ABSTRACT. Gallbladder carcinoma is an uncommon, but highly malignant tumor, with poor prognostic, and diagnostic manifestations in early stages. The Indian Council of Medical Research reported increased incidence of gallbladder carcinoma in the surviving population of the Bhopal gas tragedy that involved exposure of more than 500,000 people to methyl isocyanate gas. The severity of exposure, and increased multi-systemic morbidity in the survivors stimulated us to examine the molecular changes leading to gallbladder carcinoma. Surgically resected samples (N = 40) of gallbladder carcinoma were studied for the p53, Rad50, and cyclin-E expression by immunohistofluorescence bioimaging. Among the 40 samples, 23, 11, and 10 showed p53, Rad50, and cyclin-E expression, respectively, in moderately differentiated adenocarcinomas, demonstrating the prevalence and invasiveness of this disease in the methyl isocyanate-exposed population (P = 0.0009). Nevertheless, co-expression of Rad50, and cyclin-E with p53 was absent in adenomas with dysplasia, demonstrating their independent roles. We conclude that there was altered expression of p53, Rad50, and cyclin-E in the malignant transformation of gallbladder carcinoma in this methyl isocyanate gas-exposed cohort. Hence, these proteins may be useful as markers to identify premalignant lesions that are likely to progress into malignant adenocarcinoma.

ABSTRACT. Epithelioid sarcoma is a rare, aggressive soft tissue tumor of unknown histogenesis showing predominantly epithelioid cytomorphology. We conducted a conventional and molecular cytogenetic study of a 27-year-old male with epithelioid sarcoma with angiomatoid features. Cytogenetic analysis of epithelioid sarcoma metaphase spreads by GTG-banding revealed a diploid chromosome complement with structural and numerical aberrations. Comparative genomic hybridization analysis demonstrated the amplification of 3p24~pter, 4p15.2-p16 and 18q23, while chromosome losses involved 3p13-p14, 3q24-q26.1, 9q21, and 11q21. Fluorescence in situ hybridization assessment showed normal hybridization patterns for the C-MYC and CCND1 loci; CCND1 RNA overexpression was detected by real-time polymerase chain reaction analysis. Genetic evaluation of this rare condition may be useful in determining if epithelioid sarcoma is associated with a distinct genetic background.

ABSTRACT. In the chironomid Acricotopus lucidus, two cells with quite different chromosome complements arise from the last unequal spermatogonial mitosis, as a consequence of monopolar migration of the so-called germ line limited chromosomes (Ks). The cell receiving all the Ks, in addition to two sets of the regularly segregating somatic chromosomes (Ss), develops into the primary spermatocyte, while the cell getting only Ss differentiates into an aberrant spermatocyte. Only the primary spermatocyte enters meiosis. These nuclear events in the primary spermatocytes of A. lucidus during prophase I were analyzed by carmine-orcein staining, silver impregnation, live-cell RNA fluorescence labeling, and fluorescence in situ hybridization, using painting probes of the three Ss and the K centromeres. Early prophase I nuclei display large condensed chromatin blocks showing intense carmine staining, dark silver nitrate impregnation and bright DAPI fluorescence. The first clear signs of meiotic prophase progression are loops arising at early pachytene, which originate from the gradually decondensing chromatin blocks. The blocks presumably represent facultative heterochromatin. Chromosome painting demonstrates that the pachytene loops are composed of the two closely paired homologues. Conspicuous telomere attachments of differently painted non-homologous chromosomes were detected. The centromeres of the Ks group together, indicating a classical bouquet arrangement of the paired homologues in pachytene. The clustered centromeres may function as pairing centers to initiate synapsis of the homologues. Nucleolus expression data support the idea that the aberrant spermatocyte nourishes the primary spermatocyte via a connecting cytoplasmic canal.

ABSTRACT. Down syndrome has been linked to premature aging and genomic instability. We examined the frequency of micronucleus (MN) and binucleated cells in the oral mucosa of Down syndrome patients and healthy controls matched by age and gender, addressing the effect of age and family income. Down syndrome individuals had an increased number of MN (14.30 ± 9.35 vs 4.03 ± 1.71; P 0.001) and binucleated cells (0.97 ± 1.3 vs 0.33 ± 0.66; P 0.05) per 2000 cells. Micronucleus frequency of Down syndrome individuals correlated positively with age (r = 0.437; P = 0.009), and the older (≥21) Down syndrome age group (30.8 ± 8.4 years old) had about 2-fold more micronuclei (P ≤ 0.05) than did the younger group (21). Average family income did not correlate with MN frequency in controls (r = -0.948; P = 0.183), but a borderline negative correlation was seen in DS subjects (r = -0.9484; P = 0.0516). Individuals whose average income was ten times minimum wages had about 2-fold less MN than those receiving around minimum wage. We conclude that the buccal MN assay is a useful and minimally invasive method for monitoring genetic damage in humans and could be used as a tool to evaluate age-associated genomic instability in Down syndrome.

ABSTRACT. Prunus spinosa, blackthorn, exists as wild populations that inhabit uncultivated uplands of Coruh Valley in the northeastern part of Turkey; the fruit is used to make preserves. We examined genetic diversity in wild-grown Prunus spinosa; 16 individual plants from wild populations of Coruh Valley were sampled and subjected to RAPD (random amplified polymorphic DNA) analysis. We tested 51 random decamer primers; 15 of them gave reproducible polymorphic patterns. These 15 primers produced 226 bands, of which 65% were polymorphic. A UPGMA dendrogram clearly divided the genotypes into four groups; we concluded that RAPD analysis can be used for examining genetic relatedness among blackthorn genotypes.

ABSTRACT. The superiority of Africanized over European honey bees in tropical and subtropical regions of the New World is both well documented and poorly understood. As part of an effort to try to understand the process by which the displacement of European bees occurred, we examined the ability of these two types of bees and of hybrids between the two to convert natural and artificial diets into usable protein. Newly emerged bees from colonies of tropically adapted Africanized and temperate-origin Carniolan bees and first-generation hybrids between the two were caged and fed artificial and natural protein diets for six days to determine whether there were differences in their ability to use these diets. The Africanized bees developed significantly higher protein levels in the hemolymph than did the Carniolan bees. The difference was 31% when the bees were fed bee bread (37.5 and 28.56 μg protein/μL hemolymph, re­spectively). The hybrids developed protein levels intermediate be­tween the two parental types. These were approximately 10 times the levels found in bees fed with sucrose alone. Superior food con­version efficiency of the Africanized bees may be one of the reasons for their superiority both in the wild and for beekeeping in tropical and subtropical Latin America.

ABSTRACT. The response of plants to biotic and abiotic stress factors involves biochemical, physiological, morphological, and developmental changes. Salt stress has been the subject of extensive studies due to the low salt tolerance of many crop plants. Germin and germin-like gene products are known to be involved in various aspects of plant development, such as defense, embryonic development and response to biotic as well as abiotic stress, including salt. The responses of germin and germin-like genes to salt stress vary in different plants. RT-PCR and in situ RNA hybridization methods were employed to analyze quantitative changes and to detect the localization pattern of germin gene products in salt-stressed and water-grown wheat embryos. Salt stress caused no quantitative changes in the synthesis of germin gene products. However, in situ RNA hybridization revealed changes in the expression site of germin gene products. Coleorhiza cells were found to be the site of germin accumulation in water-grown embryos; however, in salt-stressed embryos, germin mRNA accumulated in coleoptile cells instead of coleorhiza tissue.

ABSTRACT. Disruption or loss of tumor suppressor gene TP53 is implicated in the development or progression of almost all different types of human malignancies. Other members of the p53 family have been identified. One member, p73, not only shares a high degree of similarity with p53 in its primary sequence, but also has similar functions. Like p53, p73 can bind to DNA and activate transcription. Using PCR-SSCP and gene sequencing, we analyzed the TP53 and TP73 genes in a case of a grade III anaplastic astrocytoma that progressed to glioblastoma. We found a deletion of AAG at position 595-597 of TP53 (exon 6), resulting in the deletion of Glu 199 in the protein and a genomic polymorphism of TP73, identified as an A-to-G change, at position E8/+15 at intron 8 (IVS8-15A>G). The mutation found at exon 6 of the gene TP53 could be associated with the rapid tumoral progression found in this case, since the mutated p53 may inactivate the wild-type p53 and the p73a protein, which was conserved here, leading to an increase in cellular instability.

ABSTRACT. The southern Marmara region in Turkey was surveyed to determine the olive cultivars that are used for olive production. Genetic diversity analysis using simple sequence repeat (SSR) markers indicated that the cultivar Gemlik is the major olive cultivar grown in this region, while other olive cultivars are grown only for use as pollinators of Gemlik or for growers’ own exotic consumption. Although the quality of Gemlik is widely accepted in Turkey, its tendency toward alternate bearing is a major drawback. Twenty-four genotypes were selected within the cultivar Gemlik because of their tolerance to alternate bearing. These selected genotypes have the same SSR alleles as Gemlik, making them good candidates for developing a Gemlik olive with reduced alternate bearing. About 8% of samples did not share the same SSR alleles with Gemlik, though these genotypes were identified as Gemlik by the growers. Some other standard cultivars that are grown in other regions of Turkey were mistakenly called Gemlik in this region, probably due to the popularity of this cultivar in the southern Marmara region. In conclusion, as indicated by SSR analysis, Gemlik has become the standard cultivar in this region; future research should be focused on techniques to improve the production and quality of table olives and olive oil from this cultivar.

ABSTRACT. Plantago ovata, popularly known as isabgol, has great commercial and medicinal importance due to thin rosy white membranous seed husk. Isabgol seeds and husks have emollient, demulcent and laxative properties. We used both biometric and molecular techniques to assess the genetic variability and relatedness of 80 germplasm accessions of Plantago spp (P. ovata, P. lanceolata, and P. major) collected both from India and abroad. The range of D2 values (2.01-4890.73) indicated a very high degree of divergence among the accessions. Based on the degree of divergence, 80 accessions/genotypes were grouped into seven clusters. Thirty-six accessions were analyzed through RAPD profiling for similarity and genetic distances, using 20 random primers. Intraspecific differences in all three species were smaller [range for P. ovata (2-17%), P. lanceolata (3-15%), P. major (2-11%)] than interspecific diversity. These highly divergent lines could be used to produce superior hybrids.

ABSTRACT. Hereditary hemochromatosis is one of the most common autosomal recessive diseases; it is characterized by excess absorption of iron. Clinically, the major challenge is to diagnose increased iron deposition before irreversible tissue damage has occurred. C282Y and H63D are the main mutations related to hereditary hemochromatosis; these mutations have been reported to be associated with increased risk of developing diabetes mellitus type 2 (DM2). We investigated whether these mutations are associated with increased risk for the development of DM2 in women in Brazil. Seventy-two women with clinical diagnosis of DM2 under treatment with hypoglycemic agents and a control group composed of 72 women with no clinical history of diabetes were studied. The C282Y and H63D mutations were determined by PCR-RFLP. Significant differences were not observed for C282Y and H63D, when we compared diabetic and non-diabetic women. We suggest that mutations C282Y and H63D in the HFE gene are not significant risk factors for the development of DM2 in Brazilian women.

ABSTRACT. We evaluated the potential of genetic distances estimated by microsatellite markers for the prediction of the performance of single-cross maize hybrids. We also examined the potential of molecular markers for the prediction of genotypic values and the applicability of the Monte Carlo method for a correlation of genetic distances and grain yield. Ninety S_0:2 progenies derived from three single-cross hybrids were analyzed. All 90 progenies were genotyped with 25 microsatellite markers, including nine markers linked to quantitative trait loci for grain yield. The genetic similarity datasets were used for constructing additive genetic and dominance matrices that were subsequently used to obtain the best linear unbiased prediction of specific combining ability and general combining ability. The genetic similarities were also correlated with grain yield, specific combining ability and heterosis of the hybrids. Genetic distances had moderate predictive ability for grain yield (0.546), specific combining ability (0.567) and heterosis (0.661). The Monte Carlo simulation was found to be a viable alternative for a correlation of genetic distances and grain yield. The accuracy of genotypic values using molecular data information was slightly higher than if no such information was incorporated. The estimation of the relationship using molecular markers proved to be a promising method for predicting genetic values using mixed linear models, especially when information about pedigree is unavailable.

ABSTRACT. We evaluated the phenotypic and genotypic stability and adaptability of hybrids using the additive main effect and multiplicative interaction (AMMI) and genotype x genotype-environment interaction (GGE) biplot models. Starting with 10 single-cross hybrids, a complete diallel was done, resulting in 45 double-cross hybrids that were appraised in 15 locations in Southeast, Center-West and Northeast Brazil. In most cases, when the effects were considered as random (only G effects or G and GE simultaneously) in AMMI and GGE analysis, the distances between predicted values and observed values were smaller than for AMMI and GGE biplot phenotypic means; the best linear unbiased predictors of G and GE generally showed more accurate predictions in AMMI and GGE analysis. We found the GGE biplot method to be superior to the AMMI 1 graph, due to more retention of GE and G + GE in the graph analysis. However, based on cross-validation results, the GGE biplot was less accurate than the AMMI 1 graph, inferring that the quantity of GE or G + GE retained in the graph analysis alone is not a good parameter for choice of stabilities and adaptabilities when comparing AMMI and GGE analyses.

ABSTRACT. An interspecific hybrid between cassava and Manihot glaziovii acquired an apomixis gene from the parent M. glaziovii. This hybrid was exposed to open pollination during three subsequent generations. Seven sibs and the maternal progenitor of the fourth generation were genotyped using six microsatellite loci previously developed for cassava. All sibs were identical with each other and with their maternal progenitor. Sibs of selfed M. glaziovii proved to be identical when examined with these microsatellite loci. The chromosome complement of the apomictic clone was 2n = 38. We observed multi-embryonic aposporic embryo sacs.

ABSTRACT. Chondroitin-4-sulfotransferase-1(C4ST-1)/carbohydrate sul­fotransferase 11 (CHST11) is a Golgi-bound enzyme involved in the biosyn­thesis of the glycosaminoglycan chondroitin sulfate. The sulfation pattern of chondroitin is tightly regulated during development, injury and disease, with the temporal and spatial expression of chondroitin sulfotransferase genes be­lieved to be a crucial determinant of the fine balance of chondroitin sulfation. We have previously identified mouse C4st-1 as a target gene of ligands of the TGFβ superfamily of growth factors, which could positively regulate C4st-1 expression in a number of cell types. We have also shown that a gene trap loss-of-function mutation in C4st-1 leads to severe skeletal abnormali­ties during mouse embryogenesis. In addition, we described a highly specific temporal and spatial expression pattern of C4st-1 during mouse embryogen­esis. However, the transcriptional regulatory mechanisms that control C4st-1 gene expression remain unexplored. In order to gain knowledge on the transcriptional regulation of C4ST-1, we used a bioinformatical approach to identify conserved putative long-range cis-regulatory modules in a region of 120 kb spanning the 5’ end of the C4ST-1 gene. Luciferase reporter assays in human HEK293T and mouse NmuMG cells identified a functional C4ST-1 promoter, as well as a number of cis-regulatory modules able to positively and negatively regulate C4ST-1 expression. Moreover, we identified TGFβ- responsive regulatory modules that can function in a cell type-specific fash­ion. Taken together, our results identify TGFβ-dependent and -independent cis-regulatory modules of the C4ST-1 gene.

ABSTRACT. Glucose-6-phosphate dehydrogenase (G6PD) deficiency is an enzymopathy in which reduced NADPH concentrations are not maintained, resulting in oxidative damage. We evaluated G6PD activity, oxidative stress levels and Trolox equivalent antioxidant capacity in individuals with the A-(202G>A) mutation for G6PD deficiency. Five hundred and forty-four peripheral blood samples were screened for G6PD deficiency; we also analyzed lipid peroxidation products measured as thiobarbituric acid reactive species and Trolox equivalent antioxidant capacity. Men with the A-(202G>A) mutation had lower G6PD activity than women with the same mutation. Individuals with the A-(202G>A) mutation also differed in mean Trolox equivalent antioxidant capacity values but not for thiobarbituric acid reactive species values. We concluded that A-(202G>A) mutation is associated with reduced G6PD activity and increased Trolox equivalent antioxidant capacity.

ABSTRACT. To differentiate among different types of diabetes is becoming an increasingly challenging task. We investigated whether the patient’s genetic profile is useful to identify the particular type of diabetes, to determine the corresponding hyperglycemia pathogenesis and treat accordingly. Three hundred and thirty-eight diabetic patients, diagnosed according to American Diabetes Association criteria, were recruited from 2004 to 2008 in diabetes health reference centers. We analyzed the major gene for type 1 diabetes susceptibility (HLA DQ/DR). In order to improve our understanding of the pathogenesis of the resulting hyperglycemia and to implement a more adequate treatment for the patients, we reclassified our sample according to the presence or absence of the genetic markers. We found that a higher percentage of people than expected have immunological disease, independent of their phenotype, with a relative risk of 4.62 (95% confidence interval). This methodology allowed us to establish an association between the genotype and its resulting phenotype. We found significant differences; the phenotypic classification did not reflect immunological disease based on genotype. Moreover, when we examined markers, body mass index and age of onset, we found that many people have an intermediate phenotype between type 1 and type 2. This genetic data can help provide an accurate definition of the disease and would therefore provide the physician a better possibility of providing adequate treatment.

ABSTRACT. The association between two single nucleotide polymorphisms (SNPs), T945M and UCP1SNP1, with hot carcass weight (HCW, kg, N = 618), longissimus dorsi muscle area (REA, cm2, N = 633), and backfat thickness (BF, mm, N = 625), measured in Nellore cattle in Brazil, was evaluated. Likelihood ratio tests were used to evaluate reduced (fixed effects of general mean, contemporary group, yearling weight, age at slaughter, and random effect of infinitesimal genetic value) and full model (reduced model effects plus quantitative trait locus effects). Additive and dominance effects were tested for each SNP. Genotypic and gene frequencies were also obtained for the SNPs and a descriptive phenotype analysis was made. Mean values for HCW, REA and BF were equal to 288.13 ± 0.55 kg, 73.14 ± 0.27 cm2, and 4.28 ± 0.07 mm, respectively; the coefficients of variation were 4.74, 9.24, and 42.43%, respectively. Gene frequencies for T945M and UCP1SNP1 were f(C) = 0.89, f(T) = 0.11, f(C) = 0.81, and f(G) = 0.19. The SNP T945M had a genotypic frequency of only three animals for TT genotype. Additive effects were observed for T945M on REA and BF, while UCP1SNP1 affected HCW and BF. Based on the significant additive effects of the SNPs and the gene frequencies that we found, we can expect genetic gains with marker assisted selection.

ABSTRACT. β-carotene (BC), pro-vitamin A, is an efficient antioxidant, effective in the neutralization of oxygen reactive species, which cause serious damage to DNA. Various studies have been conducted on the effectiveness of BC for chemoprevention of cancer and heart disease. Doxorubicin is a chemotherapeutic agent used for cancer treatment that generates free radicals. We examined the effects of BC (1, 2 and 4 mg/mL) on the genotoxicity of doxorubicin (0.125 mg/mL), using the wing spot test in Drosophila melanogaster (somatic mutation and recombination test). The BC alone had no significant effect on the frequency of mutant spots. However, it significantly reduced the number of spots caused by doxorubicin. We concluded that BC is not genotoxic and that it exerts protective effects against the genotoxic action of the chemotherapeutic free-radical generator doxorubicin.

ABSTRACT. Genetic diversity of 20 sugarcane accessions in Pakistan was studied using 21 random amplified polymorphic DNA markers. The mean genetic distance between the cultivars was 39.03%, demonstrating that a large part of the genome is similar among the accessions. This probably arises from a lack of parental diversity, with few clones, which are themselves related, contributing to the parentage of these varieties. Among the varieties, none was found to be totally distinct and divergent from the others. We conclude that the current Pakistan commercial varieties have a limited genetic base and that there is a need to diversify commercial sugarcane lines in Pakistan by introducing new germplasm sources.

ABSTRACT. We studied the karyotype, spermatogenesis and nucleolar activity at spermatogenesis in five species of Heteropera: Hyalymenus sp and Neomegalotomus pallescens, Alydidae; Catorhintha guttula and Hypselonotus fulvus, Coreidae; and Niesthrea sidae, Rhopalidae. They showed a red (Alydidae) or orange (Coreidae and Rhopalidae) membrane covering the testes, which consisted of seven testicular lobes, except in N. pallescens, which had only five. All the species had m-chromosomes, an X0 sex chromosome system and 10 (Hyalymenus sp, N. pallescens, and N. sidae), 16 (H. fulvus) or 22 (C. guttula) autosomes. Similar to the other species described to date, all these species showed holocentric chromosomes, interstitial chiasmata in most autosomes, and autosomes dividing reductionally in the first meiotic division and equationally in the second, while sex chromosomes, divided equationally and reductionally in the first and second meiotic divisions, respectively. In addition, we observed that the sex chromosome is heteropycnotic at prophase and that heteropycnotic chromosomal material is found in the nuclei at spermatogenesis; variation in size, shape and location of the nucleolar material occurs during spermatogenesis, denoting a variable degree of activity in the different stages.

ABSTRACT. Embryos produced by hormonal superstimulation have been used as an in vivo control in most published research on embryo gene expression. However, it is not known if this is the most appropriate control for gene expression profile studies. We compared the expression of GRB-10, IGF-II, IGF-IIR, MnSOD, GPX-4, catalase, BAX, and interferon-τ genes, in embryos produced in vivo by hormonal superovulation (SOV), by in vitro fertilization (IVF) or in vivo without any hormonal stimulus (NOV). GRB-10 was less expressed in NOV than IVF embryos, whereas no differences were found for the other genes. The genes related to stress response were then grouped and compared; the sum of expression of MnSOD, GPX-4, and catalase genes tended to be greater in IVF than NOV embryos. A correlation analysis was performed; we found a distinct behavior for NOV embryos when compared with SOV and IVF in the expression of GRB-10, IGF-II and IGF-IIR genes. However, the behavior of these genes was similar in SOV and IVF embryos. We conclude that ovarian hormonal stimulation can affect embryos by altering gene expression. Although this conclusion was based on investigation of only a few genes, we suggest that SOV embryos should be used with caution as a control in gene expression studies.

ABSTRACT. Endophytic bacteria were isolated from stems of Eucalyptus spp (Eucalyptus citriodora, E. grandis, E. urophylla, E. camaldulensis, E. torelliana, E. pellita, and a hybrid of E. grandis and E. urophylla) cultivated at two sites; they were characterized by RAPD and amplified rDNA restriction analysis (ARDRA). Endophytic bacteria were more frequently isolated from E. grandis and E. pellita. The 76 isolates were identified by 16S rDNA sequencing as Erwinia/Pantoea (45%), Agrobacterium sp (21%), Curtobacterium sp (9%), Brevibacillus sp (8%), Pseudomonas sp (8%), Acinetobacter sp (4%), Burkholderia cepacia (2.6%), and Lactococcus lactis (2.6%). Genetic characterization of these endophytic bacteria isolates showed at least eight ARDRA haplotypes. The genetic diversity of 32 Erwinia/Pantoea and 16 Agrobacterium sp isolates was assessed with the RAPD technique. There was a high level of genetic polymorphism among all the isolates and there was positive correlation between the clusters and the geographic origin of the strains. These endophytic bacteria were further analyzed for in vitro interaction with endophytic fungi from Eucalyptus spp. We found that metabolites secreted by Erwinia/Pantoea and B. cepacia isolates had an inhibitory growth effect on some endophytic fungi, suggesting that these metabolites play a role in bacterial-fungal interactions inside the host plant. Apparently, these bacteria could have an important role in plant development; in the future they may be useful for biological control of diseases and plant growth promotion, as well as for the production of new metabolites and enzymes.

ABSTRACT. The androgen receptor (AR) is a protein encoded by the AR gene, which when mutated may affect spermatogenesis, the process in which spermatozoa are produced; thus, AR mutations could lead to male infertility. We examined exon 1 of the AR gene in men with idiopathic infertility. Blood or semen samples from 111 infertile, oligozoospermic (N = 31), asthenozoospermic (N = 23), teratozoospermic (N = 33), and azoospermic (N = 24) men were analyzed. The extracted DNA was amplified for the exon 1 region of the AR gene. There was a significant correlation between the absence of exon 1 in the AR gene and spermatogenesis defects (P = 0.015). This association was significant in teratozoospermic men (51.5% of the sample). We found that lack of amplification of exon 1 of the AR gene by polymerase chain reaction is associated with morphological defects in the spermogram.

ABSTRACT. We estimated general and specific combining abilities and examined resistance to northern leaf blight (Exserohilum turcicum) and to gray leaf spot (Cercospora zeae-maydis) in a set of nine inbred popcorn lines. These inbreds were crossed in a complete diallel scheme without reciprocals, which produced 36 F₁ hybrids. Two experiments with a square lattice design and three replications were conducted during the 2008/2009 crop season, in Maringá, PR, Brazil. The severity of northern leaf blight and gray leaf spot was assessed under natural infestation conditions. Data were examined by individual and joint analysis of variance. Individual and joint Griffing’s diallel analyses were carried out for adjusted means. General combining ability and specific combining ability were significant (P 0.10) by the F-test for northern leaf blight and gray leaf spot infestation levels. This denotes that additive and non-additive gene effects both contributed to resistance to these diseases, but that the additive gene effects were more important. Among the inbred lines, P₈ and P₉ gave the highest resistance to northern leaf blight, and P₃ and P_4.3 gave the highest resistance to gray leaf spot. The hybrids P_7.4 x P₈ and P_4.3 x P₉ could be exploited by reciprocal recurrent selection to provide genotypes with both northern leaf blight and gray leaf spot resistance. Significant interaction between general combining ability and crop season (P 0.10) denotes the importance of environment, even though the disease levels in the hybrids were quite consistent.

ABSTRACT. Human papillomavirus (HPV) has been extensively studied concerning genomic structure, infection mechanisms, and diversity of types, as well as disease progression stages and development of vaccines. HPV type prevalence can differ in specific populations in different countries, according to ethnicity. This is the first report of an integrated project to evaluate the incidence of HPV types in different regions in Brazil in order to obtain data for vaccine development. Cervical samples were collected from women seen at a public hospital in Pernambuco, Northeast Brazil, for routine evaluation of genital alterations. Selection of the patients was random. There was a strong prevalence of HPV16 and a high incidence of HPV types 31 and 33. These data foster the discussion about the need to evaluate viral prevalence in each geographic region in order to develop targeted vaccine programs.

ABSTRACT. Brachiaria humidicola (Poaceae), originally from Africa, is an economically important pasture plant in tropical South America. An accession of B. humidicola (H038) collected from the wild African savanna (Mbeya, Tanzania) showed irregular microsporogenesis. This meiotic behavior was consistent with an allopolyploid origin. Multivalent chromosome association at diakinesis gave tri- to octavalents, associated with two nucleoli in some cells. Six non-congregated univalents in metaphase I and anaphase I, along with previous lines of evidence for x = 6 in B. humidicola, confirm H038 as a nonaploid accession, 2n = 9x = 54. Asynchrony in the genome during microsporogenesis also corroborated this assumption. Its putative origin could be a cross between two related species with different rhythms in meiosis. The meiotic behavior of this accession reinforces the hypothesis of the existence of a new basic chromosome number (x = 6) for Brachiaria. The use of this accession in the breeding of this important forage grass for the tropics is discussed.

ABSTRACT. We examined the influence of the Arg194Trp, Arg280His, and Arg399Gln polymorphisms of XRCC1 (X-ray repair cross-complementing group 1) on the development of childhood acute lymphoblastic leukemia (ALL) in 120 ALL patients and 120 controls in Mexico. All of them were genotyped for these polymorphisms, using polymerase chain reaction. No significant differences in allele and genotype frequencies for any polymorphism were observed between patients and controls. Estimation of haplotypes showed the eight expected haplotypes (A-H), seven of which were found in both patients and controls; haplotype A (Arg-Arg-Arg) was the most common, whereas haplotypes F and G were absent in patients and controls, respectively. Haplotype B (Trp-Arg-Arg) was found to be associated with an increased risk of ALL (odds ratio (OR) = 1.95, 95% confidence interval (CI) = 1.13-3.37; P = 0.016), particularly in males (OR = 2.65, 95%CI = 1.25-5.63; P = 0.01). Individually, the 194Trp, 280His, and 399Gln alleles were not associated with significantly increased risk for ALL in these Mexican children.

ABSTRACT. Ten maize accessions (NC-9, A50-2, M-14, B-42, NC-3, T-7, N-48-1, B-34, USSR, and WFTMS) were studied to estimate the genetic distance on molecular level by random amplified polymorphic DNA. These accessions were selected on the basis of their variable responses against different levels of moisture. Twenty-five primers were used to test genetic diversity, of which 14 were observed to be polymorphic. Ninety-three loci were amplified; among these, 77 showed polymorphism and the other 16 were monomorphic. Primers A-13 and C-02 gave the most polymorphic bands, while primers A-01 and C-06 gave the fewest polymorphic bands. The genetic similarities of the 10 maize accessions ranged from 82.8 to 54.8%. Accessions USSR and WFTMS showed greatest similarity, and accessions M-14 and B-42 were found more dissimilar than the other accessions. On the basis of cluster analysis, these 10 accessions were classified in two major groups, A and B, and than further divided into sub-groups. The cluster analysis showed that accessions in the same group as well as in the sub-groups were similar in their physical and morphological characters, since the characters are controlled genetically.

ABSTRACT. RBM22 is a newly discovered RNA-binding motif protein, belonging to the SLT11 family; it has been reported to be involved in pre-splicesome assembly and to interact with the Ca2+-signaling protein ALG-2. However, previous studies have not demonstrated whether its expression is essential for early embryogenesis in vertebrates. We utilized zebrafish as a developmental model to study the role of RBM22 during embryogenesis. The aligned protein sequences of RBM22 were found to be highly conserved. In situ analysis showed that zRBM22 was expressed as early as the one-cell stage, implying maternal origin during oogenesis; embryos continued to express zRBM22 through at least the 32-h post-fertilization (hpf) stage of development. RT-PCR analysis confirmed the in situ expression pattern, indicating that RBM22 is expressed without any spatial and temporal specificity during zebrafish development. zRBM22 morpholino-induced alterations in development were observed as early as 15 hpf of embryogenesis, based on a morphological analysis. Embryogenesis was monitored through 32 hpf; knockdown resulted in a truncated axis and developmental arrest of the head and tail. We conclude that RBM22 plays an important role in zebrafish embryogenesis.

ABSTRACT. Bovine papillomaviruses (BPV) are the causal agents of benign and malignant lesions; they can cause dramatic economic losses in cattle. Although 10 virus types have been described, three types are most common in tumors, namely BPV-1, -2 and -4. Previous studies have reported BPV in blood cells and the possibility of blood acting as a latent virus site and/or transmission agent of virus dissemination. We studied a Holstein dairy herd in Pernambuco, Brazil, in which several animals showed severe cutaneous papillomatosis, without previous determination of BPV types. Blood samples and short-term lymphocyte cultures were collected from 54 cows. We compared the BPV types detected in peripheral blood to those identified in the respective lymphocyte cultures: BPV-1 was detected in 74% and BPV-2 in 87% of the whole blood samples. Simultaneous virus presence (BPV-1 and BPV-2) was found in 65% of the blood samples. BPV-1 or BPV-2 were detected in the lymphocyte cultures in 93% of the samples, and both in 89%. The detection of viral DNA in whole blood and in lymphocyte cultures is evidence that this virus is carried by lymphocytes.

ABSTRACT. Physical training induces beneficial adaptations; however, exhausting exercise increases reactive oxygen species generation, resulting in damage to DNA and tissues. Pequi (Caryocar brasiliense), a fruit of the Brazilian Cerrado, contains a carotenoid-rich oil. We investigated whether pequi oil had antioxidant effects in runners. Evaluations were made after outdoor races before and after ingestion of 400 mg pequi-oil capsules for 14 days. Blood samples were taken after races and submitted to comet and TBARS assays and biochemical analyses of creatine kinase (CK), aspartate aminotransferase (AST) and alanine aminotransferase (ALT). To determine if the protective effects of pequi-oil were influenced by antioxidant enzyme genotypes, MnSOD (-Val9Ala), CAT (-21A/T) and GPX1 (Pro198Leu) gene polymorphisms were also investigated. Pequi oil was efficient in reducing tissue injuries evaluated for AST and ALT, particularly in women, and in reducing DNA damages in both sexes. Except for CK levels, the results were influenced by MnSOD genotypes; heterozygous excess was related to less DNA damage, tissue injury and lipid peroxidation, besides presenting a better response to pequi oil against exercise-induced damage.

ABSTRACT. Four species of Aplastodiscus and two species of Hypsiboas were cytogenetically compared. Aplastodiscus perviridis, A. cochranae, H. albomarginaus, and H. faber had 2n = 24 chromosomes, while A. albosignatus and A. leucopygius had 2n = 20 and 2n = 18 chromosomes, respectively. Aplastodiscus perviridis and A. cochranae had identical karyotypes, as indicated by their chromosomal morphology, the location of the nucleolus organizer region (NOR) on chromosome pair 12, and the heterochromatin pattern. The NOR-bearing chromosomes of A. albosignatus and A. leucopygius (pair 9) were very similar in size and morphology (metacentric) when compared to A. perviridis and A. cochranae (pair 12) and to H. faber (pair 11); the NOR of these chromosomes also occurred in the same region, suggesting that these chromosomes are homologous. Although H. albomarginatus differs from the other species with regard to the location of its NOR on pair 2, this species had the same diploid number and a chromosomal morphology similar to that of A. perviridis and A. cochranae. Chromosomal differentiation among the species appears to have occurred by reduction in chromosome number.

ABSTRACT. Mutations of some contributing factors (p53, p16, Rb, and EGFR) are believed to affect diagnosis and drug resistance of lung cancer. We evaluated the efficacy of a multimarker panel for molecular diagnosis of lung cancer, using a high-throughput suspension microarray. One hundred and twenty-five lung cancer specimens and 30 tumor-free lung tissue samples were assayed by multiplex polymerase chain reaction with specific probes designed to detect hot-spot mutations in p53, p16, Rb, and EGFR. The mutation rates of p53, p16, Rb, or EGFR in the lung cancer specimens were 36.8, 15.2, 11.2, and 18.4%, respectively. Inclusion of four markers elevated sensitivity to 68.0%. The specificity and accuracy of four-marker detection were 90.0 and 72.3%, and the mutation rates of this panel in stage I, stage II and stage III disease were 62.2, 65.9 and 75.0%, respectively. Mutation at p16 occurred more frequently in non-small cell lung cancer (19.3%) than in small cell lung cancer (5.4%); while the mutation rate of Rb was 32.4% in small cell lung cancer versus 2.3% in non-small cell lung cancer. We conclude that simultaneous detection of p53, Rb, p16, and EGFR in a suspension microarray facilitates rapid diagnosis of lung cancer.

ABSTRACT. One hundred and fifteen unrelated Mangalarga Marchador horses were sampled from three geographically distinct regions of Minas Gerais State, Brazil (South, Southeast, and Northeast) and tested for 10 microsatellite loci. Genetic diversity and population structure parameters were estimated with ARLEQUIN 3.0, CERVUS 2.0, POPGENE 1.31, GENEPOP on the web, STRUCTURE 2.0, and SPAGEDI 1.2 software packages. Under Hardy-Weinberg assumptions, seven markers were at equilibrium (LEX014, LEX017, LEX019, SGCV23, TKY321, VHL20, and VIASH39), while two (ASB3 and LEX031) presented significant homozygote excess. Seventy-four alleles were identified in these nine markers, with a mean of 8.22 alleles. Mean heterozygosity was 0.637 and polymorphism information content was 0.662. Markers ASB3, LEX019, SGCV23, TKY321, and VHL20 were highly informative (PIC > 0.7) and may be useful for eventual expansion of parentage test panels. The FST value (0.0562) indicated relatively little geographical structure. However, based on a Bayesian-based cluster analysis under a three-cluster model, 94% of the 115 individuals were correctly assigned to the subpopulations from where they were sampled. Mean pairwise f was relatively high (0.11), and in spite of the efforts towards non-consanguineous sampling, 1% of the pairs of individuals shared over 50% of the alleles. These results strongly suggest that the population is genetically structured. Under a conservation genetics approach, two strategies are recommended: avoidance of crosses between highly endogamic individuals and stimulation of crosses between individuals from those regions for which low genetic flow was identified.