ABSTRACT. Wheat is notorious for callus induction, which is a major hindrance in direct gene transfer and consequently for genetic improvement programs. In order to provide a successful platform for gene transformation, good callus quantity and quality is impor­tant. We investigated the variation in callus induction capabilities of Pakistani wheat cultivars and measured the reducing sugar con­tent in the induced calluses. Ten elite wheat varieties, developed and cultivated in Pakistan were selected on the basis of agronomic and stress tolerance parameters. Significant differences were found between and among wheat cultivars for callus induction response, shoot length and callus quality. The callus induction responses of Punjab-81, Punjab-96 and Zarghoon-79 were found to be the best among the 10 varieties. The induced calluses were of two types, em­bryogenic (hard) and non-embryogenic (soft). The seeds gave good germination. The highest reducing sugar concentration was found in cultivar Sutlaj-86, which needs to be tested for stress resistance, a measure of its utility for genetic engineering programs. The relative callus induction rate and reducing sugar content of the wheat culti­vars were found to be genotype-dependent.

ABSTRACT. Development of hereditary hemochromatosis is asso­ciated with the C282Y, H63D or S65C mutations in the hemochro­matosis gene. Though there is extensive knowledge about the former two, there is little information on the mechanism of action and the allelic frequency of the S65C mutation. We examined the prevalence of the S65C mutation of the hemochromatosis gene in Brazilians with clinical suspicion of hereditary hemochromatosis. Genotyping for this mutation was carried out in 633 individuals with clinical suspicion of hereditary hemochromatosis, using the polymerase chain reaction, followed by enzymatic digestion. The sample comprised 77.1% men and 22.9% women, giving a ratio of approximately 3:1; the mean age was 48.8 + 13.8 years. More than half (57.3%) of the individuals in the sample were 41 to 60 years old. The frequency of heterozygotes for this mutation was 0.016; no homozygous mutant patients were found. This is the first analysis of the S65C mutation in individuals suspected of having hereditary hemochromatosis in Brazil.

ABSTRACT. In order to increase the efficiency of cacao tree resistance to witches’ broom disease, which is caused by Moniliophthora perniciosa (Tricholomataceae), we looked for molecular markers that could help in the selection of resistant cacao genotypes. Among the different markers useful for developing marker-assisted selection, single nucleotide polymorphisms (SNPs) constitute the most common type of sequence difference between alleles and can be easily detected by in silico analysis from expressed sequence tag libraries. We report the first detection and analysis of SNPs from cacao-M. perniciosa interaction expressed sequence tags, using bioinformatics. Selection based on analysis of these SNPs should be useful for developing cacao varieties resistant to this devastating disease.

ABSTRACT. We investigated the genetic diversity of 63 wheat genotypes, composed of 48 accessions and 15 varieties, using 56 polymorphic simple sequence repeat primers. One hundred and eighty-six loci were found, with a mean of 131.26 alleles per locus. Cluster analysis based on microsatellite allelic diversity discrimi­nated the accessions and varieties into different clusters; genetic di­versity was the highest between variety Kohistan-97 and accession number 011512, giving a genetic similarity value of 0.4198. Acces­sion numbers 011484 and 011356 gave a genetic similarity value of 0.9589, indicating that these accessions were 95.89% similar. We found that microsatellite markers could characterize and discrimi­nate all of the genotypes; more primers could be used for saturation of different regions in further studies.

ABSTRACT. The myostatin gene, also known as GDF8 (growth dif­ferentiation factor 8), is located on bovine chromosome 2 (BTA2); it has three exons and two introns. Myostatin is specifically expressed during embryonic development and in adult skeletal muscle, func­tioning as a negative regulatory protein. Several cattle breeds (Pied­montese, Belgian Blue and Blond’Aquitaine, and others) show poly­morphisms in this gene; these polymorphisms are directly related to the double muscling phenotype. We looked for polymorphisms in the Nellore cattle myostatin gene and compared them with those known for taurine breeds. Seven regions, covering the three exons of this gene, were amplified by polymerase chain reaction and se­quenced, including the untranslated region. DNA from 30 adult Nel­lore animals was collected; DNA sequencing revealed three, seven and four polymorphisms in exons 1, 2 and 3, respectively. We found previously reported polymorphisms, as well as several new ones; for instance, 37 polymorphisms were found in the untranslated re­gion segment, and in introns 1 and 2 there were one and three poly­morphisms, respectively. The high degree of allelic heterogeneity in the myostatin gene could be related to its high mutation rate; it also could be the result of a long history of artificial selection for meat production, which has probably favored such modifications and maintained them in cattle populations. These polymorphisms identified in Nellore cattle could be useful for breeding programs.

ABSTRACT. Scaptotrigona xanthotricha has a wide geographic distribution in the Brazilian Atlantic rainforest. One population from southeast and two from northeast Brazil were analyzed and were found to have chromosome polymorphisms. Although the chromo­some number 2n = 34 is conserved in this species, karyotypic analy­sis revealed clear differences between the three populations. Congru­ent and ubiquitous multiple nucleolus organizer regions, heterochro­matin and CMA₃-positive blocks were found. The variations suggest that this species is in a process of genetic differentiation. This dif­ferentiation process might have been enhanced by restricted nesting preferences, combined with recent extensive fragmentation of the Atlantic rainforest, which limits gene flow between populations.

ABSTRACT. Previously, we isolated two strains of spontaneous oxidative (SpOx2 and SpOx3) stress mutants of Lactococcus lactis subsp cremoris. Herein, we compared these mutants to a parental wild-type strain (J60011) and a commercial starter in experimental fermented milk production. Total solid contents of milk and fermen­tation temperature both affected the acidification profile of the spon­taneous oxidative stress-resistant L. lactis mutants during fermented milk production. Fermentation times to pH 4.7 ranged from 6.40 h (J60011) to 9.36 h (SpOx2); V_max values were inversely proportional to fermentation time. Bacterial counts increased to above 8.50 log10 cfu/mL. The counts of viable SpOx3 mutants were higher than those of the parental wild strain in all treatments. All fermented milk prod­ucts showed post-fermentation acidification after 24 h of storage at 4ºC; they remained stable after one week of storage.

ABSTRACT. At present, studies on microRNA mainly focus on the identification of microRNA genes and their mRNA targets. Although researchers have identified many microRNA genes, relatively few microRNA targets have been identified by experi­mental methods. Computational programs designed for predicting potential microRNA targets provide numerous targets for exper­imental validation. We used a Markov model to examine base-pairing binding patterns of known microRNA targets. Using this model, potential microRNA targets in human species predicted by four well-known computational programs were assessed. Each potential target was assigned a score reflecting consistency with known target binding patterns. Targets with scores higher than the cutoff value would be identified by our model. The predicted targets identified by our model have base-pairing binding pat­terns consistent with known targets. This model was efficient for evaluating the extent to which a potential target was accurately predicted.

ABSTRACT. Wolbachia are bacteria that live inside the cells of a large number of invertebrate hosts and are transmitted from infected females to their offspring. Their presence is associated with cytoplasmic incompatibility in several species of Drosophila. Cytoplasmic incompatibility results when the sperm of infected males fertilize eggs of uninfected females, causing more or less intense embryonic mortality (unidirectional incompatibility). This phenomenon also appears in crosses between populations infected with different Wolbachia strains (bidirectional incompatibility). The influence of Wolbachia infection on host populations has attracted attention as a potentially rapid mechanism for development of reproductive isolation and subsequent speciation. We examined the influence of this bacterium on reproductive isolation in interspecific crosses between Drosophila melanogaster and D. simulans. We found that Wolbachia infection negatively affected these two species in homospecific crosses. However, in interspecific crosses, it only influenced sexual isolation, as infected females more frequently hybridized than females free of infection; postzygotic reduction of fitness (bidirectional cytoplasmic incompatibility) was not detected. This would be explained by the existence of several modes of rescue systems in these two species, reducing cytoplasmic incompatibility between them. Wolbachia does not appear to cause reproductive isolation between these two species.

ABSTRACT. We investigated seven distance measures and 14 simi­larity coefficients in a set of observations of variables of the ‘yellow’ passion fruit plant (Passiflora edulis Sims), submitted to multivari­ate analyses (distance, projection and grouping). Fourteen genotypes were characterized, based on DNA amplification with 16 random am­plified polymorphic DNA primers and the assessment of nine fruit physical-chemical descriptors. The distance measurements and the similarity coefficients were compared by the Spearman correlation test, projection in two-dimensional space and grouping efficiency, using five grouping methods; the genotype ranking varied with the different techniques. Coler-Rodger distance measures, Euclidean distance square measures and Yule similarity coefficients proved to be inadequate for projection in two-dimensional space or for group­ing matrices. Regardless of the origin of the distance matrix, the un­weighted pair group method with arithmetic mean grouping method proved to be the most adequate. The various distance measurements, similarity coefficients and grouping methods gave different values of distortion, cophenetic correlation and stress; they influence the char­acterization of genetic variability and this should be taken into ac­count for studies of yellow passion fruit plants.

ABSTRACT. Green bean genotypes collected from eastern Turkey were characterized using simple sequence repeat (SSR) markers and morphological traits. Among 12 SSR markers, 10 produced successful amplifications and revealed DNA polymorphisms that were subsequently used to assess genetic relatedness of the genotypes. Based on the number of alleles generated and the probability of identity values, the most informative SSR loci were PVGLND5, PVMEIG, PV-ag001, and PV-ag004. Probably, due to the inbreeding nature of beans, the heterozygosity observed within genotypes was low at most of the SSR loci. The UPGMA dendrogram constructed based on the SSR data yielded two major clusters. The overall genetic distance was around 98%, among the genotypes. This information can be used to help select Turkish green bean lines.

ABSTRACT. Brachiaria humidicola, a species adapted to poorly drained and infertile acid soils, is widely used throughout the tropics. Cytological characterization of 54 accessions of B. humidicola for breeding purposes revealed 2n = 36, 42, and 54 chromosomes. One accession (H030), with 2n = 42 chromosomes, showed a different meiotic behavior. In most accessions from the genus Brachiaria previously studied, the basic chromosome number is x = 9, but the putative basic number in H030 appears to be x = 6. Since six univalent chromosomes were found in diakinesis and metaphase I, and these behaved as laggards in anaphase I, it was hypothesized that both genitors were derived from x = 6, and that this accession is a heptaploid 2n = 7x = 42. The parental genomes did not have the same meiotic behavior, particularly during anaphase, when one genome consisting of six univalents remained as laggards and underwent sister-chromatid segregation. At telophase, 18 segregated chromosomes were found at each pole. The laggard genome did not reach the poles at telophase I or II in time to be included in the nucleus and was eliminated as micronuclei.

ABSTRACT. It is a challenge to understand how development emerged as a mechanism to dismantle and dismiss the intromission of foreign parasites in order to consolidate a higher-level multicellular unit of selection where more heritable variations in fitness, required for complex organization, can be procured. Levels in biological hierarchy genes, networks of genes, chromosomes, cells, organisms, etc., possess heritable variations in fitness to varying degrees, and as such, they function as units of selection in the evolutionary process [Lewontin, (1970). The units of selection. Annu. Rev. Ecol. Syst. 1: 1-18]. To proceed from each of these levels to the next constitutes a major transition in evolutionary history. When analyzing the splendid road epitomized by these transitions in units of selection, it is possible to conceive three processes: firstly, the molecular “recognition” of the “convenience” of exchanging the higher energy cost of cooperating cells with more fitness than single-cell selection (after that first recognition the emergence of cooperation among cells is possible); secondly, the establishment of the mechanisms to regulate conflict, and finally, the regulation of cell differentiation and compartmentalization.

ABSTRACT. Infertility is defined as the inability to conceive a child after one year of regular unprotected intercourse; it is a major health problem affecting about 10-15% of all couples. Infertility is due to a male factor in approximately 50% of cases. The human Y chromosome contains genes necessary for gonadal differentiation into a testis and genes for complete spermatogenesis. We examined the frequency and type of both chromosomal abnormalities and Y chromosome microdeletions in 90 patients with severe male factor infertility and 75 fertile control men. Thirty of the infertile patients had nonobstructive azoospermia, 30 had oligozoospermia and 30 had normozoospermia. Five of 30 were azoospermic, four of 30 were oligozoospermic and two of 30 were normozoospermic with Y chromosome microdeletions. The AZFc locus was the most frequently deleted region (64%). Ten cases with azoospermia, four cases with oligozoospermia and four cases with normozoospermia had chromosomal abnormalities. The 75 men with proven fertility were genetically normal. We conclude that various chromosomal abnormalities and deletions of the Y chromosome can cause infertility; therefore, genetic screening is indicated for infertile patients.

ABSTRACT. The radiotracer technetium-99m methoxyisobutyl isonitrile (^99mTc-MIBI) has been widely used for myocardial blood flow imaging. We investigated the genotoxicity of ^99mTc-MIBI in cultured human lymphocytes at the same concentration used in patients. Radioactivity doses were determined in whole blood at 5 min post-injection of 20 mCi ^99mTc-MIBI in patients. Subsequently, whole blood of human volunteers was incubated with 1, 2.3, 4 or 8 μCi ^99mTc-MIBI. After a 30-min incubation, the lymphocytes were stimulated with a mitogen to assay for micronuclei in cytokinesis-blocked binucleated cells. The frequency of micronuclei in samples treated with this radiopharmaceutical up to 2-fold (8 μCi) the concentration of ^99mTc-MIBI normally found in the blood of patients was not more than in control lymphocyte cultures. We concluded that there is no increased induction of micronuclei in lymphocytes incubated with ^99mTc-MIBI at the radioactivity doses used for diagnostic purposes.

ABSTRACT. Human amniotic fluid is of both maternal and fetal origin; it protects the fetus and provides the environment for growth and development of the fetus. We used a proteomics-based approach for targeting and purifying human phosphate binding protein, a member of the DING family of proteins from amniotic fluid, using Blue Sepharose CL-6B, DEAE-Sephacel and gel filtration chroma­tography. The protein had earlier been reported to be serendipitously purified along with PON1 (paraoxonase 1). It was identified using electro-spray-ionization-time-of-flight mass spectrometry and was found to be human phosphate binding protein. Human phosphate binding proteins have been reported to play a role as phosphate scav­engers and may have a protective function against phosphate-related disorders, such as atherosclerosis, diabetes and kidney stones.

ABSTRACT. Isabgol is a medicinal plant known for its high-quality dietary fiber. The genetics and inheritance of economic characters, such as number of panicles, panicle length, seed yield, and swelling factor, were measured by diallel analysis of F1 progenies from seven parents. The additive component of genetic variance was significant for days to flowering, plant height, branches/plant, peduncle length, panicle length, days to maturity, and swelling factor, whereas the dominance component of genetic variance was significant for all the characters except panicles/plant. Additivity was not significant for all the characters, indicating absence of non-allelic interactions (epistasis) in controlling gene expression. Heritability in the narrow sense was very high for panicle length, days to flowering, and plant height, and moderate for branches/plant, panicles/plant, days to maturity, seed yield, husk yield, and swelling factor. However, the degree of genetic improvement was only high for panicle length, seed yield and husk yield. We conclude that hybridizations, isolation of superior genotypes by sib selection and recurrent selection, and exploitation of hybrid vigor in specific parental-cross combinations are good strategies for isabgol crop improvement.

ABSTRACT. Diallel analysis was used to obtain information on com­bining ability, heterosis, estimates of genetic distances by random amplified polymorphic DNA (RAPD) and on their correlations with heterosis, for the popcorn varieties RS 20, UNB2, CMS 43, CMS 42, Zélia, UEM J1, UEM M2, Beija-Flor, and Viçosa, which were crossed to obtain all possible combinations, without reciprocals. The genitors and the 36 F1 hybrids were evaluated in field trials in Maringá during two growing seasons in a randomized complete block design with three replications. Based on the results, strategies for further studies were developed, including the construction of composites by joining varieties with high general combining ability for grain yield (UNB2 and CMS 42) with those with high general combining ability for popping expan­sion (Zélia, RS 20 and UEM M2). Based on the RAPD markers, UEM J1 and Zélia were the most genetically distant and RS 20 and UNB2 were the most similar. The low correlation between heterosis and genet­ic distances may be explained by the random dispersion of the RAPD markers, which were insufficient for the exploitation of the popcorn genome. We concluded that an association between genetic dissimilar­ity and heterosis based only on genetic distance is not expected without considering the effect of dominant loci.

ABSTRACT. Cocona (Solanum sessiliflorum Dunal) is a solanaceous shrub native to the Amazon region that produces an edible fruit. This species has numerous advantages, particularly a high nutritional value and productivity. However, due to irregular germination and rapid loss of seed viability, there are few plantations for production on a large scale. Development of alternative propagation strategies is essential for the production of homogeneous seedlings of genotypes with superior agronomic performance. We developed techniques for in vitro regeneration of the cocona varieties Santa Luzia and Thaís for large-scale production of healthy plantlets. Twenty days after seeding, seedling segments germinated in vitro were used as explant sources. Three successive experiments were performed: one to test the effect of the explant source and combinations of two growth regulators, auxin (indole acetic acid, IAA) and kinetin (KIN), on the morphogenetic response; another to investigate the effect of the combination of growth regulators on the morphogenetic response of hypocotyl segments, and another to evaluate how sucrose concentration affects the development of adventitious shoots. The best shoot induction was obtained using hypocotyl segments and stem apices, while rhizogenesis was greatest in leaves with a petiole. The number of adventitious shoots per explant on hypocotyl segments increased with 10 and 20 mg/L KIN, combined with 0.02 mg/L IAA in the variety Santa Luzia. Sucrose combined with these growth regulator levels increased the average number of calli; these were optimally produced when 45 g/L sucrose and 0.01 mg/L IAA + 20 mg/L KIN were applied. Only sucrose concentration influenced shoot proliferation in the two S. sessiliflorum varieties, with a maximum at 17.5 g/L.

ABSTRACT. We examined leaf CO₂ assimilation and how it varied among species within the orchid genus Cattleya. Measurements of CO₂ assimilation and maximum quantum yield of PS II (Fv/Fm) were made for mature leaves of nine species using a portable system for photosyn­thesis measurement and a portable fluorometer. Leaf area was measured with an area meter, and the specific leaf mass was determined. DNA of nine Cattleya species and two species of Hadrolaelia was extracted us­ing the CTAB protocol. Each sample was amplified and sequenced using primers for the trnL gene. The phylogenetic analyses, using neighbor-joining and maximum parsimony methods, retrieved a group that included Cattleya and Hadrolaelia species, in which the unifoliate species were separated from the bifoliates. The topologies of the two cladograms showed some similarities. However, C. guttata (bifoliate) was placed in the unifoliate clade in the neighbor-joining tree, while C. warneri (uni­foliate) was not placed in this clade in the maximum parsimony tree. Most Cattleya species keep the leaf stomata closed from 6 am to 4 pm. We suggest that C. elongata, C. tigrina and C. tenuis have C₃-crassu­lacean acid metabolism since they open their stomata around 12 am. The Fv/Fm values remained relatively constant during the measurements of CO₂ assimilation. The same was observed for the specific leaf mass val­ues, although great variations were found in the leaf area values. When the species were grouped using molecular data in the neighbor-joining analysis, no relation was observed with CO₂ assimilation.

ABSTRACT. The mite Varroa destructor is an ectoparasite that is con­sidered a major pest for beekeeping with European honey bees. However, Africanized bee colonies are less threatened by this ectoparasite, because infestation levels remain low in these bees. The low reproductive ability of female mites of the Japanese biotype (J), introduced to Brazil early in the 1970s was initially considered the main factor for the lack of viru­lence of this parasite on Africanized bees. In other regions of the world where the Korean (K) biotype of this mite was introduced, there have been serious problems with Varroa due to the high reproductive potential of the mite. However, a significant increase in the reproductive rate of females of Varroa in Brazil has been recently demonstrated; the cause could be a change in the type of Varroa in the bee colonies. We evalu­ated the prevalence of haplotypes J and K in mite samples collected from the State of Santa Catarina and from the island of Fernando de Noronha in the State of Pernambuco. The analysis of the mitochondrial genome (PCR + RFLP) revealed haplotype K in all samples from Santa Catarina and haplotype J in all samples from Fernando de Noronha. The analysis of microsatellites (nuclear genome) in bees from Fernando de Noronha showed only the specific alleles of haplotype J, while in bees from Santa Catarina, these alleles were found in only 2.8% of the samples. The high frequency of individuals with Korean genetic material is probably to the reason for the current high reproductive capacity of the mite V. destructor recorded in Santa Catarina.

ABSTRACT. Aedes albopictus, a mosquito originally from Southeast Asia, is considered to be one of the main vectors of dengue fever, yel­low fever and other arboviruses. We examined the genetic variability and population structure of 68 individuals of Ae. albopictus collected from five neighborhoods of the city of Manaus, based on the mitochon­drial gene coding for NADH dehydrogenase subunit 5 (ND5). Two hap­lotypes were found, separated by a single mutational event (T ↔ C), with extremely low levels of genetic variability (h = 0.187 ± 0.059; π = 0.00044 ± 0.00014). Based on AMOVA, we concluded that most of the variation (99.08%) occurred within populations, though the levels of variation were not significant. Neutrality tests (Tajima’s D and Fu’s Fs) were non-significant, indicating that these populations are in genetic equilibrium. The most frequent haplotype (H1) is restricted to Brazilian populations of Ae. albopictus, while the rarer haplotype (H2) is shared with populations from the United States and Asia. We suggest that the reduced variability and low genetic structure identified in our study is a consequence of the recent introduction of this species in Manaus, pos­sibly through a founder effect, followed by expansion throughout the city neighborhoods. Genetic similarity would therefore be due to insuf­ficient time to have accumulated genetic differences between the popu­lations of Ae. albopictus and not to extensive gene flow among them.

ABSTRACT. We examined the polymorphisms in the PIT1 gene of 103 pigs and compared their frequencies in the maternal and paternal lineages of the Pietrain and Large White breeds, which have undergone divergent selection for over 30 years. DNA samples extracted from the blood of these animals were amplified by PCR and genotyped by RFLP, using the restriction enzyme RsaI. The data were analyzed with the chi-square test. We found that 57.3% of the animals were genotype AB, among which 26.2% were of the Large White paternal lineage, 18.5% the Pietrain paternal lineage and 12.6% the Pietrain maternal lineage. The AA genotype appeared in 20.4%, of which 7.8% were Large White, 4.8% the Pietrain paternal lineage and 7.8% the Pietrain maternal lineage. BB was observed in 22.3% (6.8% were of the Large White paternal lineage, 9.7% of the Pietrain paternal lineage and 5.8% of the Pietrain maternal lineage). The allele frequencies were 49.0% A and 51.0% B allele. When we examined the Pietrain maternal and paternal lineages, we found that the PIT1 gene had been fixed in the paternal lineage, suggesting that the B allele is associated with low body fat and improved muscle development when compared to the maternal lineage. However, no significant differences were found between the Pietrain and Large White paternal lineages.

ABSTRACT. Genes whose products function in a common biological process are often co-regulated. When regulation occurs at the transcriptional level, co-expressed genes can be detected globally by expression arrays or by sequencing non-normalized cDNA libraries. We examined bovine gene expression in 27 tissues using non-normalized cDNA library sequencing. Contigs were generated from expressed sequence tags whose sequences overlapped. Contigs containing a minimum of five expressed sequence tags were ordered via a hierarchical clustering process, where the distance between the contigs represents their expression pattern similarity across tissues. Gene ontology terms associated with the genes in each cluster showed that co-clustered genes encoded proteins involved in a common biological process. This process can be used to annotate genes of unknown function in the cluster. Gene expression was compared between bovine and human tissues; there were significant correlations between species for each tissue, with the exception of thyroid and placenta. Tissues were also clustered based on the genes they express; tissues with similar physiological functions clustered closely. Based on this information, we generated the first preliminary gene atlas of the bovine genome. Genes with similar expression patterns were clustered, and genes with a common function co-clustered. This method can be used to annotate genes of unknown function in the bovine genome.

ABSTRACT. There are many candidate genes for chronic obstructive pulmonary disease (COPD). One such candidate is the group of genes that code for matrix metalloproteinases (MMPs), which play an essential role in tissue remodeling and repair associated with COPD. We tested the hypothesis that polymorphic variation in MMP genes influences the risk of developing COPD by examining functional polymorphisms in the promoters of MMP-3, MMP-9 and MMP-12 genes in 111 COPD patients and 101 controls. The -1171 5A/6A MMP-3, -1562 C/T MMP-9 and -82 A/G MMP-12 polymorphisms were analyzed by polymerase chain reaction, followed by restriction digestion. No significant differences were observed in allele and genotype frequencies between COPD patients and controls. Haplotype analysis also did not reveal differences between COPD patients and controls. We found that MMP polymorphisms had no significant impact on the risk of developing COPD in this Brazilian sample.

ABSTRACT. Quantitative and qualitative relationships were found between secreted proteins and their activity, and the hyphal morphol­ogy of Moniliophthora perniciosa, the causal agent of witches’ broom disease in Theobroma cacao. This fungus was grown on fermentable and non-fermentable carbon sources; significant differences in myce­lial morphology were observed and correlated with the carbon source. A biological assay performed with Nicotiana tabacum leaves revealed that the necrosis-related activity of extracellular fungal proteins also differed with carbon source. There were clear differences in the type and quantity of the secreted proteins. In addition, the expression of the cacao molecular chaperone BiP increased after treatment with secreted proteins, suggesting a physiological response to the fungus secretome. We suggest that the carbon source-dependent energy metabolism of M. perniciosa results in physiological alterations in protein expression and secretion; these may affect not only M. perniciosa growth, but also its ability to express pathogenicity proteins.

ABSTRACT. Eight cassava (Manihot esculenta Crantz) cultivars, of which four are indigenous to the Amazon region, along with an interspecific hybrid, were assessed for their carotenoid content. They were propagated and are maintained in the living Manihot col­lection at the University of Brasília. The cultivar “Amarelinha do Amapá”, which was collected from the State of Amapá, was found to have a very high content of b-carotene: 27 mg per 100 g, which is more than 50-fold higher than in other cassava cultivars. This cultivar, which also has excellent palatability, could be an important source of b-carotene for cassava breeding.

ABSTRACT. Water resource degradation is one of mankind’s greatest worries, as it causes direct and indirect damage to the associated biota. We initiated a water monitoring study in Pelotas Creek in 2003 in order to assess the mutagenic effect of the creek’s waters. Allium cepa cells exposed to water samples and a chronically exposed macrophyte were analyzed, through evaluation of the mitotic index, mitotic anomalies, interphase anomalies, and total anomalies. Five points were chosen along the lower course of Pelotas Creek, from which water samples and floating pennywort (Hydrocotyle ranunculoides, Apiaceae) were col­lected in 2006 and 2007. The enteric bacterium Escherichia coli was found at all sampling points; in the physical-chemical analysis, a few variables exceeded permitted limits, pH (from 6 to 9), chloride (250 mg/L), hardness (from 10 to 200 mg CaCO3/L), and conductivity (100 μΩ/cm). There was an increased number of cytogenetic anomalies in exposed A. cepa cells and in the pennywort in 2006 relative to 2007, which may be explained by the increased rainfall, which was three times greater in 2007 at some stations than in 2006.

ABSTRACT. Cochliomyia hominivorax larvae are known for their parasitic habit in living vertebrates, causing considerable economic losses to livestock industry. This ectoparasite has been controlled mainly by applying insecticides, but this method usually results in the selection of resistant individuals. The resistance mechanism known as knockdown resistance (kdr) is a generic term for amino acid substitutions in the sodium channel associated with pyrethroid resistance, and substitutions in residue 251 of the carboxylesterase E3 have been associated with organophosphate and pyrethroid hydrolysis. We looked for L1014F kdr and W251S mutations in the sodium channel and E3 genes, respectively, in C. hominivorax populations. Ten populations obtained from its current distribution were investigated using the polymerase chain reaction-restriction fragment length polymorphism technique. No mutant individuals were found for the kdr mutation. However, the W251S mutation was found in all populations investigated. Results from a bioassay with cypermethrin (a pyrethroid) indicated that the survival at the lowest concentration (Fisher exact test, P = 0.0003) and an intermediate concentration (P = 0.024) were associated with the W251S mutation. This correlation was not significant at the highest concentration tested (P = 0.221). We suggest that the W251S mutation in the C. hominivorax carboxylesterase E3 is also associated with pyrethroid hydrolysis. This information may contribute to the elaboration of improved management programs for this ectoparasite.

ABSTRACT. Molossidae species, Cynomops abrasus (2n = 34, fun­damental number, FN = 64), Eumops auripendulus (2n = 42, FN = 62), Molossus rufus (2n = 48, FN = 64), Molossops temminckii (2n = 48, FN = 64), and Nyctinomops laticaudatus (2n = 48, FN = 64), and Phyl­lostomidae species, Phyllostomus discolor (2n = 32, FN = 60), have karyotypes with different chromosome and fundamental numbers, dif­ferent localization of constitutive heterochromatin, and different num­bers and location of nucleolar organizer regions (NORs). Fluorescence in situ hybridization with a human probe of the telomeric sequence (TTAGGG)_n produced fluorescent signals in telomeric regions of the six bat species’ chromosomes; in E. auripendulus, pericentromeric signals were also observed in the acrocentric and subtelocentric chro­mosomes. A relationship between telomeric sequences and NORs, and between telomeric sequences and constitutive heterochromatin was de­tected in chromosomes bearing NORs in C. abrasus, M. temminckii, N. laticaudatus, and P. discolor. No interstitial signal was observed in the meta- or submetacentric chromosomes of these species.

ABSTRACT. Tamarindus indica has been used in folk medicine as an antidiabetic, a digestive aid, and a carminative, among other uses. Currently, there is no information in the toxicology literature concerning the safety of T. indica extract. We evaluated the clastogenic and/or genotoxic potential of fruit pulp extract of this plant in vivo in peripheral blood and liver cells of Wistar rats, using the comet assay, and in bone marrow cells of Swiss mice, using the micronucleus test. The extract was administered by gavage at doses of 1000, 1500 and 2000 mg/kg body weight. Peripheral blood and liver cells from Wistar rats were collected 24 h after treatment, for the comet assay. The micronucleus test was carried out in bone marrow cells from Swiss mice collected 24 h after treatment. The extract made with T. indica was devoid of clastogenic and genotoxic activities in the cells of the rodents, when administered orally at these three acute doses.

ABSTRACT. Male infertility is a heterogeneous disorder, with vari­ous genetic and environmental factors that contribute to the impairment of spermatogenesis. Genetic causes are responsible for 60% of cases of idiopathic infertility. Polymorphisms of genes that encode detoxi­fying enzymes of phase II drug metabolism can modify their expres­sion or function, affecting the biotransformation of toxic compounds to which the male reproductive system is exposed. GSTM1 and GSTT1 encode enzymes that are essential in the process of detoxification of endogenous and exogenous xenobiotics, facilitating their excretion. We examined GSTM1 and GSTT1 polymorphism in 233 men with idiopathic infertility seen at the Human Reproduction Service of the Federal University of Goiás from 2004-2006. Genotype GSTM1/T1 (null) was found in 30/105 normal individuals and in 64/128 abnormal individuals, indicating a significant association with idiopathic male infertility. The sperm alteration associated with greater GSTM1/T1 (null) frequency was decreased sperm count (oligozoospermy), which was more frequent in patients with GSTM1 (78.9%) as well as in those who had GSTT1 (73.7%), although the difference was not significant. Individuals polymorphic for genes GSTM1 and GSTT1 are susceptible to reduction in sperm quality and infertility, possibly because oligozoo­ spermic individuals have been affected by GST polymorphism.

ABSTRACT. The Batrachoididae includes some venomous brackish and marine fish found in the Atlantic, Indian and Pacific oceans. This family is composed of 69 species, distributed among 19 genera. Species of the genus Thalassophryne have been reported along the coast of Rio Grande do Norte (Brazil); T. nattereri has been responsible for a large number of human injuries. Little is known about the cytogenetic features of this family. We made a karyotypic characterization of T. nattereri col­lected from the estuary of the Apodi/Mossoró River, using conventional Giemsa staining, C-banding and silver nitrate-nucleolar organizer region technique. There was a modal diploid value of 2n = 46 chromosomes (8m + 8sm + 24st + 6a; fundamental number = 86). Single ribosomal sites were detected in the terminal region on short arms of a subtelocen­tric pair (19th). Heterochromatin segments were preferentially located over centromeric regions in some chromosome pairs. Pericentric inver­sions and Robertsonian rearrangements seem to have played a major role in karyotype evolution within this genus of toadfish.

ABSTRACT. The fig tree (Ficus carica L.) is of significant socio-economic importance in Turkey, with 25% of the world’s fig production. Genetic variation and relationships among 14 wild-grown figs sampled from Coruh Valley in Turkey were characterized by random amplified polymorphic DNA (RAPD). Ninety-eight DNA fragments were scored after amplification of DNA samples with 13 random primers; 70% of the scored bands were polymorphic. Genetic distances between the fig genotypes ranged from 0.21 to 0.62. Genotypes 08-ART-02 and 08-ART-06 were found to be the most closely related, whereas 08-ART-09 and 08-ART-10 were the most distant. The 14 wild-grown genotypes were grouped into six main clusters and one outgroup. We conclude that RAPD analysis is efficient for genotyping wild-grown fig genotypes.

ABSTRACT. In early development, female embryos (XX) produce twice the transcripts of X-linked genes compared with male embryos (XY). Dur­ing the course of development, inactivation of the X chromosome equili­brates gene dosage, making the development of female embryos viable. Moreover, the biotechnologies used for producing embryos in vitro seem to work better with male embryos, making it easier for them to reach the blastocyst stage and allow for complete gestation. We investigated the ex­pression of three X-linked genes that are involved in development, XIST, G6PD, and HPRT, and of the transcript interferon-τ, in male and female bovine blastocysts produced by nuclear transfer (NT) and by in vitro fer­tilization (IVF). Oocytes that had been matured in vitro were enucleated and reconstructed with somatic cells from adult animals at 18 h post-mat­uration. After fusion (two pulses of 2.25 kv/cm) and chemical activation (5.0 mM ionomycin for 5 min and 2.0 mM 6-DMAP for 3 h), the oocyte-somatic cell units were cultivated in CR2 with a monolayer of granulosa cells at 38.8ºC, in a humidified 5% CO₂ atmosphere. IVF embryos were inseminated, after centrifugation in a Percoll gradient, with 2 x 10⁶ sperm/mL TALP medium supplemented with BSA and PHE and cultivated un­der the same conditions as the cloned embryos. We used real-time PCR to analyze the gene expression of individual blastocysts compared to expres­sion of the housekeeping gene, GAPDH. The gene XIST was expressed in female embryos and not in male embryos produced by IVF, though it was expressed at low levels in male embryos produced by NT. Unlike previous reports, we found lower levels of the transcript of G6PD in females than in males, suggesting double silencing or other mechanisms of control of this gene. Female embryos produced by IVF expressed the HPRT gene at a higher level than female embryos produced by NT, suggesting that gene silencing proceeds faster in NT-produced female embryos due to “inacti­vation memory” from the nucleus donor. In conclusion, male and female embryos express different levels of X-chromosome genes and failures of these genes that are essential for development could reduce the viability of females. Nuclear transfer can modify this relation, possibly due to epigen­etic memory, leading to frequent failures in nuclear reprogramming.

ABSTRACT. The phenotype of partial trisomy 9p includes global developmental delay, microcephaly, bulbous nose, downturned oral commissures, malformed ears, hypotonia, and severe cognitive and language disorders. We present a case report and a comparative review of clinical findings on this condition, focusing on speech-language development, cognitive abilities and swallowing evaluation. We suggest that oropharyngeal dysphagia should be further investigated, considering that pulmonary and nutritional disorders affect the survival and quality of life of the patient. As far as we know, this is the first study of a patient with partial trisomy 9p described with oropharyngeal dysphagia.

ABSTRACT. We examined the stem anatomy of Manihot esculenta Crantz and its hybrid with M. oligantha Pax. Cross-sections were stained with safranin-alcian blue. Manihot esculenta and its hybrid were found to have the same vascular structure in bicollateral bundles with internal phloem, which has been associated with drought resistance in other plant groups. If this association is established for cassava, it would facilitate the selection of lines that are more adapted to arid regions. This is the first report of internal phloem in this genus.

ABSTRACT. Low purification efficiency and incomplete characteriza­tion of male goat (buck) spermadhesins (Bdhs) prompted us to develop an effective system to produce recombinant Bdhs (rBdhs). Bdh-2 cDNA was inserted into a prokaryotic expression plasmid, pTrcHis TOPO. The pTrcHis-Bdh-2 system was constructed to produce a His6 fusion protein in Escherichia coli Top10 cells. Recombinant clones were se­lected by growth in ampicillin-enriched medium, PCR amplification and nucleotide sequencing. The inserted cDNA was completely identi­fied and recombinant protein synthesis was monitored by SDS-PAGE, followed by immunoblotting with monoclonal anti-His antibody. Ex­pression of insoluble rBdh-2 was achieved at 0.1 to 2.0 mM IPTG, after 2 to 6 h of induction. Significantly increased production of rBdh-2 (P < 0.01) occurred with 1.5 mM IPTG after 2 h of induction, and with 0.3 mM IPTG after 4 h in culture. Among the induction times investigated, a period of 6 h gave the lowest levels of rBdh-2 production; with a 6-h incubation, there were no significant differences in rBdh-2 production for the various concentrations of IPTG tested (P > 0.05). The apparent molecular weight of rBdh-2 was 15.85 ± 0.09 kDa, calculated by im­age analysis of membranes. This is similar to the theoretical molecu­lar weight of 15.5 kDa predicted from the nucleotide sequence. Prior to this study, expression of recombinant goat spermadhesin had never been reported. Thus, an effective prokaryotic rBdh-2 expression system was developed in order to provide an adequate tool for studying bio­functions of goat spermadhesins.

ABSTRACT. Genetic variation provides a basis upon which populations can be genetically improved. Management of animal genetic resources in order to minimize loss of genetic diversity both within and across breeds has recently received attention at different levels, e.g., breed, national and international levels. A major need for sustainable improvement and conservation programs is accurate estimates of population parameters, such as rate of inbreeding and effective population size. A software system (POPREP) is presented that automatically generates a typeset report. Key parameters for population management, such as age structure, generation interval, variance in family size, rate of inbreeding, and effective population size form the core part of this report. The report includes a default text that describes definition, computation and meaning of the various parameters. The report is summarized in two pdf files, named Population Structure and Pedigree Analysis Reports. In addition, results (e.g., individual inbreeding coefficients, rate of inbreeding and effective population size) are stored in comma-separate-values files that are available for further processing. Pedigree data from eight livestock breeds from different species and countries were used to describe the potential of POPREP and to highlight areas for further research.

ABSTRACT. In Colombia, the mosquito Ochlerotatus taeniorhyn­chus has been identified as an efficient vector of the epidemic-epi­zootic Venezuelan equine encephalitis virus. We evaluated the ge­netic variability and heterogeneity of this mosquito in Colombian populations using eight microsatellite DNA loci. Two hundred and ten mosquito specimens collected from seven populations of the Co­lombian Atlantic coast (San Bernardo del Viento, Coveñas, Carta­gena, Barranquilla, Ciénaga, Dibulla, and Riohacha) were analyzed. We found five polymorphic microsatellite loci, with 19 alleles giv­ing 62.5% polymorphism; the mean number of alleles per locus was 3.8. The mean expected heterogeneity ranged from 0.568 to 0.660. Most of the polymorphic microsatellite loci were in Hardy-Wein­berg disequilibrium, due to both deficit and excess of heterozygotes. The Fst statistic gave a total value of 0.0369, reflecting low ge­netic differentiation among the populations and, as a consequence, a low degree of structuring among them, while gene flow was high (Nm = 6.52); these findings point to genetic homogeneity among these populations. There was no significant linkage disequilibrium between genotype pairs of the various populations. We concluded that this mosquito is distributed in local populations along the Co­lombian Atlantic coast; these findings will be useful for developing strategies for controlling this vector.

ABSTRACT. Human cytomegalovirus (HCMV) genetic determinants of endothelial cell tropism, leukocytes and dendritic cells have been identified in the genes UL131A, UL130, and UL128. We examined the structure of these three genes in HCMV. Eighteen low-passage clinical isolates and five non-passage strains from congenitally HCMV-infected infants in China were used to assess the structures of the UL131A, UL130, and UL128 genes and to find possible relationships between sequence polymorphism and different signs of HCMV disease. Comparisons were made between the UL131A, UL130, and UL128 genes of clinical strains and published sequences of Towne and Merlin strains. The UL131A coding region in the clinical strains was similar to that of Towne and Merlin strains, while UL130, and UL128 coding regions in the clinical strains were parallel with those of Towne and Merlin, respectively. Sequence comparison indicated that the UL130, and UL128 genes encode chemokine-like proteins in the clinical strain; the transmembrane regions of UL131A, and UL130 were conserved in all clinical and reference strains. The three genes of clinical strains from infants with different signs of HCMV disease had similar structure characterization. We conclude that the UL131A, UL130, and UL128 genes are highly conserved in these clinical strains. No correlation was found between the structure of the three genes and variations in HCMV disease. The finding of chemokine-like domains in UL130, and UL128 putative proteins suggests that the predicted products play a role in HCMV infectivity.