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Saccharomyces cerevisiae is an important unicellular yeast species within the biotechnological and the food and beverage industries. A significant application of this species is the production of ethanol, where concentrations are limited by cellular toxicity, often at the level of the cell membrane. Here, we characterize 61 S. cerevisiae strains for ethanol tolerance and further analyze five representatives with various ethanol tolerances. The most tolerant strain, AJ4, was dominant in coculture at 0 and 10% ethanol. Unexpectedly, although it does not have the highest noninhibitory concentration or MIC, MY29 was the dominant strain in coculture at 6% ethanol, which may be linked to differences in its basal lipidome. Although relatively few lipidomic differences were observed between strains, a significantly higher phosphatidylethanolamine concentration was observed in the least tolerant strain, MY26, at 0 and 6% ethanol compared to the other strains that became more similar at 10%, indicating potential involvement of this lipid with ethanol sensitivity. Our findings reveal that AJ4 is best able to adapt its membrane to become more fluid in the presence of ethanol and that lipid extracts from AJ4 also form the most permeable membranes. Furthermore, MY26 is least able to modulate fluidity in response to ethanol, and membranes formed from extracted lipids are least leaky at physiological ethanol concentrations. Overall, these results reveal a potential mechanism of ethanol tolerance and suggest a limited set of membrane compositions that diverse yeast species use to achieve this. IMPORTANCE Many microbial processes are not implemented at the industrial level because the product yield is poorer and more expensive than can be achieved by chemical synthesis. It is well established that microbes show stress responses during bioprocessing, and one reason for poor product output from cell factories is production conditions that are ultimately toxic to the cells. During fermentative processes, yeast cells encounter culture media with a high sugar content, which is later transformed into high ethanol concentrations. Thus, ethanol toxicity is one of the major stresses in traditional and more recent biotechnological processes. We have performed a multilayer phenotypic and lipidomic characterization of a large number of industrial and environmental strains of Saccharomyces to identify key resistant and nonresistant isolates for future applications.

Resumen La miocardiopatia hipertrofica es la cardiopatia monogenica mas frecuente. Su expresion fenotipica es bastante variable. Hasta en un 60% de los casos se describen mutaciones en los genes que codifican las proteinas del sarcomero cardiaco. La secuenciacion masiva del acido desoxirribonucleico posibilita descubrir nuevos genes responsables de la enfermedad, pero tiene el inconveniente de descubrir numerosas variantes de significado incierto en estos pacientes. La estrategia ante las mismas, sobre todo cuando no se segregan con la enfermedad, es uno de los retos de la genetica. Los criterios de patogenicidad pueden ayudar a catalogar esa variante. Las pruebas geneticas al caso indice permiten realizar un diagnostico y la posibilidad de efectuarlo en cascada a los familiares de primer grado. La presencia, o no, de un genotipo positivo en los familiares determinara las pautas de seguimiento posteriores. La aparicion de un genotipo positivo empeora el pronostico, independientemente del tipo de mutacion.

espanolIntroduccion. Con la llegada a la universidad de una nueva generacion de alumnos, la ‘generacion Z’, considerada como la primera generacion nativa digital, se hace necesario implantar nuevas metodologias docentes en el ambito universita-rio para mejorar el proceso de ensenanza-aprendizaje y conseguir una mayor motivacion en el alumno. Para ello, el uso de nuevas tecnologias de la informacion y de la comunicacion ha permitido dinamizar estos procesos y aumentar la moti-vacion del alumnado en todos los niveles educativos, incluyendo la docencia universitaria. Sujetos y metodos. Utilizacion de la aplicacion educativa Kahoot a traves de dispositivos moviles, con los alumnos matri-culados de la asignatura de ‘Anatomia Humana II (Esplacnologia)’, de segundo curso del Grado de Medicina en la Univer-sidad de Zaragoza. Resultados. El impacto que la experiencia ha causado en los estudiantes se ha medido a traves de encuestas de valora-cion cualitativas, analizando sus resultados de aprendizaje. Estos resultados han sido valorados de manera muy positiva por el alumnado, tanto en lo referente a utilidad en la ensenanza-aprendizaje como a motivacion. Conclusion. Kahoot es una herramienta digital interactiva, gratuita y de manejo sencillo tanto para el docente como para los alumnos, que permite que estos mejoren en su aprendizaje haciendo uso de nuevas tecnologias y se sientan asi mas motivados. EnglishIntroduction. The need to face a new generation of students, the ‘Z generation’, considered as the first digital native generation, makes it necessary to implement new teaching methodologies in the university field to improve the teaching-learning process and above all achieve greater motivation in the student. To this end, the use of new information and communication technologies has enabled the dynamics of these processes and increasing student motivation at all educational levels, including university teaching. Subjects and methods. Use of the Kahoot educational app was raised through mobile devices as an innovative tool that enhances motivation and learning process, participating students enrolled in the subject of ‘Human Anatomy II (Esplacnology)’, second course of the Degree of Medicine in the University of Zaragoza. Results. The impact that the experience has caused students has been studied through qualitative assessment surveys, analyzing the learning outcomes obtained by students. The results achieved in terms of implementation of these new teaching-learning methodologies have been valued by students, in a very positive way both at the level of usefulness in learning teaching and at the level of motivation. Conclusion. Kahoot is an interactive digital tool, free and easy to use for both teachers and students, and which allows the latter to improve their learning using new technologies, feeling more motivated.

Hypertrophic cardiomyopathy is the most common monogenic heart disease. Its phenotypic expression is quite variable. In up to 60% of the cases, mutations are described in the genes coding for cardiac sarcomer proteins. Massive sequencing of deoxyribonucleic acid makes it possible to discover new genes responsible for the disease, but it has the disadvantage of discovering numerous variants of uncertain significance in these patients. The strategy used, especially when they do not segregate with the disease, is one of the challenges of genetics. Pathogenicity criteria may help to catalogue this variant. The genetic tests on the index case a diagnosis to be made, and the possibility of cascading to first degree relatives. The presence or not of a positive genotype in the relatives will determine the subsequent follow-up guidelines. The appearance of a positive genotype is a poor prognosis regardless of the type of mutation.

Background & Aims: Little is known about the prevalence of functional gastrointestinal disorders (FGIDs) in children from the Mediterranean area of Europe. We aimed to assess the prevalence of FGIDs in children and adolescents in this region. Methods: We collected data on 13,750 children (4–18 years old) enrolled in the Mediterranean–European Area Project, a school-based health study performed in Croatia, Greece, Israel, Italy, Jordan, Lebanon, Macedonia, Serbia, and Spain. Data were collected from March to June and in September of 2016. We analyzed data from 6602 students 4 to 10 years old (group A; mean age, 7.7 ± 1.9 y), and 7148 subjects 11 to 18 years old (group B; mean age, 13.8 ± 2.1 y). Children with FGIDs were identified based on answers to questionnaires on pediatric gastrointestinal symptoms, selected based on Rome III criteria. Results: In group A, the prevalence of FGIDs was 20.7%. The most frequent disorders were functional constipation (11.7%), irritable bowel syndrome (IBS, 4%), aerophagia (3.5%), and abdominal migraine (3.1%). The prevalence of abdominal migraine was significantly higher in girls than in boys (P =.007). In group B, the overall prevalence of FGIDs was 26.6%. The most frequent disorders were functional constipation (13.1%), abdominal migraine (7.8%), aerophagia (6.3%), and IBS (5.6%). In group B, FGIDs had a higher prevalence among girls than boys (P

Yeasts used in fermentation processes are subject to different stress conditions, including low pH, high initial substrate concentration, accumulation of toxic compounds (e.g. ethanol), and temperature fluctuations. The fermentation of mescal is conducted under variable environmental conditions throughout the year. One of the most important environmental factors is temperature, as autumn and winter fermentations occur at low temperatures, but fermentation temperatures are much higher in summertime. The aim of this work was to compare the response of Saccharomyces cerevisiae strains isolated from agave fermentations to different stress conditions and two different medium culture one mimicking mezcal production, using agave must and other synthetic must development for analysis in wine industry. The strains isolated from agave were compared with a commercial strain used in wine elaboration, which exhibits good tolerance to the different stresses found in industrial fermentations. All strains grew in the presence of glucose and fructose irrespective of the sugar concentration, and low pH did not affect the formation of colonies. Differences in growth were observed among the strains at low temperature and high concentrations of ethanol. Only 28% of the tested strains exhibited good tolerance to high ethanol concentrations, a desirable trait for avoiding stuck fermentations. The strain ITD00185 was able to grow in alcoholic stress condition, consuming sugar and producing ethanol in agave must. This strain shows tolerance to the different stress conditions and may be a useful starter culture for agave fermentation, the potential of the native strains to be used to improve other industrial fermentation processes that involve low temperatures and high ethanol yields. Key words: Ethanol, tolerance, Mescal, native strains.

We screened BRCA1 and BRCA2 germline mutations in 60 high-risk breast and/or ovarian cancer patients and 20 relatives from Aragon (Spain) by DHPLC (Denaturing High Performance Liquid Chromatography) and direct sequencing of the entire coding sequence and the splicing sites of both genes. We have identified 17 different pathogenic mutations: 8 in BRCA1 and 9 in BRCA2 in 60 unrelated patients and 50% of relatives were carriers. The prevalence of pathogenic mutations in this study was 33.33%. Two truncating mutations are novel: c.5024_5025delGA in exon 16 of BRCA1 and c.2929delC in exon 11 of BRCA2 (numbered after GenBank U14680 and U43746). Multiplex Ligation Dependent Probe Amplification (MLPA) was performed for large mutational scanning of both genes and a large genomic deletion in BRCA1 was found (DelEx8-13). Furthermore, five mutations are described for the first time in Spanish population: c.1191delC, c.3478_3479delTT and c.6633_6637delCTTAA (BRCA1) and c.3972_3975delTGAG and 3908_3909delTG (BRCA2). Three mutations have been reported previously once in Spain: c.3600_3610del11 (BRCA1), c.5804_5807delTTAA (BRCA2) and c.9246C>A (BRCA2). The mutation c.5374_5377delTATG has been found before only in two unrelated families from Castilla-Leon, Spain (BRCA2). Frequent mutations described in Spanish population have also been present: c.187_188delAG, c.5242C>A and c.5385insC in BRCA1 and c.3492_3493insT and c.9254_9258delATCAT in BRCA2. c.5242C>A, 3972_3975delTGAG and c.5804_5807delTTAA were the recurrent mutations found. Fifteen different unclassified variants were identified (25% families). Although specific BRCA1 and BRCA2 mutations are recurrently reported as a result of genetic founder effects we conclude that heterogeneous ethnicity increases the variety of mutations that can be found in Spanish populations.

Summary Background There is an overexpression of cyclo-oxygenase 2 (COX-2) in Barrett’s oesophagus (BO). Aim To determine the long-term effect of a COX-2 inhibitor on cellular mechanisms involved in BO. Methods A randomized controlled trial was conducted in BO patients allocated to continue the usual proton pump inhibitor (PPI) alone treatment, or PPI combined with rofecoxib (25 mg/day) for 6 months. Cell proliferation index and COX-2 expression in BO glands was determined in biopsy specimens at baseline and after treatment. Cell apoptosis, cyclin D1, p53 and vascular endothelial growth factor (VEGF) expression was also explored in a subset of patients. Student-t test and the U-Mann–Whitney test were used for quantitative and ordinal variables. Results Of 62 patients, 58 completed the study. A higher proportion of patients on rofecoxib + PPI exhibited a decrease in COX-2 expression compared to those treated with PPI alone, but cell proliferation index was not affected. Unlike PPI alone, rofecoxib + PPI was associated with an increase in the apoptotic cell index, a decrease in p53 cell staining and VEGF expression in mucosal vessels. No effect on low-grade dysplasia or cyclin D1 was observed. Conclusions The addition of rofecoxib to PPI therapy does not affect cell proliferation index in BO cells after 6 months of therapy, but does reduce COX-2 and VEGF expression and increases cell apoptosis.

Cytokine balance alterations are responsible for some of the systemic and hepatic manifestations of alcoholism. The present study was aimed to evaluate the influence of both acute alcohol abstinence (in alcoholics) and acute alcohol intake (in healthy subjects) on serum IL-6, IL-8, IL-10, and IL-12 levels. Serum cytokine concentrations were determined on admission and after a median of 6 days of ethanol abstinence in 29 patients with alcohol withdrawal syndrome. The same determinations were made in five healthy volunteers at baseline and after 36 h of a single 60 g-dose alcohol intake. Increased serum levels of IL-6, IL-10 and, to a lesser extent IL-8, declined in the few days after alcohol abstinence in patients with alcohol withdrawal syndrome. Serum IL-8 values increased after alcohol intake in healthy subjects. Rapid variation of serum cytokine levels along with alcohol intake or abstinence should be taken into account in cytokine studies in alcohol abusers.

Background: Fever is considered common during alcohol withdrawal, but there are very few reports focused on this problem. The aim of the present study was to evaluate the prevalence and causes of fever in patients admitted to the hospital with alcohol withdrawal syndrome. Methods: A total of 110 episodes of alcohol abstinence syndrome (10 cases of tremulousness and anxiety, 38 of delirium, and 62 cases of seizure) in 98 patients admitted to the hospital for that reason (82 males and 12 females, median age 42 years, range 18–72 years) were reviewed. Cases with fever on admission were not included. Results: Seventy-two cases (65%) presented low-grade fever (maximum axillary temperature 37–37.9°C), and 23 cases (21%) presented high-grade fever (≥38°C). Pyrexia appeared after a median of 2 days in the hospital (range 1–8 days). The prevalence of fever was similar among the different alcohol withdrawal syndromes. Catheter-associated phlebitis was the cause of pyrexia in 39 cases (41%). Respiratory infection was present in 15 cases of pyrexia (16%) and urinary tract infection in 11 (12%). Patients with low serum albumin levels had a higher rate of both respiratory and urinary tract infection. Ten patients (12%) had more than one cause of fever. In 38 cases of pyrexia (40%), no cause was identified. This was more frequent in low-grade fever (33 cases, 46%) than in high-grade fever (five cases, 22%). All cases of pyrexia without an obvious cause resolved spontaneously after a median of 3 days. Conclusions: Pyrexia is very common during alcohol withdrawal syndrome. A cause may frequently be identified, particularly in patients with high-grade fever. Many patients have transient pyrexia (mostly low-grade fever) without an obvious cause.

It has been reported that total serum IgE is increased in patients with alcoholic cirrhosis, but it is not clear if this fact is related to alcoholic liver disease or to alcohol intake.To measure serum IgE in a group of chronic alcoholics with different stages of liver injury in order to elucidate if IgE increase in related to alcoholic liver damage.Total serum IgE was determined by enzyme immunoassay in 186 chronic alcoholic patients (137 male/49 female) and 101 healthy controls. Patients and controls with known reasons for IgE elevation were excluded. Among alcoholic patients, 24 had fatty liver, 28 hepatic fibrosis, 29 alcoholic hepatitis, and 67 liver cirrhosis (38 patients were not evaluable concerning liver injury).Total serum IgE was found to be increased in alcoholics (median 154.5 IU/mL, range 1-7329 IU/mL) with respect to healthy controls (median 20 IU/mL, range1-1417 IU/mL) (P0.001). IgE increase was moderate (180-1000 IU/mL) in 60 alcoholics (32.3%) and marked (1000 IU/mL) in 27 (14.5%). Male alcoholics had higher IgE levels than females (median 191 IU/mL and range 1-7329 IU/mL vs 105 IU/mL and range 2-2189 IU/mL) ( P = 0.009). On logistic regression analysis, alcoholism, male sex and younger age (but not smoking) were independently associated with higher IgE levels. No clear relationship was noted between serum IgE and severity of alcoholic liver disease. Thus, no correlation was observed between IgE and parameters of liver function (serum bilirubin, albumin or prothrombin index). Likewise, IgE concentrations were not significantly different in patients with liver cirrhosis with respect to patients with less severe liver disease. Serum IgE was increased (180 IU/mL) in 47.8% of cirrhotics and in 44% of patients without liver cirrhosis. In contrast, other immunoglobulins (IgG, IgA and IgM) were significantly correlated with liver dysfunction.Chronic alcoholism should be considered as a cause of increased total serum IgE, regardless of the severity of the underlying liver disease.

The immunoglobulin G binding site in the globular regions of human complement subcomponent C1q has been investigated by chemical modification of histidine residues with diethylpyrocarbonate and arginine residues with phenylglyoxal and cyclohexane-1,2-dione (CHD). Only the modification of arginine residues with CHD fulfills the requirements of a specific modification without unwanted side reactions. Specific modification of arginine residues with CHD results in loss of immune complex recognition without affecting the binding of C1r2S2 to form C1. The gross structure of C1q is not changed by CHD treatment, and immune complex binding is restored to 82% of the control upon NH2OH treatment. Enzymic digestion and isolation of the modified peptides indicate that the modification by CHD of 4 to 5 arginine residues (A162, B114, B129, C156, and possibly B163) per C1q globular "head" abolishes the ability of C1q to interact with immune complexes. These residues define two areas (and possible binding sites for IgG) on the globular region of C1q: B114-B129 (site 1) and A162-(B163)-C156 (site 2). Sequence comparison and solvent exposure predictive studies favor site 2 as the immunoglobulin G binding site on the globular regions of C1q, although the participation of site 1 cannot be ruled out.

An experimental rat liver cirrhosis, by means of carbon tetrachloride and ethanol during 8 weeks, was employed to assay the effect of Nifedipine (a calcium antagonist blocker), S-Adenosylmethionine (a precursor of glutathione); singly and in combination on rat liver cirrhosis. A slight decrease of cirrhosis (N.S.) was observed with the pharmacological therapy employed singly. The combination therapy (Nifedipine+S-Adenosylmethionine) significantly inhibited liver cirrhosis (p < 0.01).

Genetic relationships among forty-one strains of Saccharomyces bayanus var. uvarum isolated in different wine regions of Europe and four wild isolates were investigated by restriction analysis (RFLP) of mitochondrial DNA (mtDNA) with four restriction endonucleases, AluI, DdeI, HinfI and RsaI. No clear correlation between origin and source of isolation of S. bayanus var. uvarum strains and their mtDNA restriction profiles was found. On the whole, the mtDNA of S. bayanus var. uvarum is much less polymorphic than that of S. cerevisiae. This observation is in good agreement with results obtained by electrophoretic karyotyping. Unlike wine S cerevisiae, strains of S. bayanus var. uvarum display a low level of chromosome length polymorphism.

An experimental model of toxic liver injury in rats was employed to assay the effect of Nifedipine (a calcium antagonist blocker) and S-Adenosylmethionine (a precursor of glutathione). An important decrease in both perivenular fibrosis and cirrhosis was found. Furthermore, a significant decrease in lactic acid levels was found in the group of animals treated with pharmacologic therapy, although no correlation was seen between lactic acid levels and the different degrees of perivenular fibrosis. No significant variations in ALT and AST enzymes were observed between both groups, as opposed to a significant decrease in LDH enzyme in the Nifedipine+S-Adenosylmethionine group. The results indicate an improvement in the histologic picture of the liver in rats treated by means of pharmacological association, without any change in inflammatory infiltrate and with a slight decrease in necrosis, indicating an action mechanism via creeping fibrosis (instead of a hepatitis pathway).

There is an overexpression of cyclo-oxygenase 2 (COX-2) in Barrett's oesophagus (BO).To determine the long-term effect of a COX-2 inhibitor on cellular mechanisms involved in BO.A randomized controlled trial was conducted in BO patients allocated to continue the usual proton pump inhibitor (PPI) alone treatment, or PPI combined with rofecoxib (25 mg/day) for 6 months. Cell proliferation index and COX-2 expression in BO glands was determined in biopsy specimens at baseline and after treatment. Cell apoptosis, cyclin D1, p53 and vascular endothelial growth factor (VEGF) expression was also explored in a subset of patients. Student-t test and the U-Mann-Whitney test were used for quantitative and ordinal variables.Of 62 patients, 58 completed the study. A higher proportion of patients on rofecoxib + PPI exhibited a decrease in COX-2 expression compared to those treated with PPI alone, but cell proliferation index was not affected. Unlike PPI alone, rofecoxib + PPI was associated with an increase in the apoptotic cell index, a decrease in p53 cell staining and VEGF expression in mucosal vessels. No effect on low-grade dysplasia or cyclin D1 was observed.The addition of rofecoxib to PPI therapy does not affect cell proliferation index in BO cells after 6 months of therapy, but does reduce COX-2 and VEGF expression and increases cell apoptosis.

The fermentation of lactose (Lac+) in the dairy yeast Kluyveromyces lactis var. lactis is controlled by the LAC4 (beta-galactosidase) and LAC12 (lactose permease) genes. The complementation analysis of twelve Kl. lactis var. drosophilarum natural homothallic Lac- strains of different origin was carried out using the genetic heterothallic lines of Kl. lactis var. lactis of the lac4LAC12 and LAC4lac12 genotypes. It was shown that the natural Lac- strains did not possess the LAC4LAC12 gene cluster. Southern hybridization of chromosomal DNA with LAC4 and LAC12 probes, as well as recombination analysis, showed that Kl. lactis var. drosophilarum yeasts do not have even silent copies of these genes. As distinct from this yeast, natural Lac- strains of the yeast Kl. marxianus are mutants impaired in the lactose permease gene (lac12 analogue), but possess an active beta-galactosidase gene (LAC4 analogue). The origin of the LAC4LAC12 gene cluster of the dairy yeasts Kl. lactis is discussed.

Many symbioses between bacteria and insects resulted from ancient infections followed by strict vertical transmission within host lineages. The strong bottlenecks under which this transmission occurs promote the neutral fixation of slightly deleterious mutations by genetic drift. As predicted by Muller's ratchet, this fixation will drive endosymbiotic bacteria through an irreversible dynamics of fitness loss. The chaperonin GroEL has been proposed as a compensatory mechanism whereby endosymbiotic bacteria of aphids persist. Here, we show that endosymbiotic bacteria of insects from two phylogenetically very distant bacterial phyla have fixed amino acid substitutions by positive selection in functionally important GroEL regions involved in either GroES/peptide binding or in the en bloc movement of the GroEL apical domain. These results, together with the high levels of constitutive expression of GroEL in these endosymbionts, provide valuable insights into the evolution of a molecular mechanism responsible for the maintenance of the symbiotic lifestyle.

We studied the rate of apoptosis in the placental tissue of pregnancies complicated with intrauterine growth retardation (IUGR) and compared it with the results obtained in normal placentas. Our results clearly demonstrate a strongly increased rate of apoptosis in placentas of children born with IUGR, suggesting severe placental dysfunction. The significance of these findings needs further study.

To investigate the possibility of altered gene expression of growth factors in prenatal growth retardation, we assessed expression of the genes for insulin-like growth factor-I (IGF-I) and epidermal growth factor receptor (EGFR) by RT-PCR from human placentas at term delivery in two groups: appropriate for gestational age (AGA) and pregnancies complicated with IUGR. The placentas from IUGR gestations showed reduced IGF-I expression with a significance of p = 0.008, whereas we did not find any significant differences in EGFR gene expression.

Background: Withdrawal syndrome is a hallmark of alcohol dependence. The characteristics of alcohol consumption, closely related to dependence, could influence the development of alcoholic liver disease. The study aimed to investigate if patients with severe alcohol withdrawal syndrome have a peculiar profile of liver disease. Methods: The study included 256 heavy drinkers (aged 19–75 years, 70.3% males) admitted to an Internal Medicine Department. Patients admitted for complications of liver disease were not included. Severe alcohol withdrawal syndrome (seizures, disordered perceptions, or delirium) developed in 150 patients (58.6%). Alcohol consumption (daily quantity, duration, and pattern [regular or irregular]) was assessed by questionnaire. Liver biopsy was performed in all cases. Results: Patients with alcohol withdrawal syndrome showed a lower prevalence of liver cirrhosis and a higher prevalence of alcoholic hepatitis than patients without it. The negative association of alcohol withdrawal syndrome with liver cirrhosis persisted after we adjusted for sex, daily intake, duration, and pattern of alcohol consumption. Alcoholic hepatitis was independently associated with the irregular pattern of alcohol consumption, which was closely associated with severe alcohol withdrawal syndrome. Conclusions: The profile of liver injury is different in heavy drinkers who develop and who do not develop a severe alcohol withdrawal syndrome when admitted to the hospital.

HBV, HCV, and HIV have some transmission routes in common. Viral liver disease is a leading cause of mortality in HIV-infected patients. The study was aimed at evaluating the prevalence of HBV and HCV markers in subjects with different risk practices for HIV infection.A total of 699 subjects were studied Of these subjects, 517 were intravenous drug users (373 HIV-positive and 144 HIV-negative), 127 had heterosexual risk practice (66 HIV-positive and 61 HIV-negative), 31 had homosexual risk practice (all HIV-positive), 15 had post-transfusional HIV infection, and nine had HIV infection of unknown source. Patients with anti-HBc antibody were considered HBV-positive, and cases with anti-HCV antibodies were considered HCV-positive.Among patients with HIV infection, most intravenous drug users (79%) had markers of both HBV and HCV, compared with 20%, 11%, and 10% of cases infected by transfusional, heterosexual, and homosexual route, respectively (p0.001). Absence of both HBV and HCV markers was observed in most HIV-positive heterosexuals (62%) compared with 40% of post-transfusional cases, 32% of homosexuals and 4% of intravenous drug users (p: NS, p = 0.009, and p0.001, respectively). Isolated HBV-positivity was the most frequent pattern in HIV-infected homosexuals (58%), compared with 27% of post-transfusional, 21% of heterosexuals and 11% of intravenous drug users (p: NS, p0.001 and p0.001, respectively). HIV-negative intravenous drug users had a lower prevalence of HBV/HCV association than HIV-positive cases (p0.001). Isolated HCV-positivity was more frequent in HIV-negative than in HIV-positive intravenous drug users (27% vs. 6%, p0.001). In heterosexuals, isolated HBV-positivity was more prevalent in HIV-positive than in HIV-negative cases (21% vs. 7%, p = 0.04).HBV and HCV seroprevalence in HIV infected patients vary depending on the risk practice. This suggests a variable transmissibility depending on the route considered. Within the same risk practice, differences in HCV and HBV seroprevalence between HIV-positive and HIV-negative cases suggest that some factors associated with HIV infection may influence the rate of infection by HCV and HBV.

To determine the clinical manifestations of the relapse of the tuberculosis (TB) in the patients infected by the human immunodeficiency virus (HIV) and the degree of immunodeficiency shown.They are analyzed retrospectively 121 cases, all they HIV+, diagnosed of TB from November of 1985 until December of 1999. All they fulfilled correctly the specific treatment of the tuberculosis.Of 121 studied cases, a total of 13 (10.7%) presented relapse of tuberculosis finally of a mean of 31.8 months (range 1-95). In 7 cases the affectation was pulmonary (2 typical, 5 non typical), in 3 mixed (pulmonary and extrapulmonary) and in 3 extrapulmonary. The clinical sintomatology were present in 92% of the cases and, predominantly, was: fever (92%), cough (69%), sweating, weight loss (61%). Hepatomegaly results be the exploratory finding quite frequent (54%). The mean of CD4+ lymphocytes was 141.7 (range 3-406), in 8 (61%) the lymphocyte counts was inferior to 100 cells/mm3. PPD was positive in 1 case (14%) and with respect to delayed-type hypersensitivity reaction test, it was observed not normoergia in all the cases (100%).Relapse of the tuberculosis in patient infected by the VIH is presented with a manifested clinical expresivity in the course of a severe degree of immunodeficiency. In certain groups of risk as the intravenous drugs users, it must of be had a high level of suspicion of the relapse of the disease, above all in those patients that show low CD4+ lymphocyte counts.

Objetivo: Determinar las manifestaciones clínicas de la recidiva de la tuberculosis (TB) en los pacientes infectados por el virus de la inmunodeficiencia humana (VIH) y el grado de inmunodeficiencia mostrado. Material y métodos: Se analizan retrospectivamente 121 casos, todos ellos VIH+, diagnosticados de TB desde noviembre de 1985 hasta diciembre de 1999. Todos ellos habían cumplido correctamente el tratamiento específico de la tuberculosis. Resultados: De los 121 casos estudiados, un total de 13 (10,7%) presentaron recidiva de tuberculosis al cabo de una media de 31,8 meses (rango 1-95). En 7 casos la afectación era pulmonar (2 típica, 5 atípica), en 3 mixta (pulmonar y extrapulmonar) y en 3 extrapulmonar. La sintomatología estuvo presente en el 92% de los casos y, predominantemente, consistió en: fiebre (92%), tos (69%), sudoración, adelgazamiento (61%). La hepatomegalia fue el hallazgo exploratorio más frecuente (54%). La media de linfocitos CD4 fue de 141.7 (rango 3-406), en 8 (61%) los recuentos era inferiores a 100 cels/mm³. La PPD fue positiva en 1caso (14%) y en la prueba de hipersensibilidad retardada, se observó no normoergia en todos los casos (100%). Conclusiones: La recidiva de la tuberculosis en pacientes infectados por el VIH se presenta con una manifiesta expresividad clínica en el ámbito de un grado severo de inmunodeficiencia. En ciertos grupos de riesgo como los usuarios de drogas por vía parenteral, debe de tenerse un alto índice de sospecha de la recidiva de la enfermedad, sobre todo en aquellos pacientes que muestren recuentos celulares bajos de linfocitos CD4.

Phylogenetic reconstruction methods are subject to two types of limitations: our knowledge about the true history of organisms and the gross simplification implied in the numerical simulation models of the relationships between them. In such a situation, experimental phylogenetics provides a way to assess the accuracy of the phylogenetic reconstruction methods. Nonetheless, this capacity is only feasible for organisms in which replication and mutation rates are high enough to provide valuable data. On the other hand, experimental phylogenetics also provides insights on the main evolutionary processes acting on viral variability under different population dynamics. Our study with the foot-and-mouth disease virus (FMDV) strongly suggests that the phylogenetic reconstruction methods can infer erroneous phylogenies due to nucleotide convergences between isolates belonging to different experimental lineages. We also point out that the diverse evolutionary mechanisms acting in different experimental dynamics generate alterations and change the frequencies of genetic variants, which can lead to the misinterpretation of the real evolutionary history.

The PCR amplification and subsequent restriction analysis of the region spanning the internal transcribed spacers (ITS1 and ITS2) and the 5.8S rRNA gene was applied to the identification of yeasts belonging to the genus Saccharomyces. This methodology has previously been used for the identification of some species of this genus, but in the present work, this application was extended to the identification of new accepted Saccharomyces species (S. kunashirensis, S. martiniae, S. rosinii, S. spencerorum, and S. transvaalensis), as well as to the differentiation of an interesting group of Saccharomyces cerevisiae strains, known as flor yeasts, which are responsible for ageing sherry wine. Among the species of the Saccharomyces sensu lato complex, the high diversity observed, either in the length of the amplified region (ranged between 700 and 875 bp) or in their restriction patterns allows the unequivocal identification of these species. With respect to the four sibling species of the Saccharomyces sensu stricto complex, only two of them, S. bayanus and S. pastorianus, cannot be differentiated according to their restriction patterns, which is in accordance with the hybrid origin (S. bayanus x S. cerevisiae) of S. pastorianus. The flor S. cerevisiae strains exhibited restriction patterns different from those typical of the species S. cerevisiae. These differences can easily be used to differentiate this interesting group of strains. We demonstrate that the specific patterns exhibited by flor yeasts are due to the presence of a 24-bp deletion located in the ITS 1 region and that this could have originated as a consequence of a slipped-strand mispairing during replication or be due to an unequal crossing-over. A subsequent restriction analysis of this region from more than 150 flor strains indicated that this deletion is fixed in flor yeast populations.

To analyse the clinical manifestations and performance of the diagnosis methods in tuberculosis (TB) diagnosed in patients with human immunodeficiency virus infection (HIV), according to the location forms of disease (LF).We revised 80 cases of TB diagnosed in patients with infection by HIV. The data were gathered in relation to TB location, the clinical data and the microbiological and histological diagnosed studies. In the statistic analysis the values p0.05 were taken into account.The prevalence of LF was: 1) pulmonary forms (PF) 32 (40%), of which 12 were typical pulmonary (TP) and 20 atypical pulmonary (AP); 2) mixed forms (MF) 21 (26%); 3) extrapulmonary forms (EF) 19 (24%), of which 17 were lymphadenitis; 4) miliary tuberculosis (MT) 8 cases (10%). The prevailing symptoms were: fever (71%) mainly in MF and MT, cough (69%), with less frequency in AP, EF and MT (p0.05) and adenomegalies especially in EF and MF. The diagnosis was based on the study of the sputum smears and lymph node. The bacilloscope of the spontaneous sputum was 53% with a minor performance in the AP (p0.05), while in lymph node the smear for AFB was positive in 78% cases and caseous granulomas were observed in 87%.In our study the major confirmed locations were pulmonary and lymph node, the most important clinical symptoms were fever, cough and adenomegalies. The diagnosis was based, in the most of cases, on the microbiologic and histologic examination of bronchial secretion and lymph node samples.

Objetivo: El objeto de este trabajo fue determinar la inmunidad celular utilizando el Multitest CMI y correlacionar su resultado con el recuento de linfocitos CD4, y el riesgo de desarrollar tuberculosis activa en el caso de que la pápula correspondiente a la tuberculosis del multitest fuera positiva. Material y Métodos: Se realizó un seguimiento longitudinal prospectivo de 342 pacientes UDVP, 210 infectados con el VIH-1 y 132 de ellos seronegativos. La cohorte objeto del estudio estaba formada por 165 UDVP seropositivos para el VIH-1 (128 varones y 37 mujeres). Las edades estaban comprendidas entre 18 y 45 años. El tiempo del trabajo fue de 25 meses (8-25). La prueba de la tuberculina y el multitest les fueron practicados al inicio y cada tres meses así como un despistaje de tuberculosis activa a lo largo de todo el periodo del estudio cuando se consideró necesario. Resultados: Se desarrolló tuberculosis activa en 9,25 casos por cien personas y año de seguimiento en los estados con mejor nivel inmunológico (score de la PHR >10 mm). Había una correlación significativa entre el diámetro de la pápula de la prueba de la tuberculina y la pápula de la fracción tuberculínica de la prueba de hipersensibilidad retardada. Conclusiones: La prueba de la hipersensibilidad retardada realizada con el Multitest IMC es de utilidad para evaluar la función de la inmunidad celular y en los pacientes seropositivos para el VIH-1 anérgicos o con la fracción tuberculínica de la PHR positiva debe considerarse la posibilidad de quimioprofilaxis con isoniazida (9 meses).

A retrospective study of 80 HIV patients diagnosed with tuberculosis was carried out in order to evaluate the clinical presentation of tuberculosis (CPT) in relation to the degree of HIV-induced immunodeficiency, as determined by the CD4 lymphocyte count and reactivity to the tuberculin and delayed-type hypersensitivity reaction (DHR) skin tests by applying 2TU of RT-23 tuberculin, and the Muiltest IMC Merieux, respectively. CPT classification was undertaken on the basis of the location of the disease: pulmonary tuberculosis (PT), distinguishing between typical pulmonary (TP) and atypical (AP) according to the radiological pattern; extrapulmonary (ET); mixed tuberculosis (MT) pulmonary and extrapulmonary; and miliary tuberculosis. The CD4 lymphocyte count was 264.6 +/- 226.8, the TP had the highest number (505), MT had 132 (p < 0.001) and the miliary tuberculosis had 148 (p < 0.001), the lowest. The tuberculin skin test was positive in 35%, of which 11% were MT (p < 0.05). In the delayed-type hypersensitivity reaction, 67% were non-normoergic, of which 95% and 100% were MT and miliary tuberculosis, respectively (p < 0.05). There was a good overlap between CD4 depletion and skin tests. TP exhibited moderate immunodeficiency, AP severe immunodeficiency, and mixed and miliary TB extremely high immunodeficiency.