Cytogenetic characterization and evaluation of c-MYC gene amplification in PG100, a new Brazilian gastric cancer cell line

Gastric cancer is the fourth most frequent type of cancer and the second cause of cancer mortality worldwide. The genetic alterations described so far for gastric carcinomas include amplifications and mutations of the c-ERBB2, KRAS, MET, TP53, and c-MYC genes. Chromosomal instability described for gastric cancer includes gains and losses of whole chromosomes or parts of them and these events might lead to oncogene overexpression, showing the need for a better understanding of the cytogenetic aspects of this neoplasia. Very few gastric carcinoma cell lines have been isolated. The establishment and characterization of the biological properties of gastric cancer cell lines is a powerful tool to gather information about the evolution of this malignancy, and also to test new therapeutic approaches. The present study characterized cytogenetically PG100, the first commercially available gastric cancer cell line derived from a Brazilian patient who had a gastric adenocarcinoma, using GTG banding and fluorescent in situ hybridization to determine MYC amplification. Twenty metaphases were karyotyped; 19 (95%) of them presented chromosome 8 trisomy, where the MYC gene is located, and 17 (85%) presented a deletion in the 17p region, where the TP53 is located. These are common findings for gastric carcinomas, validating PG100 as an experimental model for this neoplasia. Eighty-six percent of 200 cells analyzed by fluorescent in situ hybridization presented MYC overexpression. Less frequent findings, such as 5p deletions and trisomy 16, open new perspectives for the study of this tumor.

B-type natriuretic peptide in the evaluation of cardiac function

B type natriuretic peptide (BNP) and its precursor, the inactive form of NT-pro-BNP, are currently the most studied laboratory parameters in the heart disease spectrum. The assessment of their blood concentrations provides invaluable information on the likelihood, severity and prognosis of the disease. The present review aims to describe the biological determinants, the factors that influence these peptide concentrations, the suggested cutoff values for the diagnosis of heart failure and the use of this biomarker in the assessment of cardiac function.

The role of B-mode ultrasonography in the musculoskeletal anatomical evaluation of the cervical region of the dog spine

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Molecular Evaluation of Developmental Competence of Oocytes Collected In Vivo from Buffalo and Bovine Heifers during Winter and Summer

Buffaloes and bovines are polyestrous and seasonal or annual livestock, respectively, that show reduced fertility during heat stress. To investigate whether reduced fertility is related to oocyte competence in both species, immature oocytes from buffalo and bovine heifers were collected during winter and summer and subjected to molecular analyses. In each season, heifers of both species had their follicular wave emergence synchronized with a standard protocol (Ferreira et al., 2011). Before being subjected to ovum pick up (OPU), cutaneous (CT; degrees C) and rectal (RT; degrees C) temperatures and respiratory rate (RR; breaths/min) were measured. Oocytes' RNA was extracted to evaluate the expression of target genes related to mtDNA replication/transcription (PPARGC1A, TFAM and MT-CO1), apoptosis (BAX and BCL2) and HS (HSP90AA1 and HSPA1AB). ACTB, HIST1H2AG and GAPDH were initially chosen as housekeeping genes. In buffaloes, CT (35.0 +/- 0.4 vs 23.8 +/- 0.5), RT (38.7 +/- 0.1 vs 38.0 +/- 0) and RR (21.3 +/- 1.2 vs 15.4 +/- 1.1) were higher during summer than winter. However, in bovine heifers, RT (38.7 +/- 0.1 vs 38.6 +/- 0.1) and RR (44.8 +/- 1.5 vs 40.6 +/- 1.5) were similar in both seasons, while CT (31.6 +/- 0.3 vs 30.2 +/- 0.3) was increased during summer. Reduced expression of ACTB, HIST1H2AG and GAPDH was evidenced during summer, disqualifying them as housekeeping genes. Similarly, the expression of all target genes was reduced during summer in oocytes of both species. In summary, physiological responses to heat stress seem to be more intense in buffalo than bovine heifers. However, in both species, negative effects of heat stress upon oocyte quality occur at the molecular level and affects genes related to several biological functions.

Evaluation of sperm quality in successive regular collections from captive black-and-gold howler monkeys (Alouatta caraya)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

EGHEP-ISFG collaborative exercise on mixture profiles of autosomal STRs (GHEP-MIX01, GHEP-MIX02 and GHEP-MIX03): Results and evaluation

One of the main objectives of the Spanish and Portuguese-Speaking Group of the International Society for Forensic Genetics (GHEP-ISFG) is to promote and contribute to the development and dissemination of scientific knowledge in the area of forensic genetics. Due to this fact, GHEP-ISFG holds different working commissions that are set up to develop activities in scientific aspects of general interest. One of them, the Mixture Commission of GHEP-ISFG, has organized annually, since 2009, a collaborative exercise on analysis and interpretation of autosomal short tandem repeat (STR) mixture profiles. Until now, three exercises have been organized (GHEP-MIX01, GHEP-MIX02 and GHEP-MIX03), with 32, 24 and 17 participant laboratories respectively. The exercise aims to give a general vision by addressing, through the proposal of mock cases, aspects related to the edition of mixture profiles and the statistical treatment. The main conclusions obtained from these exercises may be summarized as follows. Firstly, the data show an increased tendency of the laboratories toward validation of DNA mixture profiles analysis following international recommendations (ISO/IEC 17025: 2005). Secondly, the majority of discrepancies are mainly encountered in stutters positions (53.4%, 96.0% and 74.9%, respectively for the three editions). On the other hand, the results submitted reveal the importance of performing duplicate analysis by using different kits in order to reduce errors as much as possible. Regarding the statistical aspect (GHEP-MIX02 and 03), all participants employed the likelihood ratio (LR) parameter to evaluate the statistical compatibility and the formulas employed were quite similar. When the hypotheses to evaluate the LR value were locked by the coordinators (GHEP-MIX02) the results revealed a minor number of discrepancies that were mainly due to clerical reasons. However, the GHEP-MIX03 exercise allowed the participants to freely come up with their own hypotheses to calculate the LR value. In this situation the laboratories reported several options to explain the mock cases proposed and therefore significant differences between the final LR values were obtained. Complete information concerning the background of the criminal case is a critical aspect in order to select the adequate hypotheses to calculate the LR value. Although this should be a task for the judicial court to decide, it is important for the expert to account for the different possibilities and scenarios, and also offer this expertise to the judge. In addition, continuing education in the analysis and interpretation of mixture DNA profiles may also be a priority for the vast majority of forensic laboratories. (C) 2014 Elsevier Ireland Ltd. All rights reserved.

Multiple-trait genomic evaluation for milk yield and milk quality traits using genomic and phenotypic data in buffalo in Brazil

The use of markers distributed all long the genome may increase the accuracy of the predicted additive genetic value of young animals that are candidates to be selected as reproducers. In commercial herds, due to the cost of genotyping, only some animals are genotyped and procedures, divided in two or three steps, are done in order to include these genomic data in genetic evaluation. However, genomic evaluation may be calculated using one unified step that combines phenotypic data, pedigree and genomics. The aim of the study was to compare a multiple-trait model using only pedigree information with another using pedigree and genomic data. In this study, 9,318 lactations from 3061 buffaloes were used, 384 buffaloes were genotyped using a Illumina bovine chip (Illumina Infinium (R) bovineHD BeadChip). Seven traits were analyzed milk yield (MY), fat yield (FY), protein yield (PY), lactose yield (LY), fat percentage (F%), protein percentage (P%) and somatic cell score (SCSt). Two analyses were done: one using phenotypic and pedigree information (matrix A) and in the other using a matrix based in pedigree and genomic information (one step, matrix H). The (co) variance components were estimated using multiple-trait analysis by Bayesian inference method, applying an animal model, through Gibbs sampling. The model included the fixed effects of contemporary groups (herd-year-calving season), number of milking (2 levels), and age of buffalo at calving as (co) variable (quadratic and linear effect). The additive genetic, permanent environmental, and residual effects were included as random effects in the model. The heritability estimates using matrix A were 0.25, 0.22, 0.26, 0.17, 0.37, 0.42 and 0.26 and using matrix H were 0.25, 0.24, 0.26, 0.18, 0.38, 0.46 and 0.26 for MY, FY, PY, LY, % F, % P and SCCt, respectively. The estimates of the additive genetic effect for the traits were similar in both analyses, but the accuracy were bigger using matrix H (superior to 15% for traits studied). The heritability estimates were moderated indicating genetic gain under selection. The use of genomic information in the analyses increases the accuracy. It permits a better estimation of the additive genetic value of the animals.

Learning strategies evaluation in students with attention deficit hyperactivity disorder

This study aim to verify the use of learning strategies in students of the elementary level presenting interdisciplinary diagnosis of attention dei cit hyperactivity disorder (ADHD). Nine students, male gender, attending 3rd to 9th grade level of the elementary level, average age 10 years and 7 months, presenting interdisciplinary diagnosis of attention dei cit hyperactivity disorder (ADHD). h e students were submitted to the application of the Evaluation of Learning Strategies from elementary level – EAVAP-EF – scale, which aimed to evaluate the strategies reported and used by students in situation of study and learning, as follows: cognitive strategies, metacognitive strategies and absence of dysfunctional metacognitive strategies. h e general result at EAVAP-EF scale, showed that students with ADHD reached the percentile 25%, considered as low performance in the use of the learning strategies. For the variable absence of dysfunctional metacognitive strategies, the students presented percentile 30%, percentile 25% for cognitive strategies and 55% for metacognitive strategies. h e results showed that ADHD students do not use ef ectively the learning cognitive and metacognitive strategies and present the use of dysfunctional metacognitive strategies. h ese alterations match with the framework of ADHD because the entry of information, either visual or auditory, showed alterations, derived from inattention, which af ected the learning in classroom situation.

Evaluation of indexed language used in collective catalogues of university libraries: a socio-cognitive study using verbal protocol

The use of indexing language in university libraries collective catalogs and the socio-cognitive context of indexing and users were evaluated. The methodology consisted of a diagnostic study elaboration of the functioning and treatment procedures of the indexing information from nine libraries of the UNESP Network, representing the Civil Engineering, Pedagogy and Dentistry areas from a data collection using the Verbal Protocol introspective technique in the Individual and Group forms. The study conducted a reflection upon the statements issued by the seventy-two participating individuals whose the results revealed unsatisfactory results about the use of the Subject Headings List of the BIBLIODATA Network, indexing language utilizing by the UNESP Libraries Network, Brazil, in the representation and in the information retrieval process in the ATHENA catalog, about the sequent aspects of the language: lack of specialized vocabulary as well as updated; lack of remissives and of specific headings, and others. We have concluded that the adequate use of indexing languages of specialized scientific areas becomes by means of evaluation as to updating, specificity and compatibility in order to meet the needs of indexing and information retrieval.

Preparation, characterization and evaluation of drug-delivery systems: Pectin and mefenamic acid films

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

EVALUATION OF EUCALYPTUS KRAFT PULP LIGNIN THROUGH THE NITROBENZENE OXIDATION, COPPER OXIDE REDUCTION AND ACIDOLYSIS TECHNIQUES

This study aimed to evaluate the sensitiveness of the information obtained for the residual lignin from Eucalyptus grandis kraft pulps analyzed through the nitrobenzene oxidation, copper oxide (CuO) reduction and acidolysis techniques. The chips were cooked, resulting pulps of kappa number 14,5 and 16,9, respectively. Both lignins' pulps were evaluated through three methods (nitrobenzene oxidation, copper oxide oxidation and acidolysis). Then, they were subjected to an oxygen delignification stage. The 16,9 kappa number pulp resulted in higher levels of non-condensed lignin structures by the acidolysis method, higher syringyl/vanillin ratios (S/V) by the nitrobenzene and copper oxide methods and better performance in the oxygen delignification stage. The different methods allowed to differ the residual lignin pulps with kappa number 14,5 and 16,9, and the nitrobenzene oxidation method showed the highest sensitiveness in this study results.

Evaluation of dental restorations: a comparative study between clinical and digital photographic assessments

The aim of this study was to compare the efficacy of a direct clinical evaluation method with an indirect digital photographic method in assessing the quality of dental restorations. Seven parameters (color, occlusal marginal adaptation, anatomy form, roughness, occlusal marginal stain, luster, and secondary caries) were assessed in 89 Class I and Class II restorations from 36 adults using the modified US Public Health Service/Ryge criteria. Standardized photographs of the same restorations were digitally processed by Adobe Photoshop software, separated into the following four groups and assessed by two calibrated examiners: Group A: The original photograph displayed at 100%, without modifications (IMG100); Group B: Formed by images enlarged at 150% (IMG150); Group C: Formed by digital photographs displayed at 100% (mIMG100), with digital modifications (levels adjustment, shadow and highlight correction, color balance, unsharp Mask); and Group D: Formed by enlarged photographs displayed at 150% with modifications (mIMG150), with the same adjustments made to Group C. Photographs were assessed on a calibrated screen (Macbook) by two calibrated clinicians, and the results were statistically analyzed using Wilcoxon tests (SSPS 11.5) at 95% CI. Results: The photographic method produced higher reliability levels than the direct clinical method in all parameters. The evaluation of digital images is more consistent with clinical assessment when restorations present some moderate defect (Bravo) and less consistent when restorations are clinically classified as either satisfactory (Alpha) or in cases of severe defects (Charlie). Conclusion: The digital photographic method is a useful tool for assessing the quality of dental restorations, providing information that goes unnoticed with the visual-tactile clinical examination method. Additionally, when analyzing restorations using the Ryge modified criteria, the digital photographic method reveals a significant increase of defects compared to those clinically observed with the naked eye. Photography by itself, without the need for enlargement or correction, provides more information than clinical examination and can lead to unnecessary overtreatment.

Evaluation of the Atibaia River water quality using Lecane bulla as a test organism

In view of the diversity of environments found in the Brazilian territory, it is understandable that the use of native species can provide more relevant information for ecotoxicological studies. The purpose of this work was to evaluate the quality of water samples from the Atibaia River in an area that is under the influence of petroleum refinery using a native test-organism and submitting the data to PCA statistical analysis. Therefore, acute toxicity assays with Lecane bulla (Rotifera) were performed in four locations of the river, as well as physical-chemical analyses. Sampling was drawn in the dry and rainy seasons. The bioassays were static and lasted 48 hours; dead organisms were quantified at the end of the tests. Toxicological differences among the samples/per location and control were compared by means of the Analysis of Variance. Physical-chemical and mortality variables were simultaneously analyzed by multivariate analysis of the principal components and the Pearson correlation coefficient. Water samples from the exit of the refinery stabilization pond (location S.1) were toxic to L. bulla in both seasons, with significant differences in relation to the control and between the seasons. The statistical treatment of data showed that mortality was strong and positively correlated with total hardness, chlorides and EC, which together with pH presented higher values in location S.1, in the dry and in the rainy seasons. Due to its sensibility to the quality of the Atibaia river water samples, the potential use of L. bulla for ecotoxicological studies as an alternative test organism could be demonstrated.

Bone marrow cytological evaluation in dogs with chronic kidneydisease

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)