Scielo RSS <![CDATA[Genetics and Molecular Biology]]> vol. 30 num. 3 lang. en <![CDATA[SciELO Logo]]> <![CDATA[<B>Matrix metalloproteinase gene polymorphisms in patients with coronary artery disease</B>]]> Matrix metalloproteinases (MMPs) play an important role in the pathogenesis of atherosclerosis, the pathology underlying the majority of coronary artery disease (CAD). In this study we tested the hypothesis that polymorphic variation in the MMP genes influences the risk of developing atherosclerosis. We analyzed functional polymorphisms in the promoter of the MMP-1, MMP-3, MMP-9 and MMP-12 genes in 183 Brazilian Caucasian individuals submitted to coronary angiography, of which 67 (37%) had normal coronary arteries (control group) and 116 (63%) had CAD (CAD patient group). The -1607 1G/2G MMP-1, -1171 5A/6A MMP-3, -1562 C/T MMP-9, -82 A/G MMP-12 polymorphisms were analyzed by PCR followed by restriction digestion. No significant differences were observed in allele frequencies between the CAD patients and controls. Haplotype analysis showed no differences between the CAD patients and controls. There was a significant difference in the severity of CAD, as assessed by the number of diseased vessels, in MMP-1 1G/1G homozygous individuals and in those homozygous for the 6A allele of the MMP-3 polymorphism. However, multivariate analysis showed that diabetes mellitus was the only variable independently associated with CAD severity. Our findings indicated that MMP polymorphisms have no significant impact on the risk and severity of CAD. <![CDATA[<B>Cytochrome P450 (CYP) and glutathione S-transferases (GST) polymorphisms (CYP1A1, CYP1B1, GSTM1, GSTP1 and GSTT1) and urinary levels of 1-hydroxypyrene in Turkish coke oven workers</B>]]> Genetic polymorphisms of xenobiotic metabolizing enzymes have been associated with cancer risk. We evaluated the influences of genetic polymorphisms of polycyclic aromatic hydrocarbon (PAH) metabolizing enzymes on urinary 1-hydroxypyrene (1-OHP) excretion in Turkish coke oven workers. Urinary 1-OHP was analyzed by HPLC after enzymatic hydrolysis. Lymphocyte DNA was used for PCR-based genotyping of cytochrome P450 (CYP) polymorphisms (CYP1A1 and CYP1B1) and glutathione S-transferases (GST) polymorphisms (GSTM1, GSTT1 and GSTP1). The mean urinary 1-OHP levels of coke oven workers were significantly higher than that of controls. No significant difference was detected in the mean urinary 1-OHP levels of smokers and non-smokers either for coke oven workers or controls. Genetic polymorphisms of the CYPs and GSTs studied had no significant influence on 1-OHP excretion in coke oven workers, but in the control group the urinary 1-OHP levels of individuals carrying the GSTT1- genotype were significantly higher than those of individuals carrying GSTT1+ genotype. The duration of occupational exposure and metabolic genotype for GSTT1 were the significant predictors of urinary 1-OHP levels. The control individuals carrying combined GSTM1-/GSTT1- genotypes also had significantly higher levels of urinary 1-OHP than those of individuals carrying GSTM1+/GSTTI+, GSTM1-/GSTT1+, and GSTM1+/GSTT1- genotypes. These results indicate that urinary 1-OHP is a sensitive indicator of recent human exposure to PAHs and that genetic polymorphism of GSTT1 may also to some extent reflect the interindividual variation in susceptibility to PAHs only at low PAH exposure. <![CDATA[<B>An analysis of multiplex-PCR in the detection of BCR-ABL transcripts in hematological disorders</B>]]> In this work, we describe the advantages of multiplex-PCR in the specific detection of BCR-ABL transcripts in different hematological disorders and its sensitivity compared to nested PCR. Fifty-three patients were studied for the presence of BCR-ABL transcripts: 24 patients with chronic myeloid leukemia (CML), 20 with acute leukemia (AL), and 9 patients with other hematological disorders. A variant rearrangement (b3a3) was found in a single case of CML (4.2%). Four out of the 20 patients with AL (20.0%) (14 adults, 6 children) were bcr-abl(+), and in this group three cases were classified as B-acute lymphoblastic leukemia (B-ALL), and one as acute myeloblastic leukemia (AML). Two of the three patients with B-ALL were positive for b2a2 and the other one for e1a2, while in the BCR-ABL(+)AML patients a b3a2 rearrangement was observed. In conclusion, multiplex-PCR allows rapid, specific and simultaneous detection of different types of BCR-ABL transcripts in CML and ABL-BCR(+)AL. A full correlation was observed when multiplex-PCR was compared with nested PCR. <![CDATA[<B>Common N-acetylgalactosamine-6-sulfate sulfatase (<I>GALNS</I>) exon mutations in Brazilian patients with mucopolysaccharidosis IVA (MPS IVA)</B>]]> Morquio A Syndrome (mucopolysaccharidosis IVA - MPS IVA, OMIM# 253000) is an autosomal recessive inborn error of metabolism caused by the deficiency of N-acetylgalactosamine-6-sulfate sulfatase (GALNS). We investigated five unrelated Brazilian MPS IVA families for mutations in exons 4, 5, 9 and 10 of the GALNS gene. Six out of the 10 mutant alleles were identified. Taken together with a previous study, which included six unrelated families, common mutations among Brazilian patients were p.N164T, p.G116S and p.G301C. Among one hundred control subjects three novel silent mutations were found (p.A107A; GCC -> GCT, p.Y108Y; TAC -> TAT, p.P357P; CCG -> CCA). Screening starting with exons 4, 5, 9, 10 and 11 may be a good strategy for genotyping of Brazilian patients since these exons include 73% of all mutations identified in the current and previous studies. <![CDATA[<B>Satellite DNA sites in four species of the genus <I>Astyanax</I> (Teleostei, Characiformes)</B>]]> Cytogenetic data about satellite DNA distribution in four Astyanax species (Characidae) from the Paraitinga river, Paraíba do Sul river basin, Brazil, are presented. In order to characterize the constitutive heterochromatin, C-banding, chromomycin A3 and DAPI fluorescence staining, as well as fluorescence in situ hybridization (FISH) with the satellite DNA As-51 probe were performed. A. scabripinnis and A. parahybae presented 2n = 50 and 2n = 48 chromosomes, respectively. The heterochromatin was located in the pericentromeric and terminal regions of many chromosomes, corresponding to GC-positive regions and to the As-51 satellite DNA in terminal regions. A. intermedius and A. giton, both with 2n = 50 chromosomes, showed little heterochromatin, mostly restricted to the terminal and pericentromeric regions of a few chromosomes. No GC-positive regions, neither any correspondence between the scarce heterochromatin of these species and the As-51 satellite DNA was observed. AT-positive blocks were not detected in any of the species studied. Based on these and other available data, the hypothesis that Astyanax represents a polyphyletic group is discussed. <![CDATA[<B>Use of ridge regression for the prediction of early growth performance in crossbred calves</B>]]> The problem of multicollinearity in regression analysis was studied. Ridge regression (RR) techniques were used to estimate parameters affecting the performance of crossbred calves raised in tropical and subtropical regions by a model including additive, dominance, joint additive or "profit heterosis" and epistatic effects and their interactions with latitude in an attempt to model genotype by environment interactions. A software was developed in Fortran 77 to perform five variant types of RR: the originally proposed method; the method implemented by SAS; and three methods of weighting the RR parameter lambda. Three mathematical criteria were tested with the aim of choosing a value for the lambda coefficient: the sum and the harmonic mean of the absolute Student t-values and the value of lambda at which all variance inflation factors (VIF) became lower than 300. Prediction surfaces obtained from estimated coefficients were used to compare the five methods and three criteria. It was concluded that RR could be a good alternative to overcome multicollinearity problems. For all the methods tested, acceptable prediction surfaces could be obtained when the VIF criterion was employed. This mathematical criterion is thus recommended as an auxiliary tool for choosing lambda. <![CDATA[<B>Bayesian inference in a quantitative genetic study of growth traits in Nelore cattle (<I>Bos indicus</I>)</B>]]> The objective of this study was to estimate (co)variance components and genetic parameters for weights (W) and scrotal circumferences (SC) at 365 and 450 days of age, of Nelore (Bos indicus) cattle, using Bayesian inference in single and multiple-trait animal models. The fitted linear models included, besides the animal and residual random effects, the contemporary group (herd-year-season-sex-management group) and age-of-dam as "fixed effects". The analyses were carried out using a Gibbs sampler implemented through the MTGSAM program. The heritabilities (in parentheses) obtained fitting single-trait models were W365 (0.49), W450 (0.52), SC365 (0.68) and SC450 (0.66). Estimates of means, modes and medians for genetic parameters obtained from marginal posterior distributions were similar for all traits. The W365 and SC365 can be considered as suitable traits to be included as selection criteria in genetic improvement programs, not only because of their relatively high heritabilities but also due to the fact that they can be measured when the animals are relatively young compared to the corresponding traits W450 and SC450. The Bayesian approach appears to be an appropriate alternative for estimating genetic parameters, and has the advantage over point estimation methods of allowing inferences on marginal posterior distributions. <![CDATA[<B>A deterministic simulation study of embryo marker-assisted selection for age at first calving in Nellore (<I>Bos indicus</I>) beef cattle</B>]]> We used deterministic simulation of four alternative multiple ovulation and embryo manipulation (MOET) closed nucleus schemes to investigate the benefits of using marker-assisted selection (MAS) of Nellore (Bos indicus) beef cattle embryos prior to transplantation to reduce the age at first calving (AFC). We found that MAS resulted in increased genetic gain as compared to selection without AFC quantitative trait loci (AFC-QTL) information. With single-stage selection the genetic response (GR) increased as follows: GR = 0.68% when the AFC-QTL explained 0.02 of the AFC additive genetic variance (sigma2A); GR = 1.76% for AFC-QTL explaining 0.05 sigma2A; GR = 3.7% for AFC-QTL explaining 0.1 sigma2A; and GR = 55.76% for AFC-QTL explaining 0.95 sigma2A. At the same total selected proportion, two-stage selection resulted in less genetic gain than single stage MAS at two-years of age. A single stage selection responses of > 95% occurred with pre-selected proportions of 0.4 (0.1 sigma2A explained by AFC-QTL), 0.2 (0.3 sigma2A explained by AFC-QTL) and 0.1 (0.5 sigma2A explained by AFC-QTL), indicating that the combined use of MAS and pre-selection can substantially reduce the cost of keeping recipient heifers in MOET breeding schemes. When the number of recipients was kept constant, the benefit of increasing embryo production was greater for the QTL explaining a higher proportion of the additive genetic variance. However this advantage had a diminishing return especially for QTL explaining a small proportion of the additive genetic variance. Thus, marker assisted selection of embryos can be used to achieve increased genetic gain or a similar genetic response at reduced expense by decreasing the number of recipient cows and number of offspring raised to two-years of age. <![CDATA[<B>Polymorphism of exon 2-3 of bovine major histocompatibility complex class I BoLa-A gene</B>]]> The exon 2-3 region of bovine major histocompatibility complex (MHC) class I BoLa-A gene was investigated for polymorphisms in three breeds of cattle originated in the Indian subcontinent namely Sahiwal, Tharparkar, Hariana, as well as crossbred (Bos taurus x Bos indicus) cattle and Jersey, the exotic breed (Bos taurus). The PCR amplified fragment of 714 bp showed distinct DdeI-, TaqI- and HinfI- RFLP patterns, thus confirming a higher degree of polymorphism in this region. To our knowledge this is the first report of HinfI restriction patterns for BoLa-A exon 2-3. The sequencing results revealed a number of nucleotide substitutions in this region, which resulted in amino acid changes. The present investigation confirmed that MHC class I BoLa-A exon 2-3 is highly polymorphic in cattle. <![CDATA[<B>Natural triploidy in <I>Leporinus</I> cf. <I>elongatus</I> bearing sex chromosomes</B>]]> Although several cases of natural triploidy in fish have already been described, spontaneous polyploidy in species with differentiated sex chromosomes are rare. We report the occurrence of a triploid fish (3n = 81) Leporinus cf. elongatus, a species characterized by a highly differentiated ZZ/ZW sex chromosome system, from the São Francisco river. The occurrence of a ZZZ triploid adult indicates the viability of this chromosome constitution in this fish. <![CDATA[<B>Microsatellite variability analysis in farmed catfish (<I>Ictalurus punctatus</I>) from Tamaulipas, Mexico</B>]]> Analysis of cultured catfish from six farms in Tamaulipas, Mexico was achieved using a combination of microsatellite PCR analysis and semiautomatic fluoresce-based detection, in order to provide a first assessment of the genetic variability on cultured catfish in Mexico. Five microsatellites showed extensive polymorphism with allele numbers ranging from 10 and 20. Overall observed heterozygosity at each locus ranged between 0.76 and 0.91 and the average polymorphic information content (PIC) for the five loci was 0.811, indicating that these loci can be used for studies of paternity identification, linkage and population genetics. On the basis of the F ST values (F ST = 0.03829; p = 0.00000) it appears that there was a small amount of genetic differentiation between the channel catfish stocks. The high intrapopulation allelic diversity was the most remarkable parameter. <![CDATA[<B>Quantification of bovine cytokine gene expression using real-time RT-PCR methodology</B>]]> T cells produce cytokines that affect host response to infection. This paper reports real-time RT-PCR conditions and validation steps for accurate quantification of Bos indicus cytokines, interleukin (IL)-2, IL-4, IL-8, IL12p-35, IL-13, tumoral necrosis factor (TNF)-alpha, interferon (IFN)-gamma, monocyte chemoattractant proteins (MCP)-1 and MCP-2, and the glycoprotein mucin (MUC)-1 in two groups of Nelore cattle, one resistant and the other susceptible to gastrointestinal nematode infections. RPL-19 was shown to be an ideal internal control gene, since its expression was constant across treatments and presented lower variation when compared to the GAPDH gene. The optimized conditions established in the present study can be used to determine the immune response of cattle under different experimental conditions, such as viral, bacterial and parasite infections. <![CDATA[<B>Factor XI deficiency in Indian <I>Bos taurus</I>, <I>Bos indicus</I>, <I>Bos taurus x Bos indicus</I> crossbreds and <I>Bubalus bubalis</B></I>]]> We investigated the occurrence of Factor XI (FXI) deficiency syndrome in the following Indian dairy animals: Bos taurus Holstein-Friesian and Jersey cattle, Bos indicus Indian cattle breeds, B. taurus x B. indicus crossbreds and the river buffalo Bubalus bubalis. Factor XI deficiency is an autosomal recessive bleeding disorder known to affect Holstein cattle worldwide. A total of 1001 dairy animals, mainly bulls, were genotyped to detect the mutation within exon 12 of the gene encoding for factor XI. Two Holstein bulls were detected as heterozygous (carrier) for FXI deficiency, giving a carrier frequency of 0.6% in Indian Holstein cattle. None of the other cattle or buffalo breeds was found to be a carrier for FXI. Sequence comparison between normal and heterozygous animals revealed that there is a 77 base pair insertion fragment (AT (A)29 TAAAG (A)27 GAATTATTAATTCT) within exon 12 of the FXI gene. Both sequences were submitted to the National Center for Biotechnology Information (NCBI) GenBank and assigned the accession numbers DQ438908 for normal Holstein Friesian animals and DQ438909 for heterozygous Holstein Friesian animals. <![CDATA[<B>Phenotypic recurrent selection in the common bean (<I>Phaseolus vulgaris</I> L) with carioca-type grains for resistance to the fungi <I>Phaeoisariopsis griseola</B></I>]]> The efficiency of recurrent selection was assessed in obtaining common bean (Phaseolus vulgaris L) plant lines resistant to the phytopathogenic fungi Phaeoisariopsis griseola. The base bean population was obtained from the partial diallel between seven lines with carioca-type grains and 10 sources of resistance to P. griseola. The plants most resistant to the pathogen were selected in the F2 (S0) generation of the populations (C-0). The best S0:1 plants that presented carioca-type grains were intercrossed to obtain cycle I (C-I). The same procedure was adopted to obtain cycles C-II to C-V. In each recurrent selection cycle, S0:1 progenies selected were also assessed in experiments carried out in Lavras, Brazil, always using as check the Carioca MG (susceptible to P. griseola) and Pérola (tolerant) cultivars. The response to selection for resistance to the pathogen was estimated from the general mean of the S0:1 progenies from each selective cycle compared to the susceptible check Carioca MG. The estimated gain was 6.4% per cycle and the indirect response in grain yield by selection for resistance to the pathogen was 8.9% per cycle. The variability detected among the progenies in the last selective cycles enabled the prediction of additional responses to recurrent selection. <![CDATA[<B>Allelic relationships of anthracnose (<I>Colletotrichum lindemuthianum</I>) resistance in the common bean (<I>Phaseolus vulgaris</I> L.) cultivar Michelite and the proposal of a new anthracnose resistance gene, <I>Co-11</B></I>]]> The genetic resistance of Phaseolus vulgaris L. cultivar Michelite to races 8 and 64 of Colletotrichum lindemuthianum, causal agent of bean anthracnose, was characterized. Crosses were made between Michelite and Mexico 222 cultivars and the F2 population was inoculated with race 64 in order to study the inheritance of resistance to anthracnose in Michelite. The segregation of F2 population fitted in a ratio of 3R:1S, showing the presence of a dominant gene in Michelite gene conditioning resistance to race 64. Allelism tests were conducted with F2 populations derived from crosses between Michelite and AB 136, AND 277, BAT 93, Cornell 49-242, G 2333, Kaboon, Mexico 222, Michigan Dark Red Kidney (MRDK), Ouro Negro, Perry Marrow, PI 207262, TO, TU, and Widusa. All the cultivars (except Mexico 222) were resistant to race 64. While F2 derived from the Michelite x Mexico 222 was inoculated with race 8. Additionally, allelism tests indicated that the gene present in Michelite is independent from Co-1, Co-2, Co-3, Co-4, Co-5, Co-6, Co-7, Co-9 and Co-10 genes. The monogenic inheritance observed in Michelite and the independence of this gene from those previously characterized allow the authors to propose that the anthracnose resistant gene in Michelite should be named Co-11. <![CDATA[<B>Genetic control of orange hilum corona of carioca beans (<I>Phaseolus vulgaris</I>)</B>]]> The purpose of this research was to elucidate the genetic control of orange corona color in carioca common beans (Phaseolus vulgaris). We made four crosses between carioca group cultivars that differed in respect to the presence or absence of an orange hilum corona color. The F2, F3, F1BC11, F1BC21, F2BC11 and F2BC21 phenotypic segregations were evaluated with a chi-square test which fitted with the hypothesis that one gene with a dominant allele is responsible for the orange corona color. All generations resulting from the four different crosses showed segregation patterns which agreed with the expected proportions. Our results show that the dominant G allele controls orange corona color in the carioca bean group. <![CDATA[<B>Distribution of a <I>Ty</I>3/<I>gypsy</I>-like retroelement on the A and B-chromosomes of <I>Cestrum strigilatum</I> Ruiz & Pav. and <I>Cestrum intermedium</I> Sendtn. (Solanaceae)</B>]]> Retroelements are a diversified fraction of eukaryotic genomes, with the Ty1/copia and Ty3/gypsy groups being very common in a large number of plant genomes. We isolated an internal segment of the Ty3/gypsy retroelement of Cestrum strigilatum (Solanaceae) using PCR amplification with degenerate primers for a conserved region of reverse transcriptase. The isolated segment (pCs12) was sequenced and showed similarity with Ty3/gypsy retroelements of monocotyledons and dicotyledons. This segment was used as probe in chromosomes of C. strigilatum and Cestrum intermedium. Diffuse hybridization signals were observed along the chromosomes and more accentuated terminal signals in some chromosome pairs, always associated with nucleolus organizer regions (NORs). The physical relationship between the hybridization sites of pCs12 and pTa71 ribosomal probes was assessed after sequential fluorescence in situ hybridization (FISH). Hybridization signals were also detected in the B chromosomes of these species, indicating an entail among the chromosomes of A complement and B-chromosomes. <![CDATA[<B>Partial diallel analysis of agronomic characters in rice (<I>Oryza sativa</I> L.)</B>]]> Rice (Oryza sativa L.) breeding seeking to combine high productivity and cold tolerance for the temperate Latin America region is an important challenge. We estimated some useful parameters which can be used to investigate the genetic control of agronomic characters in crosses combining cold tolerance and productivity. A partial diallel design was used in crosses between six tropical indica rice cold susceptible genotypes (group 1) and seven japonica or indica/japonica cold tolerant rice genotypes (group 2). Parents and crosses were evaluated for agronomic characters under field conditions in two different experiments in 2005. The results showed significant mid-parent heterosis for all characters (plant height, tiller number, days to 50% flowering, panicle length, grains per panicle, sterility, and one-hundred grain weight). The predominant direction of dominance effects was negative for days to 50% flowering, and positive for all the other characters. General combining ability (GCA) and specific combining ability (SCA) were significant for all characters, although the GCA effects of the two groups were more important than the SCA effects. <![CDATA[<B>Comparative linkage mapping of <I>Oryza glumaepatula</I> and <I>Oryza sativa</I> interspecific crosses based on microsatellite and expressed sequence tag markers</B>]]> Molecular linkage maps representing the rice genome have been an important tool for breeding programs because they allow the elucidation of polygenic traits and are an efficient tool for monitoring wild introgressions in interspecific crosses. Common markers among rice genetic maps are important in defining the homology of chromosomes and the synteny between genomic target regions. We used 148 markers (expressed sequence tags, microsatellites and single nucleotide polymorphisms) to construct a molecular linkage map based on co-dominant markers for an interspecific backcross population using a wild rice (Oryza glumaepatula) from Brazil and performed a comparative analysis with other interspecific maps. The comparative analysis revealed a Spearman correlation index of 0.86 for marker order conservation to a previous map constructed for an interspecific cross using the same wild parent. Approximately 90% of markers common to other interspecific maps kept the same order. These results indicate that it will be possible to generate a unique genetic map using the wild donor and that it may be a helpful tool for breeding programs because plants derived from different interspecific populations can be rapidly scanned using markers associated with useful wild traits. <![CDATA[<B>Development of two sequence-specific PCR markers linked to the <I>le</I> gene that reduces pod shattering in narrow-leafed Lupin (<I>Lupinus angustifolius L.</I>)</B>]]> Wild types of narrow-leaf lupin (Lupinus angustifolius L.) have seed pods that shatter upon maturity, leading to the loss of their seeds before or during the harvest process. Two recessive genes have been incorporated into domesticated cultivars of this species to maximize harvest-ability of the produce. One of these genes is called lentus (le). Two microsatellite - anchored fragment length polymorphism (MFLP) candidate markers were identified as closely linked to the le gene in a recombinant inbred line (RIL) population derived from a domesticated x wild type cross. The candidate MFLP markers were isolated from the gel, re-amplified by PCR, cloned and sequenced. The MFLP polymorphisms were converted into sequence-specific PCR-based markers. Linkage analysis by MapManager indicated that one of the markers, LeM1, was 2.6 centiMorgans (cM) and the other, LeM2, was 1.3 cM from the gene, with both being on the same side. The correlation between the marker genotype and the plant phenotype for the le gene is 95% for the Australian cultivars, and approximately 36% on wild types tested. These markers may be useful in marker assisted selection for the le gene when introgressing wild material into lupin breeding programs. <![CDATA[<B>Chromosomal locations of the maize (<I>Zea mays</I> L.) <I>HtP</I> and <I>rt</I> genes that confer resistance to <I>Exserohilum turcicum</B></I>]]> We used 125 microsatellite markers to genotype the maize (Zea mays L.) near isogenic lines (NIL) L30HtPHtPRtRt and L30htphtpRtRt and the L40htphtprtrt line which contrast regarding the presence of the recently described dominant HtP and the recessive rt genes that confer resistance to Exserohilum turcicum. Five microsatellite markers revealed polymorphisms between the NIL and were considered candidate linked markers for the HtP resistance gene. Linkage was confirmed by bulked segregant sample (BSS) analysis of 32 susceptible and 34 resistant plants from a BC1F1 population derived from the cross (L30HtPHtPRtRt x L40htphtprtrt) x L40htphtprtrt. The bnlg198 and dupssr25 markers, both located on maize chromosome 2L (bin 2.08), were polymorphic between bulks. Linkage distances were estimated based on co-segregation data of the 32 susceptible plants and indicated distances of 28.7 centimorgans (cM) between HtP and bnlg198 and 23.5 cM between HtP and dupssr25. The same set of susceptible plants was also genotyped with markers polymorphic between L30HtPHtPRtRt and L40htphtprtrt in order to find markers linked to the rt gene. Marker bnlg197, from chromosome 3L (bin 3.06), was found linked to rt at a distance of 9.7 cM. This is the first report on the chromosomal locations of these newly described genes. <![CDATA[<B>Quantitative trait loci mapping of pubescence density and flowering time of insect-resistant soybean (<I>Glycine max</I> L. Merr.)</B>]]> Analysis of antibiosis resistance to common cutworm (Spodoptera litura Fabricius) in soybean (Glycine max (L.) Merr.) has progressed significantly, but the immediate cause remains unknown. We performed quantitative trait loci (QTL) analysis of pubescence density and plant development stage because these factors are assumed to be the immediate cause of resistance to cutworm. The QTLs for pubescence appeared to be identical to the previously detected the Pd1 and Ps loci controlling pubescence density. We found no candidate loci for flowering time QTLs, although one could be identical to the gene governing the long-juvenile trait or to the E6 loci controlling maturity. None of the QTLs overlapped with the QTLs for antibiosis resistance. <![CDATA[<B><I>Astylus variegatus</I> (Coleoptera, Melyridae)</B>: <B>Cytogenetic study of a population exposed to agrochemical products</B>]]> In this work we describe the cytogenetic analyses performed in specimens of Astylus variegatus (Germar, 1824) collected in two localities: one area of natural vegetation and one of agricultural crops, where agrochemical products were used. Astylus variegatus had karyotypes with 2n(male) = 16+Xy p and 2n (female) = 16+XXp, with exclusively metacentric chromosomes. Pachytene spermatocytes showed synapsed autosomal bivalents and non-associated sex chromosomes. In diplotene, the autosomal bivalents exhibited one or two terminal chiasmata and the Xy p had a typical parachute configuration. In meiotic cells of some specimens, an extra chromosome, interpreted as a B chromosome, was observed. C-banding showed constitutive heterochromatin in the pericentromeric region of all chromosomes, with the exception of the y p. Silver nitrate staining revealed one nucleolus organizer region (NOR) on the terminal region of the short arm of the second autosome pair. Silver staining of meiotic cells confirmed the NOR pattern detected in mitotic cells and revealed an argentophilous material on the Xy p. A cytogenetic comparison between the two populations of A. variegatus showed a statistically significant divergence (chi2 = 117.10; df = 1) in the number of aneuploid cells and a higher frequency of B chromosome in the population exposed to agrochemicals. <![CDATA[<B>CUG-BP and 3'UTR sequences influence PARN-mediated deadenylation in mammalian cell extracts</B>]]> Several mRNAs have been shown to exhibit distinct patterns of poly(A) shortening prior to their decay in vivo. In this study, we show that individual transcripts also demonstrate distinct patterns of deadenylation in in vitro systems derived from HeLa and Jurkat T cell cytoplasmic extracts. The major patterns observed were slow/synchronous and fast/asynchronous poly(A) tail shortening. For all RNA substrates tested, PARN was shown to be the enzyme responsible for the deadenylation patterns that were observed. Sequences in the 3' untranslated regions influenced the deadenylation pattern. Using a fragment of the 3'UTR of the c-fos mRNA as a model, the interaction of CUG-BP, the human homolog of EDEN-BP - a protein previously implicated in regulated deadenylation in Xenopus oocytes - was shown to be associated with changes in PARN-mediated deadenylation patterns. Our results suggest that association of CUG-BP with 3'UTR sequences can modulate the activity of the PARN deadenylase in mammalian cell extracts. <![CDATA[<B>Clastogenicity of <I>Piper cubeba</I> (Piperaceae) seed extract in an <I>in vivo</I> mammalian cell system</B>]]> The plant Piper cubeba is widely distributed in tropical and subtropical regions and is used medically for various purposes but has not yet been evaluated for genotoxicity. We used male and female Swiss mice and Wistar rats and the comet assay and micronucleus test to investigate the mutagenic potential of a crude extract of P. cubeba seeds. The rodents were administered 0.5 g kg-1, 1.0 g kg-1 and 1.5 g kg-1 of the extract by gavage. For the Swiss mice, peripheral blood was collected 24 h after treatment for the comet assay, and at 48 and 72 h for the micronucleus test. For the Wistar rats, peripheral blood and hepatic cells were collected for the comet assay and bone marrow cells were collected for the micronucleus test 24 h after treatment. At 1.5 g kg-1, the highest dose tested, the extract induced a statistically significant increase in both the mean number of micronucleated polychromatic erythrocytes and the level of DNA damage in the rodent cell types analyzed. Under our experimental conditions, the P. cubeba seed extract was genotoxic in vivo when administered orally to mice and rats. <![CDATA[<B>On extending the Hardy-Weinberg law</B>]]> This paper gives a general mating system for an autosomal locus with two alleles. The population reproduces in discrete and non-overlapping generations. The parental population, the same in both sexes, is arbitrary as is that of the offspring and the gene frequencies of the parents are maintained in the offspring. The system encompasses a number of special cases including the random mating model of Weinberg and Hardy. Thus it demonstrates, in the most general way possible, how genetic variation can be conserved in an indefinitely large population without invoking random mating or balancing selection. An important feature is that it provides a mating system which identifies when mating does and does not produce Hardy-Weinberg proportions among offspring. <![CDATA[<B>Genetic divergence between populations of the stingless bee uruçu amarela (<I>Melipona rufiventris</I> group, Hymenoptera, Meliponini)</B>: <B>is there a new <I>Melipona</I> species in the Brazilian state of Minas Gerais?</B>]]> Allozyme, microsatellite and random amplified polymorphic DNA (RAPD) molecular markers were used to investigate the within and between population genetic variability and between population genetic differentiation of the Brazilian stingless bee uruçu amarela (nominally Melipona rufiventris Lepeletier, 1836) present in savanna and Atlantic forest habitats of the Brazilian state of Minas Gerais (MG). We found low levels of within population variability, although there were a large number of private alleles that specifically characterized these populations. The F ST values indicated a high level of genetic diversity between populations. Analysis of molecular variance (AMOVA) showed a high degree of population differentiation between the savanna and Atlantic forest habitats, confirmed by population pairwise F ST data. Principal coordinates analysis and unweighted pair-group method using an arithmetic average (UPGMA) dendrograms also confirmed that in Minas Gerais the savanna populations (M. rufiventris) were genetically distinct from those present in the Atlantic forest (M. mondury). In addition, populations from locations near the towns of Dom Bosco and Brasilândia de Minas were genetically different from those collected in other localities in the savanna. Our data indicate that populations of uruçu amarela found in the savanna and Atlantic forest habitats of Minas Gerais state should be treated separately for conservation purposes and that special attention should be given to the populations found in the region of Dom Bosco and Brasilândia de Minas until their taxonomic status is clarified. <![CDATA[<B>Distribution of the <I>Bari-I</I> transposable element in stable hybrid strains between <I>Drosophila melanogaster</I> and <I>Drosophila simulans</I> and in Brazilian populations of these species</B>]]> We analyzed the distribution of the Bari-I transposable element in Drosophila melanogaster (IN(1)AB), its sibling species Drosophila simulans (C167.4) and in eight hybrid strains derived from initial crosses involving D. simulans females and D. melanogaster males of the above cited strains as well as in Brazilian populations of these species. Polymerase chain reaction (PCR) data showed the presence of the Bari-I element among species populations and hybrid strains. Hybridization with a 703 bp probe homologous to the Bari-I sequence showed that the number of Bari-I copies in D. melanogaster IN(1)AB was higher than in D. simulans C167.4 strains. Hybrid strains presented Bari-I sequences related to both parental species. In addition some strains displayed a Bari-I sequence that came from D. melanogaster, suggesting introgression of D. melanogaster genetic material in the background of D. simulans. In contrast, some hybrids showed deletions of D. simulans Bari-I sequences. <![CDATA[<B>Challenges and prospects of population genetic studies in terns (Charadriiformes, Aves)</B>]]> Little information is available about the population structure of communally nesting terns (Sternidae) and skimmers (Rynchopidae) throughout the world. In order to fill this gap, a survey of molecular markers was carried out for six species of terns (Anous stolidus, Sterna hirundinacea, S. fuscata, S. superciliaris, Thalasseus maximus and Phaetusa simplex) and one species of skimmer (Rynchops niger). First, we describe the results of the construction of genomic DNA libraries and document problems encountered during this procedure. Secondly, we tested the cross-amplification of 18 microsatellite loci previously described for related species (the number of polymorphic loci ranged from three to seven). Thirdly, we tested the usefulness of mtDNA (control region, ND2, Cytochrome b and ATPase 6/8) for phylogeographic studies in this group of birds. The occurrence of nuclear copies of the mitochondrial control region is reported. Nucleotide divergence in the mtDNA genes analyzed ranged from 0.0 to 0.006. Despite the difficulties associated with the selection of variable markers in this group of seabirds, we were able to select polymorphic markers for each species tested and we anticipate these results will help the development of genetic studies concerning important biological questions in terns. <![CDATA[<B>Reproductive strategies and genetic variability in tropical freshwater fish</B>]]> We estimated the genetic variability of nine fish species from the Brazilian upper Paraná River floodplain (Astyanax altiparanae, Hoplias malabaricus, Leporinus lacustris, Loricariichthys platymetopon, Parauchenipterus galeatus, Pimelodus maculatus, Rhaphiodon vulpinus, Roeboides paranensis and Serrasalmus marginatus) based on data for 36 putative allozyme loci obtained using corn starch gel electrophoresis of 13 enzymatic systems: aspartate aminotransferase (EC, acid phosphatase (EC, esterase (EC, glycerol-3-phosphate dehydrogenase (EC, glucose-6-phosphate dehydrogenase (EC, glucose-6-phosphate isomerase (EC, Iditol dehydrogenase (EC, isocitrate dehydrogenase - NADP+ (EC, L-lactate dehydrogenase (EC, malate dehydrogenase (EC, malate dehydrogenase-NADP+ (EC, phosphoglucomutase (EC and superoxide dismutase, (EC The mean expected heterozygosity varied from zero to 0.147. When data from the literature for 75 species of tropical fish were added to the nine species of this study, the heterozygosity values differed significantly among the groups of different reproductive strategies. The highest mean heterozygosity was for the non-migratory without parental care, followed by the long-distance migratory, and the lowest mean was for the non-migratory with parental care or internal fecundation. <![CDATA[<B>The use of PCR-RFLP as an identification tool for three closely related species of rodents of the genus <I>Akodon</I> (Sigmodontinae, Akodontini)</B>]]> Three cryptic species of the rodent Akodon (A. cursor, A. montensis and Akodon sp.) were analyzed. The two former species are sympatric in the Brazilian states of São Paulo, Rio de Janeiro and Minas Gerais, where hybrids have already been found. The third species, Akodon sp., occurs in an isolated area in Central Brazil. The identification of these species is difficult by the need of living animals. At present, karyotyping is the only method used in the identification of specimens. We used PCR-RFLP of the mitochondrial cytochrome gene to test the distinctiveness of the three species, which was confirmed by the absence of shared species-specific haplotypes. We also detected a geographical pattern of haplotypes distribution with highly polymorphic populations of A. cursor from Espírito Santo and of A. montensis from Rio Grande do Sul. <![CDATA[<B>Expression of D-type cyclins in differentiating cells of the mouse spinal cord</B>]]> The D-type cyclins form complexes with the cyclin dependent (CD) kinases CDK4 and CDK6 and promote the G1-S phase transition of the cell cycle by antagonizing the retinoblastoma suppresser protein pRB. In the developing nervous system D-type cyclins show spatially and temporally dynamic patterns of expression. We demonstrated that cyclin D1 was transiently expressed in differentiating spinal cord ventral interneurons while cyclin D3 protein was expressed in differentiating motor neurons and dorsal interneurons. This expression of cyclin D3 in neurons of the mantle zone was extended to all regions of the spinal cord at E15.5. The results suggest that cyclin D1 and D3 have specific functions in differentiating neurons. Similarly, in the developing midbrain-hindbrain region the D-type cyclins were expressed in different subsets of cells. Our results argue in favor of different functions for D-type cyclins during proliferation and differentiation of neural progenitors. <![CDATA[<B>The systemic concept of the gene, at age fifteen, and comments on C.N. El-Hani's article 'Between the cross and the sword</B>: <B>the crisis of the gene concept'</B>]]> The D-type cyclins form complexes with the cyclin dependent (CD) kinases CDK4 and CDK6 and promote the G1-S phase transition of the cell cycle by antagonizing the retinoblastoma suppresser protein pRB. In the developing nervous system D-type cyclins show spatially and temporally dynamic patterns of expression. We demonstrated that cyclin D1 was transiently expressed in differentiating spinal cord ventral interneurons while cyclin D3 protein was expressed in differentiating motor neurons and dorsal interneurons. This expression of cyclin D3 in neurons of the mantle zone was extended to all regions of the spinal cord at E15.5. The results suggest that cyclin D1 and D3 have specific functions in differentiating neurons. Similarly, in the developing midbrain-hindbrain region the D-type cyclins were expressed in different subsets of cells. Our results argue in favor of different functions for D-type cyclins during proliferation and differentiation of neural progenitors. <![CDATA[<B>Comments on R.C. Guimarães' 'The systemic concept of the gene at age fifteen'</B>]]> The D-type cyclins form complexes with the cyclin dependent (CD) kinases CDK4 and CDK6 and promote the G1-S phase transition of the cell cycle by antagonizing the retinoblastoma suppresser protein pRB. In the developing nervous system D-type cyclins show spatially and temporally dynamic patterns of expression. We demonstrated that cyclin D1 was transiently expressed in differentiating spinal cord ventral interneurons while cyclin D3 protein was expressed in differentiating motor neurons and dorsal interneurons. This expression of cyclin D3 in neurons of the mantle zone was extended to all regions of the spinal cord at E15.5. The results suggest that cyclin D1 and D3 have specific functions in differentiating neurons. Similarly, in the developing midbrain-hindbrain region the D-type cyclins were expressed in different subsets of cells. Our results argue in favor of different functions for D-type cyclins during proliferation and differentiation of neural progenitors.