Clues on the dynamics of DNA replication in Giardia lamblia Icon for the Forest of Biologists

Genomic replication is a critical, regulated process that ensures accurate genetic information duplication. In eukaryotic cells, strategies have evolved to prevent conflicts between replication and transcription. Giardia lamblia, a binucleated protozoan, alternates between tetraploid and octoploid genomes during its cell cycle. Using single-molecule techniques like DNA combing and nanopore-based sequencing, we investigated the spatio-temporal organization of DNA replication, replication fork progression, and potential head-on replication-transcription collisions in Giardia trophozoites. Our findings indicate that Giardia chromosomes are replicated from few active origins, which are widely spaced and exhibit faster replication rates compared to other protozoan parasites. Immunofluorescence assays revealed that around 20% of trophozoites show asynchronous replication between nuclei. Forksense and gene ontology analyses disclosed that genes in regions with potential head-on collisions are linked to chromatin dynamics, cell cycle regulation, and DNA replication/repair pathways, possibly explaining the observed asynchronous replication in part of the population. This study offers the first comprehensive view of replication dynamics in Giardia, the cause of giardiasis, a diarrheal disease impacting millions worldwide.

Immunodiagnosis of cystic echinococcosis in livestock: Development and validation dataset of an ELISA test using a recombinant B8/2 subunit of Echinococcus granulosus sensu lato.

The accuracy of screening tests for detecting cystic echinococcosis (CE) in livestock depends on characteristics of the host-parasite interaction and the extent of serological cross-reactivity with other taeniid species. The AgB8 kDa protein is considered to be the most specific native or recombinant antigen for immunodiagnosis of ovine CE. A particular DNA fragment coding for rAgB8/2 was identified, that provides evidence of specific reaction in the serodiagnosis of metacestode infection. We developed and validated an IgG Enzyme Linked Immunosorbent Assay (ELISA) test using a recombinant antigen B sub-unit EgAgB8/2 (rAgB8/2) of Echinoccocus granulosus sensu lato (s.l.) to estimate CE prevalence in sheep. A 273 bp DNA fragment coding for rAgB8/2 was expressed as a fusion protein (∼30 kDa) and purified by affinity chromatography. Evaluation of the analytical and diagnostic performance of the ELISA followed the World Organisation for Animal Health (OIE) manual, including implementation of serum panels from: uninfected lambs (n = 79); experimentally infected (with 2,000 E. granulosus s.l. eggs each) sheep with subsequent evidence of E. granulosus cysts by necropsy (n = 36), and animals carrying other metacestode/trematode infections (n = 20). The latter were used to assess the cross-reactivity of rAgB8/2, with these animals being naturally infected with Taenia hydatigena, Thysanosoma actinioides and/or Fasciola hepatica. EgAgB8/2 showed cross-reaction with only one serum sample from a sheep infected with Ta. hydatigena out of the 20 animals tested. Furthermore, the kinetics of the humoral response over time in five 6-month old sheep, each experimentally infected with 2,000 E. granulosus s.l. eggs, was evaluated up to 49 weeks (approximately one year) post infection (n = 5). The earliest detectable IgG response against rAgB8/2 was observed in sera from two and four sheep, 7 and 14 days after experimental infection, respectively. The highest immune response across all five animals was found 16 to 24 weeks post infection.

Modelling diagnostics for Echinococcus granulosus surveillance in sheep using Latent Class Analysis: Argentina as a case study.

Echinococcus granulosus sensu lato is a globally prevalent zoonotic parasitic cestode leading to cystic echinococcosis (CE) in both humans and sheep with both medical and financial impacts, whose reduction requires the application of a One Health approach to its control. Regarding the animal health component of this approach, lack of accurate and practical diagnostics in livestock impedes the assessment of disease burden and the implementation and evaluation of control strategies. We use of a Bayesian Latent Class Analysis (LCA) model to estimate ovine CE prevalence in sheep samples from the Río Negro province of Argentina accounting for uncertainty in the diagnostics. We use model outputs to evaluate the performance of a novel recombinant B8/2 antigen B subunit (rEgAgB8/2) indirect enzyme-linked immunosorbent assay (ELISA) for detecting E. granulosus in sheep. Necropsy (as a partial gold standard), western blot (WB) and ELISA diagnostic data were collected from 79 sheep within two Río Negro slaughterhouses, and used to estimate individual infection status (assigned as a latent variable within the model). Using the model outputs, the performance of the novel ELISA at both individual and flock levels was evaluated, respectively, using a receiver operating characteristic (ROC) curve, and simulating a range of sample sizes and prevalence levels within hypothetical flocks. The estimated (mean) prevalence of ovine CE was 27.5% (95%Bayesian credible interval (95%BCI): 13.8%-58.9%) within the sample population. At the individual level, the ELISA had a mean sensitivity and specificity of 55% (95%BCI: 46%-68%) and 68% (95%BCI: 63%-92%), respectively, at an optimal optical density (OD) threshold of 0.378. At the flock level, the ELISA had an 80% probability of correctly classifying infection at an optimal cut-off threshold of 0.496. These results suggest that the novel ELISA could play a useful role as a flock-level diagnostic for CE surveillance in the region, supplementing surveillance activities in the human population and thus strengthening a One Health approach. Importantly, selection of ELISA cut-off threshold values must be tailored according to the epidemiological situation.

Gastroprotective activity of kaempferol glycosides from Malvaviscus arboreus Cav.

ETHNOPHARMACOLOGICAL RELEVANCE: Malvaviscus arboreus is traditionally used in Mexico and Central America for culinary and medicinal purposes. Leaves and flowers of this species are commonly used for preparation of salads, herbal teas and herbal dyes. Panamanian, Guatemalan and Mexican healers use this medicinal plant for the management of fever, respiratory complications, dysentery, liver and gallbladder problems, stomachache and gastritis between other health troubles. AIM OF THE STUDY: Considering the traditional use of M. arboreous as well as its content in flavonoids and other polyphenols, the objective of this work was to evaluate the gastroprotective effect of an aqueous extract and identify the potential bio-active principles from flowers of this species. MATERIAL AND METHODS: Fresh flowers of Malvaviscus arboreus were collected, dried, and macerated with water. The aqueous extract (ExAq) was partitioned using an immiscible mixture of water and ethyl acetate, giving an aqueous (MaAq) and organic (MaEA) fractions. The gastroprotective effect was carried out using an ethanol-induced gastric ulcer experimental test in male rats. While tween 20 was used as a negative control, famotidine (10 mg/kg) and L-arginine (300 mg/kg) were used as positive controls. Compounds 1 and 2 were isolated by several chromatographic techniques and the chemical characterization was carried out by means of the analysis of the NMR spectra in one and two dimensions. RESULTS: The integrate extract (ExAq) to 250, 500 and 750 mg/kg showed gastroprotective effect with high levels of 97.8%, 79.5% and 91.1% respectively. The organic fraction (MaEA) displayed a protection of 91.2%, 96.0% and 99.4% when it was evaluated at 125, 250 and 500 mg/kg respectively. Comparison of these results with famotidine at 10 mg/kg (83% of gastroprotection) indicated that ethyl acetate fraction showed a better gastroprotection. The bio-guided separation of this organic mixture, allowed obtaining the most active fraction (C1F4, 60 mg/kg) which was finally purified to obtain two glycosylated flavonols: kaempferol 3-O-D-sophoroside (1) and kaempferol 3-O-D-sambubioside (2). This mixture of flavonoids (40 y 60 mg/kg) showed 93.7 and 92% of gastroprotective activity respectively. CONCLUSION: This study allowed demonstrating that an aqueous extract and its organic fraction (MaEA) from M. arboreous contain glycosylated flavonoids (1 and 2) which are responsible of the gastroprotective properties of M. arboreous. These results will be used in the future development of a standardized treatment useful in the therapeutic management of gastric ulcers.

Active fractions of mannoproteins derived from yeast cell wall stimulate innate and acquired immunity of adult and elderly dogs.

Nutritional intervention in older dogs aims to increase lifespan and improve life quality as well as delay the development of diseases related to ageing. It is believed that active fractions of mannoproteins (AFMs) obtained through extraction and fractionation of yeast cell walls (Saccharomyces cerevisiae) may beneficially modulate the immune system. However, studies that have evaluated this component and the effects of ageing on the immune system of dogs are scarce. This study aimed to evaluate the immunological effects of AFMs in adult and elderly dogs. Three extruded iso-nutrient experimental diets were formulated: without addition of AFM (T0); with AFM at 400 mg/kg (T400); and with AFM at 800 mg/kg (T800). Thirty-six beagle dogs were used, and six experimental treatments, resulting in combinations of age (adult and elderly) and diet (T0, T400, and T800), were evaluated. On days zero, 14, and 28, blood samples were obtained for leucocyte phenotyping and phagocytosis assays. On days zero and 28, a lymphoproliferation test, quantification of reactive oxygen (H2O2) and nitrogen (NO) intermediate production, evaluation of faecal immunoglobulin A (IgA) content, and a delayed cutaneous hypersensitivity test (DCHT) were performed. Statistical analyses were performed with SAS software. Repeated measure variance analyses were performed, and means were compared by the Tukey test. Values of P ≤ 0.05 were considered significant, and values of P ≤ 0.10 were considered tendencies. Dogs fed T400 tended to have higher neutrophilic phagocytic activity than dogs fed T800 (P = 0.073). Regarding reactive oxygen intermediates, bacterial lipopolysaccharide (LPS)-stimulated neutrophils from animals that were fed T400 had a tendency to produce more H2O2 than those from animals fed the control diet (P = 0.093). Elderly dogs, when compared to adult dogs, had lower absolute T and B lymphocyte counts, lower auxiliary T lymphocyte counts, and higher cytotoxic T lymphocyte counts (P < 0.05). A significant effect of diet, age, and time with saline inoculation was noted for the DCHT. There was no effect of diet or age on faecal IgA content in dogs. This study suggests beneficial effects of mannoproteins on the specific and nonspecific immune responses in adult and elderly dogs.

XANTUS-EL: A real-world, prospective, observational study of patients treated with rivaroxaban for stroke prevention in atrial fibrillation in Eastern Europe, Middle East, Africa and Latin America.

BACKGROUND: The prospective, observational XANTUS study demonstrated low rates of stroke and major bleeding in real-world rivaroxaban-treated patients with non-valvular atrial fibrillation (NVAF) from Western Europe, Canada and Israel. XANTUS-EL is a component of the overall XANTUS programme and enrolled patients with NVAF treated with rivaroxaban from Eastern Europe, the Middle East and Africa (EEMEA) and Latin America. METHODS: Patients with NVAF starting rivaroxaban for stroke prevention were consecutively recruited and followed for 1 year, at approximately 3-month intervals, or for ≥30 days after permanent rivaroxaban discontinuation. Primary outcomes were major bleeding, adverse events (AEs), serious AEs and all-cause mortality. Secondary outcomes included stroke, non-central nervous system systemic embolism (non-CNS SE), transient ischaemic attack (TIA), myocardial infarction (MI) and non-major bleeding. All major outcomes were centrally adjudicated. RESULTS: Overall, 2064 patients were enrolled; mean age ±â€¯standard deviation was 67.1 ±â€¯11.32 years; 49.3% were male. Co-morbidities included heart failure (30.9%), hypertension (84.2%), diabetes mellitus (26.5%), prior stroke/non-CNS SE/TIA (16.2%) and prior MI (10.7%). Mean CHADS2, CHA2DS2-VASc and HAS-BLED scores were 2.0, 3.6 and 1.6, respectively. Treatment-emergent event rates were (events/100 patient-years, [95% confidence interval]): major bleeding 0.9 (0.5-1.4); all-cause mortality 1.7 (1.2-2.4); stroke/non-CNS SE 0.7 (0.4-1.2); any AE 18.1 (16.2-20.1) and any serious AE 8.3 (7.0-9.7). One-year treatment persistence was 81.9%. CONCLUSIONS: XANTUS-EL confirmed low stroke and major bleeding rates in patients with NVAF from EEMEA and Latin America. The population was younger but with more heart failure and hypertension than XANTUS; stroke/SE rate was similar but major bleeding lower.

Obtention and characterization of the recombinant simian Interleukin-15 in Escherichia coli for the preclinical assessment of an IL-15-based therapeutic vaccine.

Recombinant simian IL-15 (siIL-15) was obtained for the preclinical assessment of an anti-human IL-15 vaccine. For this purpose, the cDNA from peripheral blood mononuclear cells of a Macaca fascicularis monkey was cloned into a pIL-2 vector. The siIL-15 was expressed in Escherichia coli strain W3110 as an insoluble protein which accounted for 13% of the total cellular proteins. Inclusion bodies were solubilized in an 8 M urea solution, which was purified by ion exchange and reverse phase chromatography up to 92% purity. The protein identity was validated by electrospray ionization-mass spectrometry, confirming the presence of the amino acids which distinguish the siIL-15 from human IL-15. The purified siIL-15 stimulates the proliferation of cytotoxic T-lymphocytes line (CTLL)-2 and Kit 225 cells with EC50 values of 3.1 and 32.5 ng/mL, respectively. Antisera from modified human IL-15-immunized macaques were reactive to human and simian IL-15 in enzyme-linked immunosorbent assays. Moreover, the anti-human IL-15 antibodies from immune sera inhibited siIL-15 activity in CTLL-2 and Kit 225 cells, supporting the activity and purity of recombinant siIL-15. These results indicate that the recombinant siIL-15 is biologically active in two IL-15-dependent cell lines, and it is also suitable for the preclinical evaluation of an IL-15-based therapeutic vaccine.

Abuso sexual na infância e suas repercussões na vida adulta / Abuso sexual en la infancia y sus repercusiones en la vida adulta / Sexual abuse in childhood and its repercussions in adult life

RESUMO Objetivo: compreender repercussões do abuso sexual na vida adulta de mulheres abusadas sexualmente na infância. Método: pesquisa qualitativa, desenvolvida com nove mulheres em um Centro de Referência da Mulher, na região do semiárido do Estado de Pernambuco, Brasil, através de entrevistas não estruturadas. A interpretação dos resultados foi fundamentada na abordagem da Sociologia Compreensiva e do Cotidiano, a fim de identificar a centralidade subterrânea submersa na vida cotidiana das participantes e que pudesse emergir por meio de suas reações a partir do vivido do abuso sexual. Resultados: após a interpretação, os resultados foram agrupados nas categorias: Convivência familiar após a revelação do abuso sexual; A vida cotidiana de meninas em vivência de abuso sexual; e Repercussões do abuso sexual na vida adulta de mulheres abusadas sexualmente na infância. As repercussões do abuso sexual foram descritas como dificuldades no convívio familiar, gravidez, conduta hipersexualizada, prostituição, contradição entre gênero e sexo, dificuldades para ter orgasmo, uso de drogas, baixa autoestima, depressão, comportamento autodestrutivo, ideias suicidas e homicidas. Conclusão: a exposição ao abuso sexual no contexto familiar prejudicou a saúde física e emocional de meninas e adolescentes, bem como a convivência familiar, apontando para a necessidade de adoção de sensibilidade e solidariedade no cuidado a mulheres com queixas que possam estar associadas a vivências de abuso sexual.

RESUMEN Objetivo: comprender las repercusiones del abuso sexual en la vida adulta de mujeres abusadas sexualmente en la infancia. Método: investigación cualitativa, desarrollada con nueve mujeres en un Centro de Referencia de la Mujer, en la región del semi-árido del Estado de Pernambuco, Brasil, a través de entrevistas no estructuradas. La interpretación de los resultados fue fundamentada en el abordaje de la Sociología Comprensiva y del Cotidiano, a fin de identificar la centralidad subterránea inmersa en la vida cotidiana de las participantes y que pudiese emerger por medio de sus reacciones a partir de lo vivido del abuso sexual. Resultados: después de la interpretación, los resultados fueron agrupados en las categorías: Convivencia familiar después de la revelación del abuso sexual; la vida cotidiana de niñas en vivencia de abuso sexual; y repercusiones del abuso sexual en la vida adulta de mujeres abusadas sexualmente en la infancia. Las repercusiones del abuso sexual fueron descritas como dificultades en el convivio familiar, embarazo, conducta hipersexualizada, prostitución, contradicción entre género y sexo, dificultades para tener orgasmo, uso de drogas, baja autoestima, depresión, comportamiento auto-descriptivo, ideas suicidas y homicidas. Conclusión: la exposición al abuso sexual en el contexto familiar perjudicó la salud física y emocional de niñas y adolescentes, así como la convivencia familiar, aportando para la necesidad de adopción de sensibilidad y solidaridad en el cuidado a mujeres con quejas que puedan estar asociadas a vivencias de abuso sexual.

ABSTRACT Objective: to understand the repercussions of sexual abuse in the adult life of women who were sexually abused in childhood. Method: qualitative research, developed through unstructured interviews with nine women in a Reference Center for Women, in the semi-arid region of the state of Pernambuco, Brazil. The interpretation of the results was based on the approach of Comprehensive Sociology and Daily Life in order to identify the underground centrality submerged in the daily life of the participants and which could emerge through their reactions from the experience of sexual abuse. Results: after interpretation, the results were grouped into the categories: Family life after the revelation of sexual abuse; The daily life of girls experiencing sexual abuse; and Repercussions of sexual abuse in the adult life of sexually abused women in childhood. The repercussions of sexual abuse were described as difficulties in family life, pregnancy, hypersexual behavior, prostitution, gender and sex contradiction, difficulties to have orgasm, drug use, low self-esteem, depression, self-destructive behavior, suicidal ideation and homicide. Conclusion: exposure to sexual abuse in the family context has impaired the physical and emotional health of girls and adolescents, as well as family coexistence, pointing to the need to adopt sensitivity and solidarity in the care of women with complaints that may be associated with experiences of sexual abuse.

Factores bioquímicos y nutricionales asociados a la viscosidad sanguínea en adultos de la sierra urbana (Imbabura), Ecuador / Biochemical and nutritional factors associated with blood viscosity in adults living in a mountain chain, (Imbabura), Ecuador

La asociación de la viscosidad sanguínea (VS) con la hipertensión y la diabetes mellitus 2, indica que debe ser considerada sobre todo en poblaciones que viven en la sierra donde el hematocrito y la hemoglobina son más elevados. Se evaluó la asociación de la VS con factores de riesgo cardiovascular y de síndrome metabólico (SM) en una población de 237 adultos (20-60 años) de la sierra urbana ecuatoriana. Se calculó el índice de masa corporal (IMC), porcentaje de grasa y agua corporal, se midieron la circunferencia de la cintura (CC) y la presión arterial. Se realizaron determinaciones de parámetros bioquímicos, hematológicos y de VS (mediante fórmula validada). El 14,76% de la población presentó hiperviscosidad (HV). El 42,8% de las personas con HV presentó SM contra 33% en el grupo con normoviscosidad (NV). El ácido úrico (AU) se observó más elevado en los grupos HV y NV con SM. Se encontró una correlación lineal simple de las concentraciones de glucosa (GLU) y de triglicéridos (TRI) con la VS en el grupo HV sin SM. El análisis de regresión lineal múltiple indicó que la GLU, el AU y la presión sistólica son variables que influyen independientemente con el incremento de la VS en el grupo HV sin SM. Las variables GLU y % de agua influyen en el incremento de la VS en el grupo NV sin SM. El incremento de las concentraciones de glucosa y de ácido úrico son los dos factores que mayormente influyen en la VS en esta población.

The association of blood viscosity (BV) with hypertension and diabetes mellitus type 2 indicates that it should be considered, especially in populations living in mountain chains where the hemoglobin and hematocrit values are higher. In fact, this work analyzed the association of BV with nutritional and biochemical risk factors in the development of cardio metabolic diseases in healthy adults (20-60 years old; 46.39% female and 53.61% male) living in the urban Ecuadorian sierra. The body mass index (BMI) was calculated. The body fat percentage, body water percentage, waist circumference (WC) and blood pressure were measured. Determinations of biochemical and hematological parameters were performed using established methodologies and a validated formula was used to determine the BV. Hyperviscosity (HV) was present in 14.76% of the population: 10.46% men, 4.3% women. In this group, 42.8% showed metabolic syndrome (MS) against 33% in the normoviscosity group (NV). Uric acid (UA) was observed statistically higher in HV and NV with MS groups. A simple positive linear correlation was found between glucose (GLU) and triglycerides (TRI) concentrations and BV in HV without MS group. The multiple linear regression analysis indicated that GLU, UA and systolic pressure influence to increase BV in the HV without MS group. Glucose concentration and percentage of water, independently influence the BV in the NV without MS group. The increase in GLU and UA concentrations are the most influencing factors on BV in this population.

Immunogenicity assessment of HPV16/18 vaccine using the glutathione S-transferase L1 multiplex serology assay.

The glutathione S-transferase (GST)-L1 multiplex serology assay has favorable properties for use in clinical trials and epidemiologic studies, including low cost, high throughput capacity, and low serum volume requirement. Therefore, we evaluated the GST-L1 assay as a measure of HPV16/18 vaccine immunogenicity. Our study population included 65 women selected from the Costa Rica Vaccine Trial who received the bivalent HPV16/18 virus-like particle (VLP) vaccine at the recommended 0/1/6-month schedule. We tested replicate serum samples from months 0/1/12 (i.e., after 0/1/3 doses) by GST-L1 and 3 other commonly used serology assays, VLP-ELISA, SEAP-NA, and cLIA. We calculated the percentage of women seropositive by GST-L1 by time point and HPV type (14 HPV types), and compared GST-L1 to other assays using Spearman rank correlation coefficients. After 1 vaccine dose, seropositivity by GST-L1 was 40% each for HPV16 and HPV18, increasing to 100% and 98%, respectively, after 3 doses. Seropositivity after 3 doses ranged from 32% to 69% for HPV types 31/33/45, for which partial vaccine efficacy is reported, though increases also occurred for types with no evidence for cross-protection (e.g., HPV77). GST-L1 correlated best after 3 doses with VLP-ELISA (HPV16 and HPV18 each ρ = 0.72) and SEAP-NA (HPV16 ρ = 0.65, HPV18 ρ = 0.71) (all P < 0.001); correlation was lower with cLIA. The GST-L1 is suitable for evaluating HPV16/18 vaccine immunogenicity after 3 vaccine doses, although in contrast to other assays it may classify some samples as HPV16/18 seronegative. The assay's utility is limited for lower antibody levels such as after receipt of 1 dose.

Calcium carbonate precipitation by heterotrophic bacteria isolated from biofilms formed on deteriorated ignimbrite stones: influence of calcium on EPS production and biofilm formation by these isolates.

Heterotrophic CaCO3-precipitating bacteria were isolated from biofilms on deteriorated ignimbrites, siliceous acidic rocks, from Morelia Cathedral (Mexico) and identified as Enterobacter cancerogenus (22e), Bacillus sp. (32a) and Bacillus subtilis (52g). In solid medium, 22e and 32a precipitated calcite and vaterite while 52g produced calcite. Urease activity was detected in these isolates and CaCO3 precipitation increased in the presence of urea in the liquid medium. In the presence of calcium, EPS production decreased in 22e and 32a and increased in 52g. Under laboratory conditions, ignimbrite colonization by these isolates only occurred in the presence of calcium and no CaCO3 was precipitated. Calcium may therefore be important for biofilm formation on stones. The importance of the type of stone, here a siliceous stone, on biological colonization is emphasized. This calcium effect has not been reported on calcareous materials. The importance of the effect of calcium on EPS production and biofilm formation is discussed in relation to other applications of CaCO3 precipitation by bacteria.

Indicadores de saúde no cuidado ao paciente crítico neurológico / Health indicators in the care for neurocritical patients / indidores de salud en la atención al paciente crítico neurológico

O propósito deste estudo foi desenvolver indicadores para avaliação do cuidado a pacientes críticos neurológicos. Estudo metodológico de elaboração e validação de indicadores de avaliação em saúde, desenvolvido em Unidade de Terapia Intensiva de hospital de porte extra. Foi utilizada a metodologia de validação opinativa, onde sete experts (cinco enfermeiros e dois médicos) participaram da análise dos conteúdos através de manual operacional no período de maio a agosto de 2012. Os valores obtidos a partir do julgamento efetuado pelos experts foram todos acima de 80%, sendo o referencial de literatura acima de 75%, portanto os resultados indicaram que todos os indicadores foram considerados válidos em relação ao percentual de concordância. Portanto é possível o estabelecimento de indicadores confiáveis para avaliação do cuidado proposto e estabelecer a qualidade do serviço prestado. A partir de pressupostos teóricos e sua aplicação subsidiará o controle da qualidade assistencial à medida que colabora com a elaboração de plano de intervenção.

This study aimed to develop indicators to evaluate the care for neurocritical patients. Methodological study for development and validation of indicators of health evaluation, conducted on the Intensive Care Unit of a large hospital. The opinionated validation method was used, where seven experts (five nurses and two doctors) participated on the content analysis through operational manual from May to August 2012. The values obtained from the expert judgment were all above 80%, in which the literature reference was above 75%. The results revealed that all the indicators were considered valid as regards the agreement percentage. Therefore, it is possible to establish reliable indicators to evaluate the proposed care and establish the quality of the service. From the theoretical prerequisites, its use will support the control of assistance quality whilst collaborates with designing intervention plans.

El propósito del estudio fue desarrollar indicadores para evaluar la atención a pacientes críticos neurológicos. Estudio metodológico de desarrollo y validación de indicadores de evaluación en salud, llevado a cabo en Unidad de Cuidados Intensivos del hospital de franqueo adicional. Fue utilizada metodología de validación de opinión, donde participaron siete expertos (cinco enfermeros y dos médicos) en el análisis de contenidos de manual operacional de mayo a agosto de 2012. Los valores obtenidos a partir de la sentencia de los expertos estaban todos por encima de 80%, la literatura referencia por encima de 75%, por lo tanto, los resultados indicaron que los indicadores válidos en relación con el porcentaje deconcordancia. Así, es posible establecer indicadores seguros para evaluar la atención propuesta y establecer la calidad asistencial. Los presupuestos teóricos y su aplicación subvencionaran el control de la calidad asistencial, ya que colaboracon el desarrollo del plan de intervención.

A mouse air pouch model for evaluating the immune response to Taenia crassiceps infection.

The experimental system of Taenia crassiceps cysticerci infection in BALB/c mice is considered to be the most representative model of cysticercosis. In our work, mice were sacrificed 7 and 30days after infection, and pouch fluid was collected to determine the number of accumulated cells and the concentrations of IFNγ, IL-2, IL-4, IL-6, IL-10 and nitric oxide. The injection of 50 nonbudding cysticerci into normal mouse dorsal air pouches induced a high level of IFNγ and nitric oxide production relative to the parasite load. The air pouch provides a convenient cavity that allows studying the cellular immunological aspects of the T. crassiceps parasite. The nonbudding cysticerci recovered from the air pouches contained cells that can reconstitute complete cysts in the peritoneal cavity of mice. In conclusion, these results demonstrate that the air pouch model is an alternative tool for the evaluation of the immune characteristics of T. crassiceps infection.

Evidence of hepatitis A virus circulation in central Argentina: seroprevalence and environmental surveillance.

BACKGROUND: Hepatitis A virus (HAV) has shown intermediate endemicity in Argentina, but notification of clinical cases has decreased since the introduction of the vaccine in 2005. OBJECTIVES: In order to get insight into the local circulation of this virus after four years of the official introduction of the vaccine, the aims of this study were to provide information on HAV immune status of the adult population of Córdoba city and to conduct environmental surveillance of HAV in sewage and river samples in the same region. STUDY DESIGN: The prevalence of anti-HAV was determined by EIA in 416 samples of people (without prior vaccination) from Córdoba city (2009-2010). Spline regression models were estimated under generalized additive models. Environmental surveillance was conducted in river and sewage samples collected in the same period. Viral detection was performed by RT-Nested PCR of the 5'UTR. RESULTS: In Córdoba, the global prevalence of anti-HAV was 73.5%. It increased with age (p<0.0001) and it was associated with the low-income population (OR: 1.14; 95% CI 1.05-1.25). This prevalence decreased in younger age groups, especially in the high-income population. Environmental monitoring revealed the presence of HAV (IA) in 20.8% and 16.1% of wastewater and river samples, respectively. CONCLUSIONS: As a consequence of a decrease in HAV circulation due to improvements in immunization, socio-economic and hygienic conditions, young adults are becoming increasingly susceptible to HAV infections. Environmental monitoring demonstrated that HAV circulates in the local population; therefore, health care systems should consider the implementation of preventive measures for susceptible adults in order to reduce the risk of HAV infection.

Influence of occlusal contact area on cusp defection and stress distribution.

OBJECTIVE: The purpose of this study was to evaluate the effect of occlusal contact area for loading on the cuspal defection and stress distribution in a first premolar restored with a high elastic modulus restorative material. MATERIALS AND METHODS: The Rhinoceros 4.0 software was used for modeling the three-dimensional geometries of dental and periodontal structures and the inlay restoration. Thus, two different models, intact and restored teeth with three occlusal contact areas, 0.1, 0.5 and 0.75 mm(2), on enamel at the occlusal surface of buccal and lingual cusps. Finite element analysis (FEA) was performed with the program ANSYS (Workbench 13.0), which generated a mesh with tetrahedral elements with greater refinement in the regions of interest, and was constrained at the bases of cortical and trabecular bone in all axis and loaded with 100 N normal to each contact area. RESULTS: To analysis of maximum principal stress, the smaller occlusal contact area showed greater compressive stress in region of load application for both the intact and inlay restored tooth. However, tensile stresses at the occlusal isthmus were similar for all three tested occlusal contact areas (60 MPa). To displacement of the cusps was higher for teeth with inlay (0.46-0.48 mm). For intact teeth, the smaller contact area showed greater displacement (0.10 mm). For teeth with inlays, the displacement of the cusps were similar in all types of occlusal area. CONCLUSION: Cuspal displacement was higher in the restored tooth when compared to the intact tooth, but there were no significant variations even with changes in the occlusal contact area. RELEVANCE CLINICAL: Occlusal contacts have a great influence on the positioning of teeth being able to maintain the position and stability of the mandible. Axial loads would be able to generate more uniform stress at the root presenting a greater concentration of load application in the point and the occlusal surface. Thus, is necessary to analyze the relationship between these occlusal contacts as dental wear and subsequent occlusal interferences.

Pharmacological alterations that could underlie radiation-induced changes in associative memory and anxiety.

It is widely known that ionizing radiation is a physical agent broadly used to kill tumor cells during human cancer therapy. Unfortunately, adjacent normal tissues can concurrently undergo undesirable cell injury. Previous data of our laboratory demonstrated that exposure of developing rats to ionizing radiations induced a variety of behavioral differences respect to controls, including changes in associative memory and in anxiety state. However, there is a lack of data concerning modifications in different related pharmacological intermediaries. Therefore, the aim of the present study was to investigate whether the behavioral differences observed in young animals irradiated at birth might be underlain by early changes in PKCß1 levels which, in turn, could lead to changes in hippocampal GABAergic neurotransmission. Male Wistar rats were irradiated with 5Gy of X rays between 24 and 48 h after birth. Different pharmacological markers related to the affected behavioral tasks were assessed in control and irradiated hippocampus at 15 and 30 days, namely GABAA receptor, GAD65-67, ROS and PKCß1. Results showed that all measured parameters were increased in the hippocampus of 30-days-old irradiated animals. In contrast, in the hippocampus of 15-days-old irradiated animals only the levels of PKCß1 were decreased. These data suggest that PKCß1 might constitute a primary target for neonatal radiation damage on the hippocampus. Therefore, it could be hypothesized that an initial decrease in the levels of this protein can trigger a subsequent compensatory increase that, in turn, could be responsible for the plethora of biochemical changes that might underlie the previously observed behavioral alterations.

Antidepressant-like effect of α-tocopherol in a mouse model of depressive-like behavior induced by TNF-α.

Taking into account that pro-inflammatory cytokines and oxidative and nitrosative stress are implicated in the pathogenesis of depression and that α-tocopherol has antidepressant, anti-inflammatory and antioxidant properties, this study investigated the ability of α-tocopherol to abolish the depressive-like behavior induced by i.c.v. administration of TNF-α in the mouse TST. Additionally, we investigated the occurrence of changes in the levels of Bcl2 and Bax and phosphorylation of GSK-3ß (Ser9) in the hippocampus of mice. The administration of TNF-α (0.001fg/site, i.c.v.) increased the immobility time in the TST, which was prevented by the administration of α-tocopherol at the doses of 10, 30 and 100mg/kg (p.o.). Subeffective doses of α-tocopherol (10mg/kg, p.o.) and/or the antidepressants fluoxetine (5mg/kg, p.o.), imipramine (0.1mg/kg, p.o.) and bupropion (1mg/kg, p.o.), the NMDA receptor antagonist MK-801 (0.001mg/kg, p.o.) or the neuronal nitric oxide synthase inhibitor 7-nitroindazole (25mg/kg, i.p.) prevented the depressive-like effect induced by TNF-α. None of the treatments altered the locomotor activity of mice. Treatment with TNF-α and/or α-tocopherol did not alter the levels of Bax and Bcl2 or the phosphorylation of GSK-3ß in the hippocampus of mice. Together, our results show a synergistic antidepressant-like effect of α-tocopherol with antidepressants against the depressive-like behavior induced by an inflammatory insult, suggesting that this vitamin may be useful to optimize conventional pharmacotherapy of depression, including depressive states associated with inflammatory conditions.

Vih-Sida: complicaciones neurológicas de los pacientes asistidos en un hospital general de adultos / HIV-AIDS: Neurological complications of patients treated in a general hospital for adults

Resumen: Arch Med Interna 2013 - 35(1):05-08 Las complicaciones neurológicas son frecuentes en los pacientes afectados por el virus de inmunodeficiencia humana (VIH). Se realizó un estudio descriptivo, retrospectivo, para conocer la frecuencia de la patología neurológica en los pacientes VIH/SIDA (síndrome de inmunodeficiencia adquirida) de nuestro centro, su perfil clínico y evolutivo, comparar el grado de inmunodepresión, el tiempo de estadía hospitalaria, la gravedad de la afección (estimada por requerimiento de ingreso a CTI) y la mortalidad entre los pacientes que se presentaban con enfermedades oportunistas (EO) y enfermedades no oportunistas (ENO). Se encontró una alta prevalencia de enfermedad neurológica (21%), en población joven, en su mayoría de sexo masculino, con inmunodepresión muy severa, predominando la etapa SIDA; las EO son las más frecuentes y registran mayor morbimortalidad que las ENO. Nuestra población no registra la transición epidemiológica del mundo desarrollado vinculado al uso de terapia antirretroviral (TARV), hecho que atribuímos a sus condiciones socio-económico-culturales.

Abstract: Arch Med Interna 2013 - 35(1):05-08 Neurological complications are common in patients with human immunodeficiency virus (HIV). We conducted a descriptive study, to determine the frequency of neurological disease in HIV/AIDS (acquired immunodeficiency syndrome) in our center, their clinical and developmental profile, and compare the degree of immunosuppression, the length of hospital stay, severity of the condition (estimated income requirement to intensive care unit [ICU]) and mortality between the patients presenting with opportunistic disease (OD) and non-opportunistic disease (NOD). High prevalence of neurological disease (21%) was found, mostly in young people, male, with severe immunosuppression, dominating the AIDS stage. ODs are more frequent and have higher recorded morbidity than NOD. Our population does not record the epidemiological transition seen in the developed world linked to the use of antiretroviral therapy (ART), a fact we attribute to the socio-economic-cultural situation.

In vitro evaluation of the odontogenic potential of mouse undifferentiated pulp cells

The aim of this study was to evaluate the odontogenic potential of undifferentiated pulp cells (OD-21 cell line) through chemical stimuli in vitro. Cells were divided into uninduced cells (OD-21), induced cells (OD-21 cultured in supplemented medium/OD-21+OM) and odontoblast-like cells (MDPC-23 cell line). After 3, 7, 10 and 14 days of culture, it was evaluated: proliferation and cell viability, alkaline phosphatase activity, total protein content, mineralization, immunolocalization of dentin matrix acidic phosphoprotein 1 (DMP1), alkaline phosphatase (ALP) and osteopontin (OPN) and quantification of genes ALP, OSTERIX (Osx), DMP1 and runt-related transcription factor 2 (RUNX2) through real-time polymerase chain reaction (PCR). Data were analyzed by Kruskal-Wallis and Mann-Whitney U tests (p<0.05). There was a decrease in cell proliferation in OD-21 + OM, whereas cell viability was similar in all groups, except at 7 days. The amount of total protein was higher in group OD-21 + OM in all periods; the same occurred with ALP activity after 10 days when compared with OD-21, with no significant differences from the MDPC-23 group. Mineralization was higher in OD-21+OM when compared with the negative control. Immunolocalization demonstrated that DMP1 and ALP were highly expressed in MDPC-23 cells and OD-21 + OM cells, whereas OPN was high in all groups. Real-time PCR revealed that DMP1 and ALP expression was higher in MDPC-23 cell cultures, whereas RUNX2 was lower for these cells and higher for OD-21 negative control. Osx expression was lower for OD-21 + OM. These results suggest that OD-21 undifferentiated pulp cells have odontogenic potential and could be used in dental tissue engineering.

O objetivo foi avaliar o potencial odontogênico de células indiferenciadas da polpa (OD-21) por meio de indução química in vitro. As células foram divididas em grupos: controle (OD-21), induzido (OD-21 em meio suplementado/OD-21 + OM), e células odontoblastóides (MDPC-23). Após 3, 7, 10 e 14 dias, avaliou-se proliferação e viabilidade celular, proteína total e fosfatase alcalina (ALP), mineralização, imunolocalização da proteína da matriz dentinária 1 (DMP1), ALP e osteopontina (OPN), assim como a expressão dos genes ALP, OSTERIX (Osx), DMP1 e fator de transcrição RUNX2 por PCR em tempo real. Os dados foram avaliados pelo teste de Kruskal-Wallis seguido pelo teste de Mann-Whitney U (p<0.05). Houve diminuição na proliferação celular em OD-21 + OM, com viabilidade celular similar em todos os grupos, exceto aos sete dias. O conteúdo de proteína total foi maior no grupo OD-21 + OM em todos os períodos; o mesmo ocorreu com a atividade de ALP quando comparada com o grupo OD-21, além de apresentar resultados similares ao grupo MDPC-23. A mineralização foi maior em OD-21 + OM quando comparada com o controle negativo. A imunolocalização demonstrou expressão de DMP1 e ALP em MDPC-23 e OD-21 + OM, enquanto que todos os grupos foram positivos para OPN. A expressão gênica de DMP1 e ALP foi maior nas culturas de MDPC-23, enquanto que a de RUNX2 foi menor para estas células e maior no controle negativo. A expressão de OSTERIX foi menor em OD-21 + OM quando comparada aos outros grupos. Sugere-se que as células indiferenciadas da polpa da linhagem OD-21 apresentam potencial odontogênico e poderiam ser usadas para a engenharia tecidual.

Antimicrobial activity of Syzygium cumini (Myrtaceae) leaves extract

The antimicrobial activity of Syzygium cumini leaves extract, known as "jambolão", was evaluated. The crude hydroalcoholic extract was active against Candida krusei (inhibition zone of 14.7 ± 0.3 mm and MIC = 70 µg/mL), and against multi-resistant strains of Pseudomonas aeruginosa, Klebsiella pneumoniae and Staphylococcus aureus.

A atividade antimicrobiana do extrato das folhas de Syzygium cumini (Myrtaceae), conhecido como "jambolão", foi avaliada. O extrato hidroalcoólico mostrou atividade contra Candida krusei (halo de inibição de 14.7 ± 0.3 mm e CIM = 70 µg/mL) e cepas multiresistentes de Pseudomonas aeruginosa, Klebsiella pneumoniae e Staphylococcus aureus.