production of the TIR Area of SEPAR during 2008 is reviewed. In pneumonias, studies on C-reactive protein, procalcitonin and the cytokines as predictive markers of treatment failure are noteworthy, as well as research into the genetic predisposition of the host (polymorphisms of mannose binding lectin) in the prognosis. Among the different activities on tuberculosis in the SEPAR <> year, was the publication of the new SEPAR guidelines for the <>. The studies into tuberculosis have been on, the tuberculosis infection, the new in vitro techniques for detecting interferon gamma, new non-bacillary tuberculosis diagnostic committees, and treatment schemes without rifampicin and isoniazid. In COPD,we have highlighted new aspects in the indications for antibiotic treatment in the Consensus Document for the antibiotic treatment of acute exacerbations of COPD, and in the SEPAR-ALAT Clinical Guidelines. In the field of cystic fibrosis (CF), we highlight 3 studies: a) association between colonising- Pseudomonas aeruginosa induced chronic infection and bronchial hyperreactivity; b) serum immunoglobulins response to Aspergillus fumigatus and Candida albicans in the colonising of the lower respiratory tract and its clinical significance; and c) prevalence of environmental mycobacteria in these patients. In the chapter on bronchiectasis, a study on the relationship between systemic inflammation and severity parameters is highlighted, and finally, the main contributions of the new SEPAR guidelines on the diagnosis and treatment of bronchiectasis.

Direct activation of telomerase by GH via phosphatidylinositol 3'-kinase.

Telomerase is a ribonucleoprotein DNA polymerase that has been associated with cell proliferation, cell survival and apoptosis inhibition. Telomerase is regulated by specific growth factors, cytokines and hormones. The present study examines the effect of GH on telomerase activity and identifies the signal transduction pathway involved in this process in Chinese hamster ovary (CHO)4 cells, which express rat GH receptor cDNA. Telomeric repeat amplification protocol assays demonstrated that treating CHO4 cells with increasingly high doses of GH up-regulated telomerase activity with the maximum activation at 24 h. Similarly, GH activated telomerase in another cell system, primary cultures of rat hepatocytes. The telomerase activation in CHO4 cells was produced with an increase in hamster telomerase catalytic subunit (hamTERT) mRNA expression. The telomerase activity induced by GH was specifically blocked by the phosphatidylinositol 3'-kinase (PI3-K) inhibitor, LY294002, but not by the MAP kinase kinase inhibitor, PD98059. These findings suggest that GH could activate telomerase through the direct activation of TERT transcription, as well as through the PI3-K signalling pathway.

Immunogenicity of a yeast-derived hepatitis B vaccine in hemodialysis patients.

In a multicenter study of hemodialysis patients in Spain, the immunogenicity of a yeast-derived recombinant deoxyribonucleic acid hepatitis B vaccine was evaluated. Two different vaccination schedules were examined: zero, one, two, six months and zero, one, two, 12 months. Two different dose levels (20 micrograms and 40 micrograms) were also compared. No serious adverse effects were reported by any of the vaccinees; the most frequently reported reaction was soreness at the injection site. This study also indicated that higher concentrations of antibodies are attained when more frequent doses of vaccine are administered. The yeast-derived vaccine produced an immune response similar to that of the plasma-derived vaccines.

Enriched protein diet-modified ghrelin expression and secretion in rats.

Gastrointestinal (GI) integrity and function are regulated by nutrition and growth factors. The discovery of ghrelin, a natural growth hormone (GH) secretagogue produced by the gastrointestinal (GI) tract, is a potential link between diet and growth signals. The aim of this study was to evaluate macronutrient effect on ghrelin expression and secretion in addition to some possible function in intestinal trophic status. Wistar rats were fed a high-carbohydrate, high-protein (HP), high-fat or standard (St) diet. Animals received the same daily food volume and caloric intake. After 7 days, animals were fasted for 24 h and blood and tissue samples were obtained just before feeding or at 2 or 6 h after feeding. Fasting high-protein-fed rats had higher ghrelin plasma levels than with rats fed the high-carbohydrate, high-fat or standard diets. Two-hours after refeeding, ghrelin plasma levels had decreased in all groups with a slight recovery at 6 h after refeeding, except in the high-protein group. Ghrelin plasma levels in rats fed with the high-protein diet correlated negatively with their GH and insulin-like growth factor 1 (IGF-1) plasma concentrations which were also the lowest among the study groups. In conclusion, ghrelin secretion was nutritionally manipulated because a protein-enriched diet increased its levels.

Diagnostic value of cytokeratin fragment 19 (CYFRA 21-1) in bronchoalveolar lavage fluid in lung cancer.

The aim of this study was to evaluate the diagnostic value of a new tumour marker, cytokeratin fragment 19 (CYFRA 21-1), in bronchoalveolar lavage fluid (BALF) for the diagnosis of lung cancer. The cross-sectional study included 36 patients with lung cancer, 19 with benign lung diseases and 13 control subjects. In the group with cancer, BAL was performed in the cancer-involved lung and in the opposite lung. Results in BALF were expressed both as absolute concentrations (ng ml-1) and referred to total protein (TP) (ng mg-1 TP), and results in plasma were expressed in ng ml-1. In BALF, there was no significant different between cancer and control groups. Using the 95th percentile of levels obtained in benign lung disease in BALF (specificity 95%) as the cut-off point, the sensitivity of CYFRA 21-1 was 13%. Positive and negative predictive values (PPV and NPV) at different pretest probabilities, and positive and negative gains were obtained applying a Bayesian analysis. Results showed low positive gains for PPV (maximal increase of 22%) and almost none for NPV (negative gains < 5%). In plasma, CYFRA 21-1 provided a sensitivity of 65%. The combination of BALF and plasma tumour marker levels showed a sensitivity of 69%. Therefore, measurement of CYFRA 21-1 in BALF has poor diagnostic value in lung cancer.

[Gain, loss and agreement between respiratory specialists and generalists in the diagnosis of asthma]. / Ganancia, pérdida y concordancia en el diagnóstico de asma entre neumólogos y no neumólogos.

OBJECTIVE: To determine and analyze the degree of agreement and disagreement in the diagnosis of bronchial asthma (BA) by respiratory disease specialists and generalists in regional hospital and primary care settings. MATERIAL AND METHODS: Ninety-six outpatients (16 to 70 years of age) were studied; all had been assigned a diagnosis of BA by the referring physician or by the respiratory disease specialist. We recorded 1) clinical symptoms, determining the initial probability of a diagnosis (IPD)of BA to be high, medium or low; 2) results of spirometry and bronchodilator testing (BDT), peak flow variability and methacholine challenge testing; 3) prick test results, eosinophil levels and total serum IgE levels. Three diagnoses were recorded: the initial diagnosis (ID) by the referring physician to whom follow-up data were unavailable; diagnosis by the respiratory disease specialist based only on clinical symptoms (RSS); and the final diagnosis(FD). To arrive at a FD of BA, it was necessary to have a high or medium IPD and a positive BDT. A Kappa test was used to analyze the degree of agreement among the three diagnoses. Group features associated with greater or lesser agreement were analyzed by chi-square tests and analysis of variance. RESULTS: Agreement was acceptable between RSS and FD (K = 0.63) but very low between ID and RSS and between ID and FD. In the latter two cases, agreement was greatest for patients diagnosed in hospital and for those with high IgE levels (p < 0.05), with high IPD, longer course of disease and a history of asthma (p < 0.01) (odds ratio =59.8). Diagnostic disagreement occurred mainly for patients for whom a BA diagnosis was gained later, the of under-diagnosis being 39%. The patients involved visited the physician only because they had observed an isolated symptom related to asthma (odds ratio = 119) and to arrive at a diagnosis bronchomotor tests other than BDT were required (p < 0.01). CONCLUSIONS: a) The degree of agreement for a diagnosis of BA is low. b)The functional profile of patients for whom diagnostic agreement exists differs from that of patients for whom diagnosis is gained through testing. c) In the context of this study, a high rate of under-diagnosis is evident.

[Profile of a pneumology regional health service]. / Perfil de una consulta de neumología de ámbito comarcal.

This study analyzes patient demand in a regional public health pulmonology practice. The following data were recorded for all first-visit patients for a period of two years: age, sex, referral source, initial diagnosis by the referring physician, final diagnosis by the pulmonologist, and destination. The service studied 1,486 patients (men/women: 1.5). Most (71%) were between 40 and 80 years old. Referrals were from the family doctor (60%), health center (9%), emergency service (10%), hospital (12%), other specialists (6%), and others (3%). The most frequent reasons for remission were upper airway disease (UAD) (36%), specifically chronic obstructive pulmonary disease (COPD) and asthma, and the presentation of symptoms (28%) such as dyspnea, cough, hemoptysis and chest pain. Analysis of the final diagnoses for the patients presenting with symptoms showed that no disease could be detected in one third of those with dyspnea and hemoptysis or in half of those who complained of chest pain; acute respiratory infection was diagnosed in 45% of those complaining of persistent cough. UAD was the most frequent cause of symptoms. The index of doctor's visit/inhabitant was 0.97% for patients referred by family doctors and 0.38% for those from health centers, but the initial and final diagnosis profiles of these patients were not statistically different. Patients referred by emergency services had significantly more (p < 0.001) in number of radiological findings and hemoptysis. Those sent from hospitals more often suffered pneumonia. In conclusion, this profile of a regional public health pulmonology practice shows that: 1) UAD and clinical symptoms are the most frequent reasons for patient remission; 2) family doctors generate three times mor demand for services than do health centers, and 3) 14% of patients can be considered normal.

[Descriptive study of patients diagnosed with asthma in a regional respiratory medicine practice]. / Estudio descriptivo de los pacientes diagnosticados de asma en una consulta neumológica de ámbito comarcal.

OBJECTIVE: To describe the characteristics of patients diagnosed of bronchial asthma (BA) in a regional respiratory medicine practice. METHODS: Over a period of two years, 88 adult patients followed a prospective-diagnostic protocol for BA that included taking of patient history to determine the probability of initial diagnosis (PID) of asthma, assessment of atopy and a lung function test that included spirometry with a bronchodilator test, recording of forced expiratory volume, and a methacholine challenge test. BA was diagnosed when symptoms denoting high or moderate PID were present and there was a positive reversibility and/or bronchial hyperreactivity test. The severity of disease was evaluated using the consensus criteria of the Global Initiative for Asthma (GINA). A student-t test and a chi 2 test were performed to compare data. RESULTS: BA was diagnosed in 24 men (30%) and 56 women (70%) with a mean age of 43.0 +/- 17.6 years (range 16 to 68). Significant differences between men and women were detected only for function parameters and cigarette consumption (p < 0.05). Fifty-three patients (66%) were referred by general practitioners, 22 (27%) were referred by hospital, and 5 came from other sources. The first group had a shorter history of disease course (p = 0.05) upon first evaluation. Sixty-one patients (76%) had mild asthma; these patients were younger and had had symptoms for a shorter period of time than those with more serious degrees of disease (16 with moderate asthma and 3 with severe asthma) (p < 0.05). Symptoms indicated a high PID in 57 cases (71%), and this figure increased significantly to 89.5% for those with more severe asthma (p < 0.05). Asthma was intrinsic for 39 patients and extrinsic for 41, with significant differences in age and total IgE between the two groups (p < 0.01). CONCLUSIONS: 1. BA is more than twice as common among women as among men. 2. Two thirds of patients are referred by general practitioners. 3. Mild asthma clearly predominates. 4. Clinical symptoms point to asthma, the PID in most cases being high.

[Recombinant hepatitis B vaccine in health personnel. Immunogenicity of a rapid vaccination schedule]. / Vacuna recombinante de la hepatitis B en personal sanitario. Inmunogenicidad de una pauta rápida de vacunación.

The immunogenic effect of a recombinant hepatitis B vaccine, administered in a dosage of 20 micrograms intramuscularly in the deltoid muscle on the months 0, 1 and 2, was evaluated in 185 employers of a general hospital. The influence of sex, age, overweight and smoking habit on the antibody response induced by the vaccine was also assessed. The seroconversion rate 40-60 days after the third dose in the 160 health professionals who completed the vaccination schedule was 88% (83% in males and 90% in females). It was 97% in individuals less than 30 years of age, 89% in those between 30 and 50 years, and 69% in those over 50 years. It was 97% in non obese and 66% in obese individuals (p less than 0.0001). No differences in seroconversion rate were found between nonsmokers, moderate smokers and heavy smokers. Untoward reactions were minimal. The rate of vaccinated individuals developing anti-HBs titers higher than 10 mIU/ml was 68%. This rate was lower than that found in most studies after vaccination in the months 0, 1 and 6, the difference being probably due to the use of a rapid schedule with a short interval between the second and third doses. Nevertheless, this schedule facilitates the vaccination programs in health staff and reduces the failure rate due to poor compliance.

[Epidemic Kaposi's sarcoma. A study of 5 cases]. / Sarcoma de Kaposi epidémico. Estudio de cinco casos.

5 cases of epidemic Kaposi's sarcoma in 5 homosexual males with HIV infection are presented. All of them died 3, 8, 9, 11, 24 months, respectively, after the initial diagnosis. We review the actual therapeutic possibilities of this neoplasia.

Effects of intravenous administration of allogenic bone marrow- and adipose tissue-derived mesenchymal stem cells on functional recovery and brain repair markers in experimental ischemic stroke.

INTRODUCTION: Stem cell therapy can promote good recovery from stroke. Several studies have demonstrated that mesenchymal stem cells (MSC) are safe and effective. However, more information regarding appropriate cell type is needed from animal model. This study was targeted at analyzing the effects in ischemic stroke of acute intravenous (i.v.) administration of allogenic bone marrow- (BM-MSC) and adipose-derived-stem cells (AD-MSC) on functional evaluation results and brain repair markers. METHODS: Allogenic MSC (2 × 106 cells) were administered intravenously 30 minutes after permanent middle cerebral artery occlusion (pMCAO) to rats. Infarct volume and cell migration and implantation were analyzed by magnetic resonance imaging (MRI) and immunohistochemistry. Function was evaluated by the Rogers and rotarod tests, and cell proliferation and cell-death were also determined. Brain repair markers were analyzed by confocal microscopy and confirmed by western blot. RESULTS: Compared to infarct group, function had significantly improved at 24 h and continued at 14 d after i.v. administration of either BM-MSC or AD-MSC. No reduction in infarct volume or any migration/implantation of cells into the damaged brain were observed. Nevertheless, cell death was reduced and cellular proliferation significantly increased in both treatment groups with respect to the infarct group. At 14 d after MSC administration vascular endothelial growth factor (VEGF), synaptophysin (SYP), oligodendrocyte (Olig-2) and neurofilament (NF) levels were significantly increased while those of glial fiibrillary acid protein (GFAP) were decreased. CONCLUSIONS: i.v. administration of allogenic MSC - whether BM-MSC or AD-MSC, in pMCAO infarct was associated with good functional recovery, and reductions in cell death as well as increases in cellular proliferation, neurogenesis, oligodendrogenesis, synaptogenesis and angiogenesis markers at 14 days post-infarct.

Functional recovery after hematic administration of allogenic mesenchymal stem cells in acute ischemic stroke in rats.

Hematic administration of bone marrow-derived mesenchymal stem cells (MSCs) in acute ischemic stroke may not only be an effective reparative treatment but also a brain protective therapy that improves neurological recovery. Our purpose was to study whether either i.v. or intracarotid (i.c.) administration of allogenic MSCs during the acute phase were effective in improving neurological recovery and decreasing brain damage in an experimental rat model. In a model of permanent middle cerebral artery occlusion (pMCAO), we analyzed: neurological evaluation; MSCs migration and implantation; interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) levels; lesion volume; cell death; cellular proliferation; vascular endothelial growth factor (VEGF) expression and blood vessel number. Regardless of the administration route, treated groups showed better neurological recovery, without significant differences between the two groups. Migration and implantation of MSCs in the lesion area was observed in animals receiving i.c. but not i.v. treatment. The highest cytokine values were observed in the i.v. MSCs and i.c. control groups, and these levels were significantly different from the corresponding i.v. control and i.c. MSCs groups, respectively. In addition, there were significant differences between the i.v. MSCs and i.c. MSCs groups in IL-6 levels. Neither treatment reduced infarction volume. However, cell death, measured as TUNEL+ cells was decreased with significant differences between control groups. BrdU+ cells were also significantly increased in the peri-infarct zone at 14 days. VEGF expression was significantly higher in the i.c. MSCs group than in the i.c. control group and blood vessel number was significantly higher in treated groups than control groups with significant differences in the peri-infarct zone at 14 days. We conclude that allogenic MSCs administration shows therapeutic efficacy in our acute ischemic stroke model. Both routes demonstrably improved neurological recovery and provided brain protection.

Fibronectin in bronchoalveolar lavage fluid in lung cancer: tumor or inflammatory marker?

OBJECTIVE: To evaluate the potential value of the level of fibronectin (FN) in the bronchoalveolar lavage fluid (BALF) as a lung tumor marker. METHOD: We compare the results of determinations in the tumor-bearing and tumor-free lungs of 38 patients with lung cancer, in 19 patients with benign lung diseases and in 13 healthy control subjects. FN was determined in BALF by a competitive ELISA and was also measured in plasma with a nephelometric assay. FN levels in BALF are expressed also referred to total protein content (ng FN/mg TP). RESULTS: The distribution of FN levels in BALF was not gaussian, and the levels in cancer patients were significantly greater than in healthy controls or in the contralateral lung. There was no significative difference in the levels in cancer compared with several benign conditions. In this study we observed a positive correlation between FN levels and total protein and polymorphonuclear leukocytes in BALF in the lung cancer and benign diseases group. CONCLUSION: These results do not support the possibility that FN levels in BALF were a general marker of lung cancer, and suggest that FN may mark the existence of pathologic lung processes associated with inflammation and, perhaps tissue repair after bronchopulmonary injury.

Repeated pulmonary infection by Nocardia asteroides complex in a patient with bronchiectasis.

A rare case of pulmonary nocardiosis was presented in a nonimmunocompromised patient who had chronic airway obstruction and bronchiectasis without corticoid treatment. The microbial diagnosis was established after isolating Nocardia in bronchial aspirate and sputum samples. An in vitro study showed sensitivity only to imipenem, netilmicine, amikacin and ofloxacin. The evolution was chronic, with multiple clinical recurrences in spite of prolonged antibiotic treatment. Finally, the eradication of Nocardia was achieved with the combination of imipenem and amikacin.

Prognostic factors in restoration of pulmonary flow after submassive pulmonary embolism: a multiple regression analysis.

Defects as evaluated by lung perfusion scans may persist even 6 months after pulmonary embolism (PE), when treatment is withdrawn. The aim of this study was to evaluate the effect of several potential factors on the resolution of lung perfusion defects, both during the first days and at 6 months, when patients were discharged. In a retrospective follow-up cohort study we included 102 patients with PE, diagnosed lung from a ventilation/perfusion (V'/Q') scan, following Prospective Investigation of Pulmonary Embolism Diagnosis (PIOPED) criteria, together with a phlebographic study of lower extremities or angiography. Lung perfusion scan was performed at diagnosis, and in two follow-up evaluations, at 7-10 days and at 6 months. Potential factors studied were: age; sex; presence of underlying cardiac or pulmonary disease; venous insufficiency; alveolar-arterial pressure difference for oxygen; delay in diagnosis; abnormalities in electrocardiogram or chest radiograph; and the size of defects as shown in lung perfusion scans. All factors were studied with regards to the size of the defects at the two follow-up evaluations, through a univariate statistical analysis and two multiple stepwise regression analysis. Multivariate statistical analysis selected four factors: size of defects at diagnosis; prior cardiopulmonary disease; delay in diagnosis; and sex, as synergistic variables to predict defect size at 7-10 days. On the other hand, the defect size at 7-10 days was the only variable selected as a predictor of the size of defects at 6 months. Resolution of pulmonary defects during the first days after diagnosis of pulmonary embolism is influenced by the initial defect size, prior cardiopulmonary diseases and sex. The size of residual defects at 6 months depends mainly on the size of defects at 7-10 days.

Impaired response to recombinant hepatitis B vaccine in HIV-infected persons.

We studied the immunogenicity of the standard schedule of recombinant hepatitis B vaccine (20 micrograms per dose at months 0, 1, and 6) in 21 anti-human immunodeficiency virus (HIV)-positive persons. Relatively low titers of anti-HBs developed in only five subjects (23.8%) 1 month after the third dose; all five had T4 cell counts greater than 700 cells/mm and none of the 11 subjects with a T4 cell count below this value responded. Five of the 16 nonresponders to the vaccine later had acquired immunodeficiency syndrome (AIDS)-related complex (two) and AIDS (three), while none of the responders did. Our results show that anti-HIV-positive persons are poor responders to the recombinant hepatitis B vaccine, and that the absence of a response is an indicator of a more severe immune deficiency and of a poor prognosis. An optimal regimen of hepatitis B vaccination in HIV-infected persons is still to be established.