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There is growing recognition across broad sectors of the scientific community that use of genomic biomarkers has the potential to reduce the need for conventional rodent carcinogenicity studies of industrial chemicals, agrochemicals, and pharmaceuticals through a weight-of-evidence approach. These biomarkers fall into 2 major categories: (1) sets of gene transcripts that can identify distinct tumorigenic mechanisms of action; and (2) cancer driver gene mutations indicative of rapidly expanding growth-advantaged clonal cell populations. This call-to-action article describes a collaborative approach launched to develop and qualify biomarker gene expression panels that measure widely accepted molecular pathways linked to tumorigenesis and their activation levels to predict tumorigenic doses of chemicals from short-term exposures. Growing evidence suggests that application of such biomarker panels in short-term exposure rodent studies can identify both tumorigenic hazard and tumorigenic activation levels for chemical-induced carcinogenicity. In the future, this approach will be expanded to include methodologies examining mutations in key cancer driver gene mutation hotspots as biomarkers of both genotoxic and nongenotoxic chemical tumor risk. Analytical, technical, and biological validation studies of these complementary genomic tools are being undertaken by multisector and multidisciplinary collaborative teams within the Health and Environmental Sciences Institute. Success from these efforts will facilitate the transition from current heavy reliance on conventional 2-year rodent carcinogenicity studies to more rapid animal- and resource-sparing approaches for mechanism-based carcinogenicity evaluation supporting internal and regulatory decision-making.

La gulupa (Passiflora edulis Sims) es una fruta exótica de sabor dulce y ligeramente ácido, aroma agradable y de gran aceptación principalmente en mercados europeos. En Colombia, una parte de las cosechas producidas está conformada por frutos que no cumplen con los estándares de calidad para la exportación, por lo que se destinan para el consumo nacional, y son comercializados a precios muy bajos; por eso, es necesario proponer alternativas para su aprovechamiento y potencial mercado. El objetivo del presente trabajo fue evaluar la influencia del grado de madurez de gulupa sobre la aceptación sensorial en productos alimenticios. Se elaboraron yogur y barras de cereal endulzados con salsa de gulupa proveniente de frutos con diferentes estados de madurez (T1: 30 %, T2: 50 %, T3: 70 % y T4: 100 %). Estos productos fueron evaluados sensorialmente por un panel de 101 consumidores, de edades entre los 18 y 54 años, por medio de la escala hedónica facial de 5 puntos. Los resultados obtenidos mostraron que el 56.44% de los consumidores mencionaron que les encantó el tratamiento T4 (yogur), y el 51.49% de los consumidores que les gustó el tratamiento T3 (barras de cereal). Lo anterior permite concluir que para el yogur, lo más recomendable es endulzar con frutas maduras, o incluso sobremaduras, mientras que para las barras de cereal lo ideal es usar gulupas con menor estado de maduración (70%).

The metabolic syndrome (MS) is a constellation of related factors that increases the risk of developing cardiovascular diseases. Vaccinium meridionale Swartz contains polyphenols that could modulate some components of MS. Epidemiological and intervention studies have shown differences between men and women in MS components and antioxidant capacity. The objective of this study is to compare between men and women with MS the effects of agraz consumption on insulin resistance, antioxidant capacity, and markers of oxidation and inflammation. Men and women diagnosed with MS according to the Adult Treatment Panel III criteria were recruited in a double-blind, crossover study of 12 weeks. Participants were assigned to consume agraz nectar or placebo over 4 weeks. After 4 weeks of washout, they were switched to the alternative treatment. At the end of each period, the components of the MS, insulin resistance, antioxidant capacity, and some oxidative (oxidized low-density lipoprotein [oxLDL]; thiobarbituric acid reactive substances) and inflammatory (high-sensitive C-reactive protein [hs-CRP]) markers were evaluated. After consuming agraz, there was a tendency to increase the levels of antioxidants and to reduce the levels of hs-CRP in both genders. In addition, women who increased their serum phenols after consuming agraz had a significant reduction in insulin resistance, which was different from the results in men. Regarding men, those who increased their serum antioxidant capacity after consuming agraz had a better effect on the reduction of oxLDL levels that was significant compared to women. There are important differences between genders in the effects of agraz consumption in adults with MS.

Under ICH M7, impurities are assessed using the bacterial reverse mutation assay (i.e., Ames test) when predicted positive using in silico methodologies followed by expert review. N-Nitrosamines (NAs) have been of recent concern as impurities in pharmaceuticals, mainly because of their potential to be highly potent mutagenic carcinogens in rodent bioassays. The purpose of this analysis was to determine the sensitivity of the Ames assay to predict the carcinogenic outcome with curated proprietary Vitic (n = 131) and Leadscope (n = 70) databases. NAs were selected if they had corresponding rodent carcinogenicity assays. Overall, the sensitivity/specificity of the Ames assay was 93-97% and 55-86%, respectively. The sensitivity of the Ames assay was not significantly impacted by plate incorporation (84-89%) versus preincubation (82-89%). Sensitivity was not significantly different between use of rat and hamster liver induced S9 (80-93% versus 77-96%). The sensitivity of the Ames is high when using DMSO as a solvent (87-88%). Based on the analysis of these databases, the Ames assay conducted under OECD 471 guidelines is highly sensitive for detecting the carcinogenic hazards of NAs.

Objective:To know the anti-infective effectiveness of ascorbic acid, after a linear incision in the mucosa of the mandibular antero-inferior segment of the oral cavity of the guinea pigs, housed in the vivarium of the Biology Center of the Central University of Ecuador. Materials and methods:This study is experimental in-vivo and longitudinal, where an incision was made in the mucosa of the mandibular anteroinferior segment of the oral cavity in guinea pigs, three groups of 10 guinea pigs were formed, Group C, received mega doses ( 1.42 ml / 214.2 mg) which contained high amounts of ascorbic acid, Group B received normal doses (0.095 ml / 14.25 mg) and group Aor control group did not receive ascorbic acid, this dose was applied one day before, during, and one day after the incision. The clinical evaluation was performed at 24 and 72 hours post incision, a sample was taken by swab from the wounded area at 72 hours for the planting of bacterial cultivation. Results: The clinical examination shows that at 24 hours in group C, 0% bleeding, edema, and erythema, and 100% epithelial junction, while in group B, 30% bleeding, 50% edema, 50% erythema and 100% epithelial junction and in group A50% bleeding, 40% edema, 40% erythema and 20% epithelial junction; at 72 hours the results in group C and B showed better healing percentages in relation to group A, in this group some guinea pigs still showed bleeding edema and erythema in the wounded area. Conclusion: It is determined that the guinea pigs who received mega doses of ascorbic acid presented a better result in the process of healing after clinical examination at 24 and 72 hours and the swab exam shows a lower amount of bacterial colonization, thus demonstrating the effectiveness of ascorbic acid in mega doses of infectious processes.

Resumen La criptococosis meníngea presenta alta mortalidad mundial, especialmente en población VIH/sida. La OMS recomienda detectar el antígeno capsular de Crypto coccus como estrategia para un diagnóstico temprano y poder minimizar complicaciones. Objetivo: realizar antigenemia temprana de Cryptococcus mediante in munocromatografía/ensayo de flujo lateral en pacientes asintomáticos VIH+. Material y método: estudio descriptivo observacional; entre julio-2016 y mayo-2019 se procesaron mediante ensayo de flujo lateral, muestras de suero de 169 pacientes asintomáticos VIH+, con CD4 ≤120 cel/μL en Barranquilla, Colombia. Ante resultado positivo, se indicó profilaxis con fluconazol; se hizo seguimiento a todos los casos. Resultados: la antigenemia fue positiva en cinco pacientes (2,96%); uno falleció, cuatro recibieron profilaxis y la prueba se negativizó en dos. Los pacientes con resultado negativo inicial no desarrollaron durante el estudio sinto matología compatible con esta micosis. Discusión: el ensayo de flujo lateral de Cryptococcus está recomendado para el diagnóstico temprano de la criptococosis en población VIH/sida. Conclusión: detectar tempranamente el antígeno circulante de Cryptococcus mediante ensayo de flujo lateral en pacientes asintomáticos VIH+, permitió instaurar profilaxis oportuna, hacer seguimiento y control para reducir la mortalidad asociada con la criptococosis meníngea. Abstract Meningeal cryptococcosis presents high levels of global mortality, especially in the HIV/AIDS population. The WHO recommends detecting the capsular antigen as an important strategy for early diagnosis and be able to minimize complications. Objective: Perform early cryptococcal antigenemia by immunochromatographic/ lateral flow assay in asymptomatic HIV+ patients. Material and method: descriptive observational study; between July-2016 and May-2019, serum samples from 169 asymptomatic HIV+ patients with CD4 ≤120 cells/μL were processed by lateral flow assay in Barranquilla, Colombia. Given a positive result, prophylaxis with fluconazole was indicated; all cases were followed up. Results: antigenemia was positive in five (2.96%) patients; one died; four received prophylaxis, and the test turned negative in two. The patients with an initial negative result, did not developed symptoms compatible with this mycosis during the study period. Discussion: lateral flow assay for Cryptococcus is recommended for the early diagnosis of cryptococcosis in the HIV/AIDS population. Conclusion: early detection of circulating Cryptococcus antigen by lateral flow assay in HIV+ patients allowed the establishment of timely prophylaxis, follow-up, and control to reduce mortality associated with meningeal cryptococcosis.

COVID-19 morbidity is highest in Black and Latino older adults. These racial and ethnic groups initially had lower vaccination uptake than others, and rates in Black adults continue to lag.To evaluate the effect of outreach via electronic secure messages and mailings from primary care physicians (PCPs) on COVID-19 vaccination uptake among Black and Latino older adults and to compare the effects of culturally tailored and standard PCP messages.This randomized clinical trial was conducted from March 29 to May 20, 2021, with follow-up surveys through July 31, 2021. Latino and Black individuals aged 65 years and older from 4 Kaiser Permanente Northern California (KPNC) service areas were included. Data were analyzed from May 27, 2021, to September 28, 2021.Individuals who had not received COVID-19 vaccination after previous outreach were randomized to electronic secure message and/or mail outreach from their PCP, similar outreach with additional culturally tailored content, or usual care. Outreach groups were sent a secure message or letter in their PCP's name, followed by a postcard to those still unvaccinated after 4 weeks.The primary outcome was time to receipt of COVID-19 vaccination during the 8 weeks after initial study outreach. KPNC data were supplemented with state data from external sources. Intervention effects were evaluated via proportional hazards regression.Of 8287 included individuals (mean [SD] age, 72.6 [7.0] years; 4665 [56.3%] women), 2434 (29.4%) were Black, 3782 (45.6%) were Latino and preferred English-language communications, and 2071 (25.0%) were Latino and preferred Spanish-language communications; 2847 participants (34.4%) had a neighborhood deprivation index at the 75th percentile or higher. A total of 2767 participants were randomized to culturally tailored PCP outreach, 2747 participants were randomized to standard PCP outreach, and 2773 participants were randomized to usual care. Culturally tailored PCP outreach led to higher COVID-19 vaccination rates during follow-up compared with usual care (664 participants [24.0%] vs 603 participants [21.7%]; adjusted hazard ratio (aHR), 1.22; 95% CI, 1.09-1.37), as did standard PCP outreach (635 participants [23.1%]; aHR, 1.17; 95% CI, 1.04-1.31). Individuals who were Black (aHR, 1.19; 95% CI, 1.06-1.33), had high neighborhood deprivation (aHR, 1.17; 95% CI, 1.03-1.33), and had medium to high comorbidity scores (aHR, 1.19; 95% CI, 1.09-1.31) were more likely to be vaccinated during follow-up.This randomized clinical trial found that PCP outreach using electronic and mailed messages increased COVID-19 vaccination rates among Black and Latino older adults.ClinicalTrials.gov Identifier: NCT05096026.

Purple passionfruit (Passiflora edulis Sims) is an exotic, sweet and slighty acid flavored fruit with a pleasant aroma. It is also widely accepted in European markets. In Colombia, a part of the yield crop is comprised of fruit that does not meet quality standards for export and is intended for national consumption, and marketed at very low prices. Therefore, it is necessary to propose alternatives for its use and potential market. The aim of this study was to assess the influence of the degree of ripeness for purple passionfruit on the sensory acceptance in food products. Yogurt and cereal bars sweetened with purple passionfruit sauce from fruit with different stages of ripeness (T1: 30 %, T2: 50 %, T3: 70 % and T4: 100 %) were prepared. By a panel of 101 consumers aged 18-54 years old, using a 5-point facial hedonic scale, said products were sensorially assessed. The results obtained showed that 56.44% of consumers preferred T4 treatment (yogurt) and 51.49% of consumers preferred T3 treatment (cereal bars). The above mentioned leads to conclude that, for yogurt preparations, it is best to sweeten with ripe or even overripe fruit, while for cereal bars it is ideal to use purple passionfruit with a lower stage of ripeness (70%)., La gulupa (Passiflora edulis Sims) es una fruta exótica de sabor dulce y ligeramente ácido, aroma agradable y de gran aceptación principalmente en mercados europeos. En Colombia, una parte de las cosechas producidas está conformada por frutos que no cumplen con los estándares de calidad para la exportación, por lo que se destinan para el consumo nacional, y son comercializados a precios muy bajos; por eso, es necesario proponer alternativas para su aprovechamiento y potencial mercado. El objetivo del presente trabajo fue evaluar la influencia del grado de madurez de gulupa sobre la aceptación sensorial en productos alimenticios. Se elaboraron yogur y barras de cereal endulzados con salsa de gulupa proveniente de frutos con diferentes estados de madurez (T1: 30 %, T2: 50 %, T3: 70 % y T4: 100 %). Estos productos fueron evaluados sensorialmente por un panel de 101 consumidores, de edades entre los 18 y 54 años, por medio de la escala hedónica facial de 5 puntos. Los resultados obtenidos mostraron que el 56.44% de los consumidores mencionaron que les encantó el tratamiento T4 (yogur), y el 51.49% de los consumidores que les gustó el tratamiento T3 (barras de cereal). Lo anterior permite concluir que para el yogur, lo más recomendable es endulzar con frutas maduras, o incluso sobremaduras, mientras que para las barras de cereal lo ideal es usar gulupas con menor estado de maduración (70%).

The in vitro MultiFlow® DNA Damage Assay multiplexes γH2AX, p53, phospho-histone H3, and polyploidization biomarkers into a single flow cytometric analysis. The current report describes a tiered sequential data analysis strategy based on data generated from exposure of human TK6 cells to a previously described 85 chemical training set and a new pharmaceutical-centric test set (n = 40). In each case, exposure was continuous over a range of closely spaced concentrations, and cell aliquots were removed for analysis following 4 and 24 hr of treatment. The first data analysis step focused on chemicals' genotoxic potential, and for this purpose, we evaluated the performance of a machine learning (ML) ensemble, a rubric that considered fold increases in biomarkers against global evaluation factors (GEFs), and a hybrid strategy that considered ML and GEFs. This first tier further used ML output and/or GEFs to classify genotoxic activity as clastogenic and/or aneugenic. Test set results demonstrated the generalizability of the first tier, with particularly good performance from the ML ensemble: 35/40 (88%) concordance with a priori genotoxicity expectations and 21/24 (88%) agreement with expected mode of action (MoA). A second tier applied unsupervised hierarchical clustering to the biomarker response data, and these analyses were found to group certain chemicals, especially aneugens, according to their molecular targets. Finally, a third tier utilized benchmark dose analyses and MultiFlow biomarker responses to rank genotoxic potency. The relevance of these rankings is supported by the strong agreement found between benchmark dose values derived from MultiFlow biomarkers compared to those generated from parallel in vitro micronucleus analyses. Collectively, the results suggest that a tiered MultiFlow data analysis pipeline is capable of rapidly and effectively identifying genotoxic hazards while providing additional information that is useful for modern risk assessments-MoA, molecular targets, and potency. Environ. Mol. Mutagen. 60:513-533, 2019. © 2019 Wiley Periodicals, Inc.

In a previously reported CD-1 mouse 2-year carcinogenicity study with the sodium glucose cotransporter-2 inhibitor empagliflozin, an increased incidence of renal tubular adenomas and carcinomas was identified only in the male high-dose group. Follow-up investigative studies have shown that the renal tumors in male high-dose mice were preceded by a number of renal degenerative/regenerative findings. Prior cross-species in vitro metabolism studies using microsomes identified an oxidative metabolite (M466/2) predominantly formed in the male mouse kidney and which spontaneously degrades to a metabolite (M380/1) and reactive 4-OH crotonaldehyde (CTA). In order to further evaluate potential modes of action for empagliflozin-associated male mouse renal tumors, we report here a series of in vitro investigative toxicology studies conducted to evaluate the cytotoxic and genotoxic potential of empagliflozin and M466/2. To assess the cytotoxic potential of empagliflozin and M466/2, a primary mouse renal tubular epithelial (mRTE) cell model was used. In mRTE cells, M466/2-derived in vitro 4-OH CTA exposure was cytotoxic, while empagliflozin was not cytotoxic or mitogenic. Empagliflozin and M466/2 were not genotoxic, supporting an indirect mode of action for empagliflozin-associated male mouse renal tumorigenesis. In conclusion, these in vitro data show that M466/2-derived 4-OH CTA exposure is associated with cytotoxicity in renal tubule cells and may be involved in promoting compound-related in vivo renal metabolic stress and chronic low-level renal injury, in turn supporting a nongenotoxic mode of tumor pathogenesis specific to the male mouse.

espanolIntroduccion: Las sustancias relacionadas con los microorganismos involucrados en la enfermedad periodontal puedan llegar a la interfaz materno-fetal por via hematogena y estimular la contractilidad uterina. Objetivo: Determinar la asociacion entre enfermedad periodontal con nacimiento pretermino. Metodo: Estudio de casos y controles de 343 embarazadas pretermino y 686 de termino. Se calculo la edad gestacional por fecha de ultimo periodo menstrual y se confirmo con los metodos de Capurro y Ballard. La enfermedad periodontal se diagnostico por la profundidad del espacio entre la raiz dental y la encia. La asociacion fue medida con regresion logistica. Resultados: La edad de las madres en los casos fue de 23.8 ± 6.7 anos y en los controles de 23.2 ± 6.7 anos. La enfermedad periodontal estuvo presente en 66.8 % de los casos y 40.5 % de los controles. Los factores asociados con nacimiento pretermino fueron enfermedad periodontal (RM = 2.26), antecedente de nacimiento pretermino (RM = 4.96), embarazo no planeado (RM = 2.15), control prenatal deficiente (RM = 2.53), infeccion de vias urinarias (RM = 2.22), preeclampsia (RM = 4.49), ruptura prematura de membranas amnioticas (RM = 2.59) y nacer por cesarea (RM = 9.15). Conclusion: La enfermedad periodontal en el embarazo constituyo un factor de riesgo independiente para nacimiento pretermino. EnglishIntroduction: Substances related to microorganisms involved in periodontal disease can reach the maternal-fetal interface via the hematogenous route and stimulate uterine contractility. Objective: To determine the association between periodontal disease and preterm birth. Method: Case-control study in 343 preterm and 686 full-term pregnant women. Gestational age was calculated based on the date of the last menstrual period and confirmed with Capurro and Ballard methods. Periodontal disease was diagnosed according to the depth of the space between the tooth root and the gum. The association was measured with logistic regression. Results: Maternal age of the cases was 23.8 ± 6.7 years, and 23.2 ± 6.7 in the controls. Periodontal disease was present in 66.8% of cases and 40.5% of controls. The factors associated with preterm birth were periodontal disease (Odds ratio [OR] = 2.26), history of preterm birth (OR = 4.96), unplanned pregnancy (OR = 2.15) poor prenatal control (OR = 2.53), urinary tract infection (OR = 2.22), preeclampsia (OR = 4.49), premature rupture of membranes (OR = 2.59) and caesarean section delivery (OR = 9.15). Conclusion: Periodontal disease in pregnancy was an independent risk factor for preterm birth.

The bacterial reverse mutation test is a mainstay for evaluation of mutagenicity predicting the carcinogenic potential of a test substance and is recommended by regulatory agencies across the globe. The popularity of the test is due, in part, to the relatively low cost, rapid results and small amount of test material required compared to most other toxicological tests as well as the near universal acceptance of the toxicological significance of a clear positive or negative result. Most laboratories follow the Organization for Economic Cooperation and Development Test Guideline 471 (TG471) or national guidelines based on TG471. Regulatory agencies in most countries are obligated to consider results from tests which meet the recommendations laid out in TG471. Nonetheless, laboratories unfamiliar with the test sometimes have trouble generating reliable, reproducible results. TG471 is a test guideline, not a detailed test protocol. A group of experts from regulatory agencies and laboratories which use the assay has assembled here a set of recommendations which if followed, will allow an inexperienced laboratory to acquire proficiency in assay conduct. These include recommendations for how to create a cell bank for the 5 Salmonella typhimurium/Escherichia coli strains and develop a laboratory protocol to reliably culture each strain to ensure each culture has the characteristics which allow adequate sensitivity for detection of mutagens using the test as described in TG471. By testing compounds on the provided lists of positive and negative test substances, the laboratory will have surmounted many of the problems commonly encountered during routine testing of unknown chemicals and will have gained the experience necessary to prepare the detailed protocol needed for performing the test under Good Laboratory Procedures and the laboratory will have generated the historical positive and negative control databases which are needed for test reports which adhere to TG471.

A database of 91 chemicals with published data from both transgenic rodent mutation (TGR) and rodent comet assays has been compiled. The objective was to compare the sensitivity of the two assays for detecting genotoxicity. Critical aspects of study design and results were tabulated for each dataset. There were fewer datasets from rats than mice, particularly for the TGR assay, and therefore, results from both species were combined for further analysis. TGR and comet responses were compared in liver and bone marrow (the most commonly studied tissues), and in stomach and colon evaluated either separately or in combination with other GI tract segments. Overall positive, negative, or equivocal test results were assessed for each chemical across the tissues examined in the TGR and comet assays using two approaches: 1) overall calls based on weight of evidence (WoE) and expert judgement, and 2) curation of the data based on a priori acceptability criteria prior to deriving final tissue specific calls. Since the database contains a high prevalence of positive results, overall agreement between the assays was determined using statistics adjusted for prevalence (using AC1 and PABAK). These coefficients showed fair or moderate to good agreement for liver and the GI tract (predominantly stomach and colon data) using WoE, reduced agreement for stomach and colon evaluated separately using data curation, and poor or no agreement for bone marrow using both the WoE and data curation approaches. Confidence in these results is higher for liver than for the other tissues, for which there were less data. Our analysis finds that comet and TGR generally identify the same compounds (mainly potent mutagens) as genotoxic in liver, stomach and colon, but not in bone marrow. However, the current database content precluded drawing assay concordance conclusions for weak mutagens and non-DNA reactive chemicals.

This case report describes a technique in which endodontic treatment and permanent indirect restoration were completed in the same clinical appointment with the aid of a computer-aided design/computer-aided manufacturing (CAD/CAM) system. Two patients were diagnosed with irreversible pulpitis of the mandibular first molar. After access preparation, root canals were located, irrigation was performed until bleeding ceased, and the coronal tooth structure was prepared for indirect restoration. Then, utilizing an interim 3-mm build-up of the endodontic access cavity, a hemi-arch digital scan was performed with an intraoral scanner. Subsequent to digital scanning, restoration design was performed simultaneously with the endodontic procedure. The root canals were shaped using the Race system under irrigation with 2.5% sodium hypochlorite followed by root canal filling. The pulp chamber was subsequently filled with a 3-mm-thick composite resin restoration mimicking the interim build-up previously utilized to facilitate block milling in the CAD/CAM system. Clinical try-in of the permanent onlay restoration was followed by acid etching, application of a 5th generation adhesive, and cementation of the indirect restoration. Once the restoration was cemented, rubber dam isolation was removed, followed by occlusal adjustment and polishing. After 2 years of follow-up, the restorations were esthetically and functionally satisfactory, without complications.

BACKGROUND Health information technology (IT) tools are increasingly used to improve patient care. However, implementation of English-only health IT tools could potentially worsen health disparities for non-English speakers. OBJECTIVE We aim to describe the “trans-creation” process of developing linguistically and culturally appropriate health IT tools through a detailed case analysis of a waiting room health mobile app designed to help Spanish-speaking Latino people prepare for primary care visits. METHODS We adapted the English-language Visit Planner mobile app for Spanish-speaking Latino patients. We applied culturally defined themes derived from prior published research and input by both skilled linguists and potential end users. Initial changes were iteratively reviewed and edited by a team of writers, health care educators, subject matter experts, patients, and providers. RESULTS The trans-creation process resulted in the following key culturally mediated changes to the tool: replacing the “provider” actors with “patient” actors; changing the choice of “Stress at Home or Work” (represented by an icon of a house) to “Mi Familia” (translation: my family; icon is an outline of family members holding hands); replacing the English terms “anxiety” and “depression” with “Me siento desanimado”(translation: I am feeling down) to avoid mental health stigma; and using more concise text translation to ensure the wording fit the available on-screen space. CONCLUSIONS The trans-creation process of cultural and linguistic adaptation led to several design changes that would not have been implemented if we had simply translated the words from English to Spanish.

A committee was constituted within the International Workshop on Genetic Toxicology Testing (IWGT) to evaluate the current criteria for a valid Ames test and to provide recommendations for interpretation of test results. Currently, determination of a positive vs. a negative result is made by applying various data evaluation procedures for comparing dosed plates with the concurrent solvent control plates. These evaluation procedures include a requirement for a specific fold increase (2- or 3-fold, specific to the bacterial strain), formal statistical procedures, or subjective (expert judgment) evaluation. After extensive discussion, the workgroup was not able to reach consensus recommendations in favor of any of these procedures. There was a consensus that combining additional evaluation criteria to the comparison between dosed plates and the concurrent solvent control plates improves test interpretation. The workgroup recommended using these additional criteria because the induction of mutations is a continuum of responses and there is no biological relevance to a strict dividing line between a positive (mutagenic) and not-positive (nonmutagenic) response. The most useful additional criteria identified were a concentration-response relationship and consideration of a possible increase above the concurrent control in the context of the laboratory’s historical solvent control values for the particular tester strain. The workgroup also emphasized the need for additional testing to resolve weak or inconclusive responses, usually with altered experimental conditions chosen based on the initial results. Use of these multiple criteria allowed the workgroup to reach consensus on definitions of “clear positive” and “clear negative” responses which would not require a repeat test for clarification. The workgroup also reached consensus on recommendations to compare the responses of concurrent positive and negative controls to historical control distributions for assay acceptability, and the use of control charts to determine the validity of the individual test.

Este trabajo tiene por objetivo mostrar cómo se han tejido algunos entramados sociales, culturales y académicos alrededor de la conformación de una nueva sociedad, la sociedad de la información, mediada por lo informacional y generada principalmente por la aparición de la internet. Destaca el artículo los apuntes de referencia al uso de la web con fines académicos a partir del manejo de los rastreos bibliográficos. Se pretende, además, que los maestros pongan la mirada en algunos principios propios del quehacer docente como son la enseñanza, la puesta en práctica del currículo oculto y el manejo de la didáctica para el uso de la información virtual en el aula.Palabras clave: rastreo bibliográfico, ciberespacio, autopista electrónica, investigación, realidad virtual.

Las macrófitas acuáticas han sido utilizadas en procesos de biorremediación, ya que pueden absorber algunas sustancias disueltas y producir oxígeno mediante fotosíntesis. Con base en los estudios de remoción de compuestos tóxicos por parte de plantas acuáticas que permitan la depuración de aguas residuales, la presente investigación tuvo como propósito principal buscar la disminución de la contaminación por cianuro y la toxicidad de las aguas residuales del lavado de arena del proceso de cianuración de oro de la mina La Coqueta, mediante la utilización de macrófitas acuáticas flotantes (Eichhornia crassipes,Pistia stratiotesSalvinia auriculata); además, de acuerdo con los resultados de pruebas cualitativas, encontrar el destino final del cianuro en las plantas al ser absorbidos por éstas. El tratamiento de agua cianurada se realizó mediante un sistema de acuarios, los cuales contenían el agua proveniente de la mina, en ellos se depositaron las plantas para el tratamiento, se tomaron parámetros fisicoquímicos de control (pH, porcentaje de saturación de oxígeno y conductividad), además de la determinación de cianuro y toxicidad en el agua, pruebas cualitativas de compuestos nitrogenados en las plantas y bioensayos conDaphnia pulexen las plantas y en el agua tratada. Con base en los resultados de las pruebas cualitativas, se evidenció que las macrófitas pueden cambiar su metabolismo en condiciones de estrés y este cambio de metabolismo induce también a que se produzcan otro tipo de metabolitos secundarios no presentes naturalmente (e. g. glicósidos cianogénicos y alcaloides) en las plantas sometidas al tratamiento.La remoción de cianuro por parte de estas especies fue hasta del 86% al final del tratamiento, demostrando gran eficiencia para remover compuestos tóxicos del agua cianurada proveniente de la mina La Coqueta, ya que en los bioensayos realizados conDaphnia pulexindicaron que estos organismos podían resistir a las características del agua después del tratamiento con las plantas. Señalando que se puede realizar un tratamiento natural, ambientalmente seguro y económicamente viable que permita descontaminar el agua cianurada proveniente del lavado de las arenas del beneficio del oro a través del proceso de cianuración. Aquatic macrophytes have been used in bioremediation processes, as they can absorb some dissolved substances and produce oxygen through photosynthesis. Based on the studies of toxic compounds removal by aquatic plants that allow wastewater treatment, this research had as main purpose to search the reduction of cyanide contamination and toxicity of waste water from sand washing in the cyanidation process inLa Coquetagold mine by means of the use of floating aquatic macrophytes (Eichhornia crassipes, Salvinia auriculata, Pistiastratiotes). Similarly, according to the results of qualitative tests, a goal was to find the final destiny of cyanide afterplants had absorbed it. Treatment of cyanide water was conducted through a system of tanks containing water from the mine in which plants for treatment were included and physicochemical control parameters (pH, oxygen saturation percentage and conductivity) were taken. Besides determination of cyanide and water toxicity, qualitative testsof nitrogenous compounds in plants and bioassays withDaphnia pulexin plants and in the treated water were determined. Based on the results of qualitative tests ti was evidenced that macrophytes can change their metabolism under stress conditions and this change in metabolism also induces the production of other types of secondary metabolites not present naturally (e.g. cyanogenic glycosides and alkaloids) in plants subjected to treatment. The removal of cyanide by these species was up to 86% at end of treatment, showing high efficiency for removal of toxic compounds of cyanide water fromLa Coquetamine, as bioassays conducted usingDaphnia pulexindicated that these organism could resist water characteristics after treatment with plants, noting that natural treatment environmentally safe and economically feasible can be made to decontaminate cyanide water from the washing of sands resulting from cyanide gold processing.

Acute myeloid leukaemia (AML) is associated with exposure to benzene and treatment with chemotherapeutic agents. It is thought to arise from damage to specific regions of DNA, resulting in chromosome rearrangements or loss. For instance, a deletion on the long arm of chromosome 5 [e.g. del(5q31)] is common in AML patients previously treated with alkylating agents, such as melphalan, or exposed to benzene. Translocations of the MLL gene at 11q23 are frequently observed in AML arising from treatment with topoisomerase II inhibitors, such as etoposide. Our goal was to determine whether or not breakage at 5q31 and 11q23 is selectively induced by these chemical agents. To address this question, the comet assay combined with fluorescence in situ hybridization (comet-FISH) was used to detect DNA breakage in the specific chromosomal regions in an in vitro model. TK6 lymphoblastoid cells were exposed to melphalan, etoposide or the benzene metabolite, hydroquinone (HQ), at various concentrations. HQ, melphalan and etoposide induced DNA breaks at both 5q31 and 11q23 chromosome regions in a dose-dependant manner. However, HQ produced significantly more DNA damage at 5q31 than at 11q23. Etoposide produce slightly more DNA damage at 11q23 and melphalan had a somewhat greater effect at 5q31, but not significantly so. Thus, HQ and melphalan act similarly, perhaps explaining some similarities between benzene- and alkylating agent-induced AML. Comet-FISH also appears to be a useful approach for detecting and comparing damage to specific chromosome regions of significance in leukaemogenesis.

Cuando hablamos de las “rebeldías de los maestros”, lo hacemos desde el proceso interior del ser educador, nos referimos a aquellas posturas crí-ticas y reveladoras, exigiéndonos transitar por varios escenarios: nuestras autobiografías que han develado las huellas epocales y rebeldías que de una u otra forma se conectan con diferentes historias de maestros; y de acontecimientos transcurridos a partir de la década de los 60, en donde se manifiesta una realidad política, económica, educativa, y religiosa, trans-versalidas por el desarrollo y las experiencias como maestros; el exilio, el silencio, las rebeldías, el revelarse, la desesperanza y el poder, aspectos que marcan el camino recorrido y por recorrer, levantando códigos y ca-tegorías que influyen en el quehacer docente; la innovación, pertinencia y utilidad de la investigación, partiendo del sujeto en relación con los otros, transformándose a sí mismo y a los demás, pero sobre todo, donde surgen nuevas posturas que rompen el silencio, el aquietamiento y la tranquilidad; permitiéndonos a través de nuevos códigos, rebelarnos.

Alterations in matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) have been implicated in adverse left ventricular (LV) remodeling after myocardial infarction (MI). However, the direct mechanistic role of TIMPs in the post-MI remodeling process has not been completely established. The goal of this project was to define the effects of altering endogenous MMP inhibitory control through combined genetic and pharmacological approaches on post-MI remodeling in mice. This study examined the effects of MMP inhibition (MMPi) with PD-166793 (30 mg·kg−1·day−1) on LV geometry and function (conductance volumetry) after MI in wild-type (WT) mice and mice deficient in the TIMP-1 gene [TIMP-1 knockout (TIMP1-KO)]. At 3 days after MI (coronary ligation), mice were randomized into four groups: WT-MI/MMPi ( n = 10), TIMP1-KO-MI/MMPi ( n = 10), WT-MI ( n = 22), and TIMP1-KO-MI ( n = 23). LV end-diastolic volume (EDV) and ejection fraction were determined 14 days after MI. Age-matched WT ( n = 20) and TIMP1-KO ( n = 28) mice served as reference controls. LVEDV was similar under control conditions in WT and TIMP1-KO mice (36 ± 2 and 40 ± 2 μl, respectively) but was greater in TIMP1-KO-MI than in WT-MI mice (48 ± 2 vs. 61 ± 5 μl, P < 0.05). LVEDV was reduced from MI-only values in WT-MI/MMPi and TIMP1-KO-MI/MMPi mice (42 ± 2 and 36 ± 2 μl, respectively, P < 0.05) but was reduced to the greatest degree in TIMP1-KO mice ( P < 0.05). LV ejection fraction was reduced in both groups after MI and increased in TIMP1-KO-MI/MMPi, but not in WT-MI/MMPi, mice. These unique results demonstrated that myocardial TIMP-1 plays a regulatory role in post-MI remodeling and that the accelerated myocardial remodeling induced by TIMP-1 gene deletion can be pharmacologically “rescued” by MMP inhibition. These results define the importance of local endogenous control of MMP activity with respect to regulating LV structure and function after MI.

Recent studies have been directed at modulating the heart failure process through inhibition of activated matrix metalloproteinases (MMPs). We hypothesized that a loss of MMP inhibitory control by tissue inhibitor of MMP (TIMP)-1 deficiency alters the course of postinfarction chamber remodeling and induced chronic myocardial infarction (MI) in wild-type (WT) and TIMP-1−/− mice. Left ventricular (LV) pressure-volume loops obtained from WT and TIMP-1−/− mice demonstrated that LV end-diastolic volume [52 ± 4 (WT) vs. 71 ± 6 (TIMP-1−/−) μl] and LV end-diastolic pressure [9.0 ± 1.2 (WT) vs. 12.7 ± 1.4 (TIMP-1−/−) mmHg] were significantly increased in the TIMP-1−/− mice 2 wk after MI. LV contractility was reduced to a similar degree in the WT and TIMP-1−/− groups after MI, as indicated by a significant fall in the LV end-systolic pressure-volume relationship. Ventricular weight and cross-sectional areas of LV myocytes were significantly increased in TIMP-1−/− mice, indicating that the hypertrophic response was more pronounced. The observed significant loss of fibrillar collagen in the TIMP-1−/−controls may have been an important contributory factor for the observed LV alterations in the TIMP-1−/− mice after MI. These findings demonstrate that TIMP-1 deficiency amplifies adverse LV remodeling after MI in mice and emphasizes the importance of local endogenous control of cardiac MMP activity by TIMP-1.

Background Callitropsis guadalupensis (Guadalupe cypress) is endemic to Guadalupe Island, Mexico, where it is the dominant species of the only forest. The species has suffered declining numbers following the introduction of goats to the island over 150 years ago. Callitropsis guadalupensis is closely related to Callitropsis forbesii (Tecate cypress), distributed in small isolated populations in mainland Baja California and southern California. The objective of the present study was to compare the genetic diversity of the island endemic to the continental species. Methodology/Principal Findings We measured genetic diversity in Callitropsis guadalupensis (n = 54) from Guadalupe Island and in Callitropsis forbesii (n = 100) from five populations in mainland Baja California. The plastid DNA trnS-trnG spacer and the trnL-trnF region were chosen for characterization. Thirty-four haplotypes were observed, of which six were shared between both species. One of these haplotypes was also shared with three other species, Callitropsis lusitanica, Callitropsis montana, and Callitropsis stephensonii. Haplotype diversity (h) and nucleotide diversity (π) were significantly higher for Callitropsis guadalupensis (h = 0.698, π = 0.00071) than for Callitropsis forbesii (h = 0.337, π = 0.00024). Conclusions/Significance Callitropsis guadalupensis shows no evidence of a founder effect or of a genetic bottleneck, and can be added to a growing list of insular species with higher genetic diversity than their mainland relatives.

O Trypanosoma cruzi e considerado um dos eucariotos mais resistentes a Radiação ionizante, suportando doses sub-letais de ate 1,0 kGy. A radiação ionizante do tipo gama promove a quebra das fitas simples e duplas do DNA, além de gerar espécies reativas de oxigênio (ROS). Apos a irradiação, os parasitos cessam o seu crescimento por ate 96 horas, retomando-o em seguida e atingindo a fase estacionaria em ate 240 horas apos irradiação. O padrão de bandas cromossomais, que são extensamente fragmentadas pela radiação, e restabelecido cerca de 48 horas apos o estresse. Para tolerar tais danos, e necessário um eficiente aparato de reconhecimento e reparo de lesões. Neste trabalho, investigamos a radioresistencia em T.cruzi, mediante a análise do seu perfil de expressão protéica e da sua atividade mitocondrial em diferentes tempos apos uma dose de 500 Gy de radiação gama. Utilizando a metodologia 2D-DIGE, foram encontrados 543 spots protéicos diferencialmente expressos apos a exposição do T. cruzi a radiação. A partir destes spots, 53 proteínas foram identificadas por espectrometria de massas (MS/MS) e classificadas de acordo com sua função biológica. Analisando a anotação funcional das 53 proteínas, observamos uma tendência para a superexpressão de proteínas com peso molecular abaixo do predito. Esta tendência indica a possível existência de modificações pos-traducionais e/ou de mecanismos de processamento das proteínas identificadas, que geraria polipeptídios mais curtos do que o predito em resposta a radiação gama. Outras tendências observadas em nossas analises de proteômica incluem alterações nos seguintes processos biológicos: regulação positiva do processo de síntese de proteínas, repressão de proteínas envolvidas no processo de enovelamento (exceto pela indução de duas chaperonas do retículo endoplasmático), repressão das vias de geração de ATP por fosforilação oxidativa, da glicolise, e do metabolismo de aminoácidos. Nossos resultados também indicam que a síntese protéica de#novo e essencial para a recuperação do T. cruzi apos o estresse por exposição a radiação gama, pois o tratamento com inibidores de tradução interfere drasticamente no retorno ao crescimento dos parasitos irradiados. Ao avaliarmos a atividade mitocondrial de T. cruzi, xvi! verificamos que parasitos irradiados produzem mais ATP do que parasitos não irradiados, alem de consumirem mais O2. Adicionalmente, a análise da expressão gênica de T. cruzi irradiado por qRT-PCR mostrou que os genes do maxicírculo mitocondrial foram induzidos apos a irradiação. Ambos os resultados indicam uma maior atividade da mitocôndria apos o estresse. Entretanto, observamos uma menor produção de H2O2 nos parasitos irradiados, indicando assim a existência de um mecanismo eficiente de eliminação de radicais livres. O presente estudo revela a resposta peculiar de T. cruzi a radiação ionizante e suscita questionamentos pertinentes sobre como este organismo e capaz de modificar seu perfil de expressão protéica e sua atividade mitocondrial para possibilitar a sobrevivência em condições tão adversas. The Trypanosoma cruzi is considered as one of the eukaryotes most resistant to ionizing radiation, enduring sub-lethal doses of up to 1.0 kGy. The ionizing radiation by gamma rays promotes double-strand and single-strand breaks on the DNA and, additionally, generates reactive oxygen species (ROS). After irradiation the parasites cease growing for as long as 96 hours, further resuming growth and reaching the stationary phase no later than 240 hours after irradiation. The pattern of chromosomal bands which are extremely fragmented by radiation exposure is restored approximately 48 hours after the stress. To enable the parasite to tolerate such damage condition, an efficient apparatus to recognize and repair lesions is necessary. In the present work we investigate T. cruzi radioresistance by analyzing its protein expression patterns and mitochondrial activity in different time points after a dose of 500 Gy of gamma rays. Using the 2DTDIGE methodology, 543 spots were identified as significantly differentially expressed after T. cruzi exposure to radiation. From these 543 spots, 53 different proteins were identified by mass spectrometry (MS/MS) and classified according to biological function. When the functional annotation of these 53 proteins was analyzed, a tendency of overexpression of proteins with molecular weight lower than predicted was observed. This tendency suggests the presence of post-translational modifications and/or processing mechanisms acting upon the identified proteins, leading to the emergence of peptides which are shorter than predicted in response to gamma radiation exposure. Other tendencies observed on proteomic analysis include alterations on the following biological processes: positive regulation of the protein synthesis process, repression of proteins involved with folding processes (except for two endoplasmic reticulum chaperones, which are induced), repression of ATP generation through oxidative phosphorylation, of glycolysis, and of the amino acids metabolism. Our results also indicate that the de novo protein synthesis is essential for T. cruzi recovery from gamma radiation exposure, since the treatment with translation inhibitors drastically interferes with the growth resumption on irradiated parasites. When the mitochondrial activity of T. cruzi xviii was evaluated, we verified that irradiated parasites generate higher levels of ATP than their non-irradiated counterparts, with an associated higher O2 consumption by the former. Additionally, the gene expression analysis of irradiated T. cruzi using qRT-PCR showed that genes from the mitochondrial maxicircle were induced after radiation exposure. Both of these results point to a higher mitochondrial activity after irradiation-caused stress. Nevertheless, we have also observed a lower H2O2 production by irradiated parasites, thus indicating the existence of an efficient mechanism for free radicals clearance in the organism. The present study unravels the peculiar response of T. cruzi to ionizing radiation and evokes questions about the manner in which this organism is capable of performing drastic changes on its protein expression profile and mitochondrial activity to enable its survival in such adverse conditions.

Brucella abortus é considerada o principal agente etiológico da brucelose bovina, uma zoonose que provoca grandes perdas econômicas em todo o mundo. O objetivo desse estudo foi determinar o perfil diferencial de expressão proteica de células trofoblásticas bovinas nas fases iniciais da infecção por B. abortus. Foi usado o modelo de explantes confeccionados a partir de membrana cório-alantóidea de fetos no último trimestre de gestação infectados por suspensão deB. abortus 2308 (1.0 x 108 bactérias - MOI 1:1000). No estudo da cinética da infecção foram avaliados os tempos de 0,5, 2, 4 e 8 h pós-infecção no qual foi obtida reprodutibilidade alta entre triplicatas dos géis comparados (Match > 84%; IC > 0,77). O perfil de expressão deproteínas foi similar entre os tempos avaliados (Match > 75%). Os tempos de 0,5 e 4 h pósinfecção apresentaram as maiores diferenças qualitativas entre os géis e foram escolhidos para análise diferencial em gel (DIGE). Nessa análise não foram identificados spots com diferençasquantitativas significativas entre os géis comparados. Por outro lado, 103 spots presentes apenas em um grupo experimental foram selecionados para identificação por espectrometria de massa. As proteínas BLVRA, LGALS7, RPLP1, SCAMP2, TOLLIP, GALM, AHCY, PSAPL1/PSAP, OAT, C1QBP, NDUFS8, PRCII, RAB11A, CALM1, MDH1, PRDX3, ABHD14B, KRT14, KRT7, HMGB1, HEXB, AKR1B1, PDIA3 e TPM4 foram identificadas apenas nos extratos proteicos de células trofoblásticas infectadas. As proteínas identificadas nesse estudo servirão como alvos para novas pesquisas relacionadas à interação patógeno-hospedeiro na infecção por B. abortus. Brucella abortus is considered the main etiological agent of bovine brucellosis, a zoonotic disease that causes great economic losses worldwide. The aim of this study was to determine the differential profile of proteins expression of bovine trophoblast cells in the early stages ofinfection by B. abortus. Was used the explants model prepared from chorio-alantóidea of fetuses in the last trimester of pregnancy infected by suspending the B. abortus 2308 containing 1.0 x 108 bacteria (MOI 1:1000). In the study of the kinetics of infection was evaluated the timesof 0.5, 2, 4 and 8 h post infection with B. abortus. Was obtained high reproducibility between triplicate of gels compared (Match> 84%; CI > 0.77) and the proteins expression profile was similar among the times evaluated (Match > 75%). Times of 0.5 and 4 h post infection showedthe highest qualitative differences between the gels and were used for the differential in gel electrophoresis (DIGE). In this analysis have not been identified spots with significant quantitative differences between the compared gels. Moreover, 103 spots are present only in an experimental group were selected for identification by mass spectrometry. The proteins BLVRA, LGALS7, RPLP1, SCAMP2, TOLLIP, GALM, AHCY, PSAPL1/PSAP, OAT, C1QBP, NDUFS8, PRCII, RAB11A, CALM1, MDH1, PRDX3, ABHD14B, KRT14, KRT7, HMGB1, HEXB, AKR1B1, PDIA3 and TPM4 were identified only in protein extracts of trophoblastic cells infected by B. abortus. The proteins identified in this study will serve as targets for further research related to host-pathogen interaction in infection by B. abortus.

A leishmaniose visceral (LV) é uma doença infecciosa grave causada por protozoários do gênero Leishmania. Nas Américas, o cão é o principal reservatório da doença, principalmente nas áreas urbanas, onde encontra-se em grande expansão. A LV é fatal se não for tratada adequadamente e/ou diagnosticada precocemente, porém, a eficácia dos métodos diagnósticos e vacinas disponíveis permanece parcial, sendo necessário o desenvolvimento de novas estratégias. Vários estudos demonstraram o potencial da proteína A2, específica do estágio amastigota de Leishmania, como antígeno vacinal e diagnóstico. No presente trabalho, inicialmente, testamos este antígeno, administrado na forma de vacina de DNA e observamos proteção em camundongos contra a infecção por L. (L.) chagasi e L. (L.) amazonensis. A seguir, descrevemos a proteção, em estudos de fase II, com cães beagles vacinados com a proteína A2 associada ao adjuvante saponina (constituindo a vacina Leish-tec.) contra a infecção por L. (L.) chagasi. Com relação ao diagnóstico utilizando este antígeno, observamos que peptídeos provenientes de análises in silico da proteína A2, associados a peptídeos de outras proteínas como NH, LACK e K39 apresentaram elevada sensibilidade e especificidade em testes com soros humanos ecaninos. No entanto, a busca por antígenos vacinais e diagnóstico na LV é incessante, e o atual estágio de desenvolvimento tecnológico com a possibilidade de utilizar abordagem proteômica para identificação de centenas de antígenos imunogênicos simultaneamente é atraente. Dessa forma, optamos por uma abordagem proteômica associada a análises in silico para identificar novos antígenos de L. (L.) chagasi. Nossa análise proteômica foi a primeira a utilizar amastigotas purificadas de baço de hamster e, além disso, é o primeiro estudo a mapear a antigenicidade da infecção por L. (L.) chagasi utilizando soros de cães de fase aguda e crônica. Todas as proteínas selecionadas porwestern blot-2DE (WB-2DE) foram mapeadas para epítopos de células B pelo software BepiPred e submetidas `a imuno-ensaios em membranas de celulose resultando na seleção de 25 peptídeos com elevado potencial para o diagnóstico sorológico da LV. Paralelamente, foi utilizada a técnica de DIGE para comparar a expressão diferencial deproteínas entre as formas amastigota e promastigota de L. (L.) chagasi e 40 proteínas foram identificadas, sendo 25 aumentadas em promastigotas, 5 aumentadas em amastigotas e 10 com expressão similar em ambas as formas. Além disso, todas as proteínas identificadas no trabalho, tanto por DIGE como por WB-2DE, forammapeadas para epítopos de células T utilizando o software NetCTL que permitiu a seleção de potenciais candidatos para o desenvolvimento de vacinas. Por fim, para complementar a análise de predição para células B, as mesmas proteínas imunogênicas identificadas por WB-2DE forma mapeadas utilizando outros dois programas de predição de epítopos (ABCPred e BCPred) e após análise adicional de imuno-ensaios mais 23 peptídeos foram identificados como potenciais antígenos diagnóstico na LVC. Esses novos peptídeos, juntamente com os 25 previamente identificados, foram submetidos a ensaios imuno-enzimáticos que resultaram em 10 antígenos com elevada sensibilidade e especificidade, inclusive maior do que a encontrada pelo kit EIE-LVC, que é o recomendado pelo Ministério da Saúde, que também falhou em detectar cães assintomáticos. Portanto, uma análise em grande escala de L. (L.) chagasi através do proteoma foi realizada para identificar epítopos de células B e T com potencial para utilização no desenvolvimento de testes diagnósticos e vacinas. Visceral leishmaniasis (VL) is a severe infectious disease caused by a protozoan of the genus Leishmania. In the Americas, the dog is the main reservoir of disease, mainly in urban areas where the disease is increasing. VL is fatal if it is not adequately treated and/or if it is not diagnosed early, however the efficacy of diagnostic methods andvaccines available remain partial, therefore it is necessary to develop new strategies. Several studies have demonstrated the potential of A2 protein, an amastigote stagespecific protein family in Leishmania, as vaccine and diagnosis. In our studies, using this antigen administered as DNA vaccine, we observed protection in mice against infection by L. (L.) chagasi and L. (L.) amazonensis. A2 antigen also induces protectionin phase II studies with beagle dogs vaccinated with A2 protein associated with the saponin adjuvant (constituting the vaccine Leish-tec.) against infection by L. (L.) chagasi. The partial protection induced by A2 antigen in vaccine studies led us to search for new proteins for further vaccine tests and future associations with the A2 antigen. Regarding the diagnosis, we found that the peptides derived from in silico analysis of the A2 protein, associated with peptides from other proteins such as NH, LACK and K39 showed high sensitivity and specificity in tests with human and canine sera. However, the search for vaccine and diagnosis antigens of VL is incessant, and the current stage of technological development with the possibility of using proteomic approach to identify hundreds of immunogenic antigens simultaneously is attractive. Thus, we chose aproteomic approach combined with in silico analyzes to identify new antigens of L. (L.) chagasi. Our proteomic analysis were the first to use amastigotes purified from hamster spleen and, furthermore, is the first study to map the antigenicity of infection by L. (L.) chagasi using sera from dogs of acute and chronic phase. All the selected protein bywestern blot-2DE (WB-2DE) were mapped to B cell epitopes by the BepiPred software and subjected to immunoassay through cellulose membranes resulting in the selection of 25 peptide with high potential for serological diagnosis of VL. In parallel, we used the technique of DIGE to compare the differential expression of proteins between the amastigote and promastigote forms of L. (L.) chagasi and 40 proteins were identified, 25 increased in promastigotes, 5 increased in amastigotes and 10 with similar expression in both forms. Moreover, all the proteins identified in the work by both DIGE as by WB 2DE have been mapped to T cell epitopes using the software NetCTL allowing the selection of potential candidates for vaccine development. Finally, to complete the analysis prediction for B cells, the same immunogenic proteins identified by WB-2DE were mapped using others two programs to predict epitopes (ABCPred and BCPred) and after additional analysis of immunoassays, more 23 peptides were identified as potential diagnostic antigens in the LVC. These new peptides, together with 25 previously identified, were subjected to immuno-enzymatic assays that resulted in 10 antigens withhigh sensitivity and specificity, even greater than that found by the EIE-LVC kit, which is recommended by the Ministry of Health, which also failed to detect asymptomatic dogs. Therefore, a large-scale analysis of L. (L.) chagasi through proteome was performed to identify B and T cell epitopes potentially useful in developing diagnostic tests and vaccines.