ABSTRACT. Male infertility is a common barrier that prevents successful conception. There have been reports of azoospermia in men with familial Mediterranean fever, some of whom had not been treated with colchicine. Variation in this disorder could be a risk factor for amyloidosis associated with azoospermia. We determined the frequency of 6 of the most common Mediterranean fever gene mutations, M680I, M694V, M694I, V726A, P369S, and A744S, in 74 infertile men, 155 men diagnosed with familial Mediterranean fever and 55 healthy fertile men in eastern Turkey. All three groups were screened for the 6 mutations using an amplification refractory mutation system and restriction fragment length polymorphism methods. Allelic frequencies were 2.7% for M694V and 1.35% for V726A in the infertile patient group and 1.8% for M694V and 1.8% for V726A in healthy subjects. Other mutations were not detected in patients or controls. The mutation frequency was not found to be significantly higher in infertile patients when compared with healthy fertile male controls. To our knowledge, this is the first study to determine the frequency of Mediterranean fever gene mutations in infertile male and the infertility rate of male patients with familial Mediterranean fever.

ABSTRACT. Acid phosphatases (AcPs) are known to provide phosphate to tissues that have high energy requirements, especially during development, growth and maturation. During spermatogenesis AcP activity is manifested in heterophagous lysosomes of Sertoli cells. This phagocytic function appears to be hormone-independent. We examined the expression pattern of AcP during the reproductive period of four species belonging to different vertebrate groups: Tilapia rendalli (Teleostei, Cichlidae), Dendropsophus minutus (Amphibia, Anura), Meriones unguiculatus (Mammalia, Rodentia), and Oryctolagus cuniculus (Mammalia, Lagomorpha). To demonstrate AcP activity, cryosections were processed for enzyme histochemistry by a modification of the method of Gömöri. AcP activity was similar in the testes of these four species. Testes of T. rendalli, D. minutus and M. unguiculatus showed an intense reaction in the Sertoli cell region. AcP activity was detected in the testes of D. minutus and O. cuniculus in seminiferous epithelium regions, where cells are found in more advanced stages of development. The seminiferous epithelium of all four species exhibited AcP activity, mainly in the cytoplasm of either Sertoli cells or germ cells. These findings reinforce the importance of AcP activity during the spermatogenesis process in vertebrates.

ABSTRACT. Several polymorphisms in the DNA repair gene are thought to have significant effects on cancer risk. We investigated the association of polymorphisms in the DNA repair genes XRCC1 Arg399Gln, XRCC3 Thr241Met, XPD Lys751Gln, XPG Asp1104His, APE1 Asp148Glu, and HOGG1 Ser326Cys with endometriosis risk. Genotypes were determined by PCR-RFLP assays in 52 patients with endometriosis and 101 age-matched healthy controls. Although there were no significant (P > 0.05) differences in the frequencies of genotypes or alleles of APE1, XRCC1, XPD, XPG, and HOGG1 genes between patients and controls, the frequency of the XRCC3 Thr/Thr genotype was significantly greater in endometriosis patients compared with controls (P = 0.005). XRCC3 Thr/Met genotypes (P = 0.022), and the Met allele (P = 0.005) seem to have a protective role against endometriosis. The distributions of genotypes and alleles of the genes APE1, XRCC1, XRCC3, XPD, XPG, and HOGG1 were not significantly associated with the different stages of endometriosis (P > 0.05). We conclude that the XRCC3 Thr/Thr genotype is associated with endometriosis in Turkish women.

ABSTRACT. Transposable elements contribute to the size, structure, variation, and diversity of the genome and have major effects on gene function. Sequencing projects have revealed the diversity of transposable elements in many organisms and have shown that they constitute a high percentage of the genome. PCR-based techniques using degenerate primers designed from conserved enzyme domains of transposable elements can provide quick and extensive surveys, making study of diversity and abundance and their applications possible in species where full genome sequence data are not yet available. We studied cassava (Manihot esculenta) En/Spm-like transposons (Meens) with regard to genomic distribution, sequence diversity and methylation status. Cassava transposase fragments characteristic of En/Spm-like transposons were isolated, cloned and characterized. Sequence analysis showed that cassava En/Spm-like elements are highly conserved, with overall identity in the range of 68-98%. Southern hybridization supports the presence of multiple copies of En/Spm-like transposons integrated in the genome of all cassava cultivars that we tested. Hybridization patterns of HpaII- and MspI-digested cassava genomic DNA revealed highly methylated sequences. There were no clear differences in hybridization pattern between the cultivars. We did not detect RNA transcripts of Meens by Northern procedures. We examined the possibility of recent transposition activities of the cassava En/Spm-like elements.

ABSTRACT. Two TP53 gene polymorphisms at codon 47 (TP53 Pro47Ser) and at codon 72 (TP53 Arg72Pro) have been associated with susceptibility to various cancers. We carried out a case-control study and examined the genotype distribution of TP53 Pro47Ser and Arg72Pro single nucleotide polymorphisms (SNPs), using a PCR-RFLP approach, to determine if these two SNPs are risk factors for colorectal cancer (CRC) development and to look for a possible correlation of these two SNPs with clinicopathological variables of CRC. We investigated the genotype distribution of these SNPs in 86 CRC cases in comparison with 160 healthy subjects in an ethnic Kashmiri population. TP53 Arg72Pro SNP genotype frequencies differed significantly (P = 0.000001) between the groups; the frequency of the Pro/Pro mutant was almost 20% in the general population. We also found significant association of the Pro/Pro mutant with tumor location, nodal status/higher tumor grade and bleeding per rectum/constipation. We conclude that Arg72Pro SNP is associated with susceptibility to developing CRC in this ethnic Kashmiri population.

ABSTRACT. Colletotrichum gloeosporioides is an important pathogen for a great number of economically important crops. During the necrotrophic phase of infection by Colletotrichum spp, the degradative enzymes of plant cell walls, such as pectate lyase, clearly increase. A gene pelB that expresses a pectate lyase was identified in isolates of C. gloeosporioides in avocado pathogens. Various molecular studies have identified a kind of specialization of C. gloeosporioides isolates with specific hosts; however, there have been no studies of this gene in isolates from hosts other than avocado. The same is true for other species of Colletotrichum. We examined genetic variability in order to design primers that would amplify pelB gene fragments and compared the products of this amplification in C. gloeosporioides isolates from different hosts. Genetic variability was assessed using ISSR primers; the resultant data were grouped based on the UPGMA clustering method. Primers for the pelB gene were designed from selected GenBank sequences using the Primer 3 program at an annealing temperature of 60°C and product amplification of nearly 600 bp. The ISSR primers were efficient in demonstrating the genetic variability of the Colletotrichum isolates and in distinguishing C. gloeosporioides, C. acutatum and C. sublineolum species. The gene pelB was found in C. gloeosporioides, C. acutatum and C. sublineolum. Amplified restriction fragments using MspI did not reveal differences in pelB gene structure in isolates from the three different host species that we investigated.

ABSTRACT. Brycon is one of the main genera of Neotropical freshwater fish. In Brazil, Brycon species have been found in many hydrographic basins, such as the Amazon, Paraná, Paraguay, and Araguaia-Tocantins basins. We examined the phylogenetic relationships among the species Brycon orbignyanus, B. hilarii, B. cf. pesu, B. cephalus, B. falcatus, and B. gouldingi, using mitochondrial and nuclear molecular markers. Specimens of B. orbignyanus were collected in the Paraná River. Specimens of B. hilarii were collected in the Manso River. Specimens of B. cephalus were obtained from a fish farm, and specimens of B. cf. pesu, B. falcatus and B. gouldingi were sampled in the Araguaia-Tocantins basin. DNA extraction was carried out using the phenol/chloroform method. Molecular polymorphism studies of Brycon species were carried out with the inter-simple sequence repeat (ISSR) technique, using the total DNA of six specimens of each species. In DNA amplification of B. cf. pesu, eight specimens were used. The partial sequence of mitochondrial cytochrome b was amplified by PCR. The PCR products were used directly in sequencing reactions. Each ISSR primer produced from 7 to 14 scorable and reproducible bands. The (GGAC)3A and (GGAC)3C primers produced the greatest number of species-specific bands. A 264-bp fragment, corresponding to the partial region of mitochondrial DNA cytochrome b, was sequenced and used for analysis. According to the phylogenetic tree obtained from the data, these Brycon species can be divided into two clades: one comprised only B. cf. pesu, and the second comprised the remaining Brycon species. We conclude that ISSR primers can be used for the identification of species-specific bands in fish, such as Brycon spp.

ABSTRACT. Chemokines are potent proinflammatory cytokines that are implicated in numerous inflammatory diseases. Proinflammatory gene polymorphisms lead to variations in the production and concentration of inflammatory proteins. We investigated a possible association between polymorphisms in chemokine and chemokine receptor genes (MCP-1 A-2518G and CCR2-V64I) and bladder cancer risk. Genotypes were determined by PCR-RFLP assays in 72 bladder cancer patients and 76 unrelated age-matched healthy controls. There were significant differences in the frequencies of the MCP-1 A-2518G (P = 0.012) and CCR2-V64I genotypes (P = 0.004) between the controls and patients. The MCP-1 A-2518G GG genotype frequencies for controls and cases were 0.039 and 0.11, respectively; individuals who had the GG genotype had a 3-fold increased risk of bladder cancer (P = 0.08). The CCR2-64I/64I genotype frequencies for controls and cases were 0.02 and 0.13, respectively; subjects carrying the 64I/64I genotype had a 5.9-fold increased risk of bladder cancer compared to the other genotypes. Individuals carrying the CCR2-V64I heterozygote or homozygous variant genotype (64I/64I + wt/64I) had a 2.9-fold increased risk of bladder cancer compared with the wild-type genotype (wt/wt). CCR2-V64I heterozygote or homozygous wild-type genotype (wt/wt + wt/64I) frequencies were significantly decreased in the patient group compared with controls. We conclude that CCR2-64I is a new risk factor for bladder cancer.

ABSTRACT. Palicourea coriacea (Rubiaceae) is a herbaceous, perennial species typical of the Cerrado; it is popularly known as “douradinha”, because of its yellow flowers. It is utilized in popular medicine, mainly for the treatment of renal diseases. We used RAPD markers to evaluate the genetic structure of nine natural populations of P. coriacea, totaling 168 individuals, collected in the States of Goiás and Bahia. This species showed a high level of genetic diversity, with He values varying between 0.259 and 0.338, with an overall mean of 0.296. Analysis by AMOVA revealed that 23% of the total variability was between populations and 77% was within populations. The estimate of apparent gene flow (Nm) was 0.83. Analyses of the fixation index (f ) using a Bayesian approach yielded a mean value of 0.98, suggesting that this is an autogamous species. Analyses of genetic divergence and spatial pattern of the populations utilizing θ^B and Φ_ST matrices, pair to pair, revealed no correlation between geographic distance and genetic distance; the nine populations grouped randomly, without relation to their geographic origin. The hypothesis that geographically close populations should be genetically close was discarded based on the Mantel test; the correlation was 0.155 (P = 0.23). The degree of interpopulational differentiation was relatively high, which allows us to recommend a strategy of sampling for the ex situ conservation of genetic variability, utilizing a larger number of populations. For in situ conservation, we suggest preservation of a larger number of areas in the Cerrado, where this species naturally occurs.

ABSTRACT. The narrow genetic base of soybean makes cultivar characterization based on morphological descriptors difficult; this characterization is mainly done for registration and protection. Correct characterization of cultivars could be achieved through molecular markers, since the frequencies of each allele in the population are known. Consequently, we developed a molecular characterization method and initiated the construction of a molecular database for soybean cultivar identification. Thirty-two soybean cultivars were analyzed with 48 fluorescent-labeled microsatellite markers. The reactions were carried out in singleplex, and genotyping in quadriplex, using a capillary electrophoresis system in an automated sequencer. Probabilities of random identity and probabilities of random identity exclusion were calculated through estimated allele frequencies. A characterization profile was considered when the probability of random identity exclusion was equal or superior to 99.9999%. All steps of the experiment were doubled, using two independent sets of the same cultivar to evaluate the reproducibility of the method. A set of 13 microsatellite markers identified all 32 cultivars with 99.9999% certainty. The method was efficient and precise, with high reproducibility for cultivar characterization. These data are the beginning of a molecular database for soybean, and they can be used for cultivar characterization for registration and protection purposes and for cultivar identification in cases of intellectual property enforcement.

ABSTRACT. Complete blood counts and hemoglobin isoform data were gathered from 36 specimens of the turtle species Phrynops geof froanus from the northwestern region of São Paulo State, Brazil. They were collected in an urban area. The hemoglobin profiles were obtained after red blood cell lysis and by electrophoretic migration in alkaline pH, acid pH, and neutral pH buffer. The hemoglobin components were confirmed using high-performance liquid chromatography (HPLC). Erythrogram analysis included hematocrit, total hemoglobin concentration, total red blood cell count, and red blood cell indices. The leukogram included a total white blood cell count and a calculation of the percent values of neutrophils, lymphocytes, monocytes, basophils, eosinophils, heterophils, and azurophils. HPLC analysis revealed three hemoglobin components; the first with a concentration of 5.5%, the second was a major component with an average concentration of 67.1%, and the third with a concentration of 28.5%. The hematological profile obtained for these specimens allowed us to establish a pattern for P. geoffroanus in São Paulo State Northwestern region. The average hematocrit values were 22.5% for females and 24.0% for males. For total hemoglobin, we found average values of 6.66 g/dL in females and 7.22 g/dL in males. The number of white blood cells was 2725 x 10³/μL for females and 2775 x 10³/μL for males. There was a predominance of heterophils, eosinophils, and monocytes in both sexes. No significant differences were found between males and females for hematological profile. The hematological results were compared to literature data for other Chelonia. They were similar to what is known for fresh water turtles.

ABSTRACT. We compared carcass and meat quality of pigs from the same sire line and two different dam lines, one that included Chinese breeds and one that did not. Line A consisted of 1/4 Landrace, 1/2 Large White, 1/8 Chinese breeds (Meishan, Fengjing, Jiaxing), and 1/8 Large White, Duroc and Pietrain, and line B consisted of 1/2 Large White and 1/2 Pietrain. The animals (N = 144) were slaughtered at a live weight of 108 kg. Backfat thickness, percentage of lean meat, pH 24 h after slaughter, meat color, percentage of drip loss, and percentage of intramuscular fat were measured and compared using analysis of variance in a completely randomized design; the BioEstat 5.0 test was applied for the comparison of means at a significance level of 5% for all analyses. Backfat was significantly lower for line A (12.78 mm) than for line B (15.90 mm). The pH measured 24 h after slaughter was significantly lower in line A (5.68) compared to line B (5.84). Percent lean meat was significantly higher for line A (61.21%) compared to line B (59.72%). Percentage drip loss was significantly higher in line A (2.73%) than in line B (2.23%). Percentage intramuscular fat and meat color were not significantly different between the lines. The inclusion of Chinese breeds produced a higher percentage of lean meat and reduced fat thickness, along with increased heterosis, which are important characteristics for breeding programs.

ABSTRACT. Angiotensin-converting enzyme (ACE) is a vital enzyme in the renin-angiotensin-aldosterone system, and there are reports in the literature describing its role in the development of cardiovascular system diseases, with I/D polymorphism of the ACE gene. We examined the relationship between a patient group with obstructive sleep apnea syndrome (OSAS) and a control group in terms of I/D polymorphism of the ACE gene. We examined 64 patients, with 37 individuals serving as the control group. PCR was used to detect ACE I/D gene polymorphism. Genotype was determined according to the bands that formed on agarose gel electrophoresis. Among the 64 OSAS patients, 27 were identified with the ID genotype, 27 with the DD genotype and 10 with the II genotype; among the 37 control subjects, 19 were identified with the ID genotype, 11 with the DD genotype and 7 with the II genotype. When the case group and controls were compared in terms of ID, II and DD genotypes, no significant difference was observed. On the other hand, when the two groups were compared with respect to mean body mass index, the OSAS group was found to be significantly different from the control group (P = 0.009). We conclude that ACE I/D gene polymorphism is not a genetic risk factor for OSAS in Turkish patients.

ABSTRACT. Acledra comprises 15 taxonomically identified species, most of which are crop pests. This is the first cytogenetic study of species of this genus. Acledra kinbergii and A. modesta showed the modal number of the Pentatomidae (2n = 14 = 12 + XY), while A. bonariensis had a reduced complement (2n = 12 = 10 + XY), with a markedly larger autosomal pair. Meiotic behavior follows the general pattern of the family; the autosomes divide pre-reductionally, the sex chromosomes are achiasmatic and divide post-reductionally, and at metaphase II the autosomes show a ring-shaped configuration with the pseudobivalent at the center. However, the configuration at metaphase I varies; A. modesta shows the typical arrangement (ring of bivalents with the sex chromosomes lying at its center). In A. kinbergii, the sex chromosomes are part of the ring or only the Y chromosome is at the center. In A. bonariensis, the ring arrangement is not well defined. There are also differences at the diffuse stage; chromatin strands of different width are observed in A. bonariensis and A. modesta, whereas bivalents do not entirely lose their identity in A. kinbergii. In A. bonariensis, the reduced complement may have originated from the fusion of the two larger non-homologous autosomes, which could characterize the ancestral karyotype of this genus. The presence of secondary constrictions in the larger pair of A. modesta and A. bonariensis may support this hypothesis. Since secondary constrictions are uncommon in the holokinetic chromosomes of heteropterans, their presence in these species may indicate that it is a plesiomorphic character of the genus.

ABSTRACT. The primary function of the human leukocyte antigen (HLA) system is to regulate the immune response. Because of its important role in the immune response and its high degree of polymorphism, the HLA system is associated with many diseases. We examined the polymorphisms of HLA-A, B and DRB1 alleles in 100 unrelated patients with lung carcinoma and in 438 unrelated normal controls of Han nationality from North China, using sequence-based typing and PCR with sequence-specific primers. We found that the frequencies of HLA-A*0201, A*2601, B*1518, B*3802, DRB1*0401, DRB1*0402, and DRB1*1201 were higher in the lung carcinoma group than in the normal control group. The P values were 0.035, 0.040, 0.001, 0.017, 0.014, 0.004, and 0.019, respectively, and the odds ratio values were 1.052, 3.513, 4.047, 3.054, 4.237, 19.397, and 2.128, respectively. The frequency of HLA-DRB1*1302 was lower in the lung carcinoma group than in the normal control group (P = 0.046, odds ratio = 0.168). We concluded that patients with lung cancer and healthy controls of Han nationality from North China differ in the frequencies of various HLA alleles.

ABSTRACT. Sorghum ranks fifth in worldwide economic importance among cereal crops and is one of the most important summer annual grasses of Pakistan. As it is a very diverse crop, sorghum genetic fingerprinting requires an efficient marker system. We estimated genetic divergence among 29 sorghum (Sorghum bicolor) genotypes, including approved varieties and local and exotic lines collected from different ecological regions of Pakistan, using random amplified polymorphic DNA (RAPD) markers. A total of 125 RAPD loci, with an average of 66 loci per genotype, were used to calculate genetic divergence among these genotypes, of which 119 were polymorphic, showing 95% overall polymorphism. Genetic similarity ranged from 0.36 to 0.92, indicating a relatively broad genetic base. RAPD analysis revealed maximum similarity between the Indian III and K-A-113 sorghum genotypes (both exotic lines), while the F-601 and F-606 were observed to be the most diverse genotypes. Mean band frequency revealed by these RAPD primers ranged from 0.17 to 0.56, with an average of 0.36. The data presented here support the findings that RAPDs can be effectively used for studying genetic diversity in sorghum.

ABSTRACT. Monocyte chemoattractant protein 1 (MCP-1) is an important chemokine that has a dose-dependent anti-tumoral effect. Polymorphism in the MCP-1 distal regulatory region (-2518A/G) can affect the level of MCP-1 expression. We examined the polymorphisms of 112 unrelated patients with non-small-cell lung cancer (NSCLC) and 82 unrelated healthy controls of Han nationality in North China using PCR-RFLP. We found that the distributions of AA, AG and GG genotypes of MCP-1-2518 were significantly different in NSCLC patients compared to controls (X² = 10.106, P = 0.006). There was a significant increase in the frequency of the AA genotype (odds ratio (OR) = 3.138, X² = 8.905, P = 0.003) and a significant decrease in the frequency of the GG genotype (OR = 0.516, X² = 4.613, P = 0.032) in the NSCLC patients, compared to controls. The frequencies of AA, AG and GG genotypes did not differ in the NSCLC patients according to the number of pack-years smoked. Based on these results, we suggest that the MCP-1 -2518A/G polymorphism is associated with genetic susceptibility to NSCLC.

ABSTRACT. We examined the efficiency of direct sequencing of pooled DNA for developing common single nucleotide polymorphisms (SNPs) and its accuracy for estimating allele frequencies. A pool of 200 control DNAs was established and was used for developing SNPs and estimating minor allele frequencies (MAF). The sensitivity of the pooled DNA method for successfully detecting an SNP with an MAF >0.01 listed in the database was approximately 0.7; it was particularly efficient for detecting SNPs with MAF >0.1, which is compatible with the common disease/common variant hypothesis. The mean difference between the estimated and the observed MAFs was 0.03 ± 0.023. The pooled DNA method identified four additional SNPs, for which the allele frequency information was not available in the database. The pooled DNA method is a cost- and time-effective tool for both qualifying and quantifying SNPs with considerable accuracy, and it can be particularly useful for dissecting the common disease/common variant hypothesis; this represents a best-case scenario for large-scale association mapping.

ABSTRACT. We report on a 23-year-old girl with short stature, short and wide neck, low posterior hairline, hypogonadism, underdeveloped breasts, infantile uterus, ovaries not visualized, and primary amenorrhea. Cytogenetic G-banding analysis revealed a mosaic karyotype of 46,X,dup(X)(q22)[35]/45,X[15], confirming the clinical suspicion of Turner syndrome. Molecular cytogenetics using a multicolor banding probe set for the X-chromosome characterized an inverted dup(X). The karyotype of the patient was therefore interpreted as 46,X,inv dup(X) (pter → q22::q22 → pter). This patient had a mosaic Turner syndrome with a cell line comprising partial trisomy Xpter to Xq22 and partial monosomy Xq22 to Xqter.

ABSTRACT. The relationship between pollen germination and the dynamic organization of the actin cytoskeleton during pollen germination is a central theme in plant reproductive biology research. Maize (Zea mays) pollen grains were implanted with 30 keV argon ion (Ar⁺) beams at doses ranging from 0.78 × 10¹⁵ to 13 × 10¹⁵ ions/cm². The effects of low-energy ion implantation on pollen germination viability and the dynamic organization of the actin cytoskeleton during pollen germination were studied using confocal laser scanning microscopy. Maize pollen germination rate increased remarkably with Ar⁺ dose, in the range from 3.9 × 1015 to 6.5 × 10¹⁵ ions/cm²; the germination rate peaked at an Ar⁺ dose of 5.2 × 10¹⁵ ions/cm². When the implantation dose exceeded 7.8 × 1015 ions/cm², the rate of pollen germination decreased sharply. The actin filaments assembled in pollen grains implanted with 5.2 × 10¹⁵ ions/cm² Ar⁺ much earlier than in controls. The actin filaments organized as longer parallel bundles and extended into the emerging pollen tube in treated pollen grains, while they formed random and loose fine bundles and were gathered at the pollen aperture in the control. The reorganization of actin cytoskeleton in the pollen implanted with 9.1 × 10¹⁵ ions/cm² Ar⁺ was slower than in controls. There was a positive correlation between pollen germination and the dynamic organization of the actin cytoskeleton during pollen germination. Ion implantation into pollen did not cause changes in the polarization of actin filaments and organelle dynamics in the pollen tubes. The effects of Ar⁺ implantation on pollen germination could be mediated by changes in the polymerization and rearrangement of actin polymers.

ABSTRACT. The tree species Parapiptadenia rigida, native to southern South America, is frequently used in reforestation of riverbanks in Brazil. This tree is also a source of gums, tannins and essential oils, and it has some medicinal uses. We investigated flooding tolerance and genetic diversity in two populations of P. rigida; one of them was naturally exposed to flooding. Plants derived from seeds collected from each population were submitted to variable periods of experimental waterlogging and submergence. Waterlogging promoted a decrease in biomass and structural adjustments, such as superficial roots with aerenchyma and hypertrophied lenticels, that contribute to increase atmospheric oxygen intake. Plants that were submerged had an even greater reduction in biomass and a high mortality rate (40%). The two populations varied significantly in their RAPD marker profiles, in their ability to produce aerenchyma when waterlogged and to survive when submerged, suggesting ecotypic differentiation between them. Hence, the seasonal flooding that has been challenging the tropical riparian forest appears to be genetically modifying the P. rigida populations exposed to it by selecting individuals with increased ability to live under this condition.

ABSTRACT. Amplified fragment length polymorphism (AFLP) technique was used to assess genetic relationships among 20 grapevine rootstocks in Turkey. Discrimination of the rootstocks with 10 primer combinations yielded 1366 bands on AFLP gels; 65% of them were polymorphic. The rootstocks revealed two main clusters; one of them comprised two (Malégue and Harmony) and the other 18 genotypes. The Ber x Rip hybrids Cosmo 2 and Cosmo 10 formed a group with a high internal similarity ratio (0.909); they also formed a group with other Ber x Rip hybrids, 5C, 8B, SO4, and 420A Mgt, with a similarity ratio higher than 0.690 (subcluster II). Rootstock 5BB was placed in another subcluster (subcluster III). Among five Ber x Rup rootstocks, 110R-99R (0.853) and 1103P-140Ru (0.837), which were located in different subclusters, formed a dual group, as expected. Rootstock 779P, which had almost 0.800 similarity with the dual group of 110R-99R, formed another group. The 44-53 Malégue and Harmony rootstocks formed a group with the lowest similarity ratio (0.668) (subcluster I) and 41B-Fercal formed another dual group with a high similarity ratio (0.813). The distinction capacity of single- and double-EcoRI-MseI primers was evaluated; primers AC/CTA, TC/CAC, AG/CTC, and AG/CAG discriminated the 20 rootstocks, with a similarity value below 0.910. The best primers for discrimination of rootstock varieties were AG/CAG and AG/CTC, while the primers TC/CAC and AC/CTA could also be useful for clonal discrimination of genotypes.

ABSTRACT. Abundant single nucleotide polymorphisms (SNPs) provide the most complete information for genome-wide association studies. However, due to the bottleneck of manual discovery of putative SNPs and the inaccessibility of the original sequencing reads, it is essential to develop a more efficient and accurate computational method for automated SNP detection. We propose a novel computational method to rapidly find true SNPs in public-available EST (expressed sequence tag) databases; this method is implemented as SNPDigger. EST sequences are clustered and aligned. SNP candidates are then obtained according to a measure of redundant frequency. Several new informative biological features, such as the structural neighbor profiles and the physical position of the SNP, were extracted from EST sequences, and the effectiveness of these features was demonstrated. An ensemble classifier, which employs a carefully selected feature set, was included for the imbalanced training data. The sensitivity and specificity of our method both exceeded 80% for human genetic data in the cross validation. Our method enables detection of SNPs from the user’s own EST dataset and can be used on species for which there is no genome data. Our tests showed that this method can effectively guide SNP discovery in ESTs and will be useful to avoid and save the cost of biological analyses.

ABSTRACT. We evaluated the efficiency of the touchdown method to determine the ideal PCR conditions for distinct inter-simple sequence repeat primers for processing DNA from common corn, popcorn, sweet corn, and a Tripsacum-maize hybrid. Genomic DNA was extracted from eight accessions of corn: two of the dent type, one Tripsacum-maize hybrid, one sweet corn, one flint-type corn, and three popcorn. Fifteen inter-simple sequence repeat primers were used: (CT)₈RC, (CT)₈TG, (GA)₈T, (GA)₈YC, (CTC)₅RC, (GTC)₆, (GA)₆CC, (GT)₆CC, (CAC)₃GC, (AG)₈YT, (AC)₈T, (AC)₈YG, (CT)₈RG, (GGAT)₃GA, and (GAA)₆AA. The annealing temperature and the melting temperature for each primer were estimated using a formula for RW Genes products, or we used the temperatures indicated by the manufacturer (Invitrogen). The touchdown method was then applied to each primer, varying the number of final cycles (10 or 12) and the decrease in temperature (0.5° or 1.0°C intervals). The gels were compared, considering the revelation quality, band sharpness and the number of bands visualized. The touchdown-PCR method was more efficient for band amplification for most of the primers, especially at higher annealing temperatures. This type of system is useful for reducing the resources, time and effort needed for optimizing temperature conditions for a group of representative primers.

ABSTRACT. The bovine transferrin gene (TF) is located at 125 cM on bovine chromosome 1 (BTA1); it codes for transferrin, a glycoprotein that is highly conserved in many species and that is responsible for iron transport. The TF gene has been located in several QTL regions, and some transferrin classes have been associated with fat and milk yields. We analyzed by means of allele-specific oligonucleotide real-time PCR the c.1455A>G SNP in exon 12 of the TF cDNA sequence (accession number U02564), which induces an Asp/Gly substitution at position 469 of the peptide. The c.1455A>G SNP was assayed in eight Spanish cattle breeds, as well as in two groups of Holstein-Friesian animals that had the highest and lowest estimated breeding values for milk fat yield. Analysis of the cSNP showed balanced frequencies in all breeds, with a mean of 0.44. Evaluation of a potential association between the cSNP and the groups of Holstein-Friesian animals selected for milk fat yield showed a significant association (P < 0.0006); the G allele was associated with high fat production. Significant differences in genotypic frequencies between the groups were also detected (P < 0.0028). These results lead us to suggest that the TF gene has an effect on milk fat yield.

ABSTRACT. Mariner-like elements are widely present in diverse organisms. These elements constitute a large fraction of the eukaryotic genome; they transpose by a “cut-and-paste” mechanism with their own transposase protein. We found two groups of mobile elements in the genus Rhynchosciara. PCR using primers designed from R. americana transposons (Ramar1 and Ramar2) were the starting point for this comparative study. Genomic DNA templates of four species: R. hollaenderi, R. millerii, R. baschanti, and Rhynchosciara sp were used and genomic sequences were amplified, sequenced and the molecular structures of the elements characterized as being putative mariner-like elements. The first group included the putative full-length elements. The second group was composed of defective mariner elements that contain a deletion overlapping most of the internal region of the transposase open reading frame. They were named Rmar1 (type 1) and Rmar2 (type 2), respectively. Many conserved amino acid blocks were identified, as well as a specific D,D(34)D signature motif that was defective in some elements. Based on predicted transposase sequences, these elements encode truncated proteins and are phylogenetically very close to mariner-like elements of the mauritiana subfamily. The inverted terminal repeat sequences that flanked the mariner-like elements are responsible for their mobility. These inverted terminal repeat sequences were identified by inverse PCR.

ABSTRACT. We determined the expression levels of DREB transcription factor (Gmdreb1) and of the genes Gmgols, Gmpip1b, Gmereb, and Gmdefensin in drought-tolerant (MG/BR46-Conquista) and drought-sensitive (BR16) genotypes of soybean, during drought. The trial was carried out in a controlled-environment chamber, set up to provide drought conditions. Sequences of Arabidopsis thaliana DREB-family proteins were used to build a phylogenetic tree through the alignment of the conserved regions near the AP2 domain. We found that Gmdreb1 is similar to Atrap2.1, which is located near the AtDREB1 and AtDREB2 families. The amplified fragment was cloned and sequenced; alignment with the sequence available at Genbank showed total similarity. Expression analysis showed that under drought: a) Gmdreb1 expression increased in leaves and roots of both genotypes and expression level changes occurred that were correlated with the length of the water-deficit period; b) there were increased expression levels of Gmdefensin in roots of MG/BR46; c) expression of Gmgols increased in leaves and roots of the two genotypes; d) Gmpip1b expression generally increased, except in roots of BR16, and e) the same was found for Gmereb, except in roots of MG/BR46.

ABSTRACT. We examined the variation of the BMP4 gene in four Chinese indigenous cattle breeds and investigated the association of this polymorphism with body measurement traits. Using PCR-SSCP and DNA sequencing, a polymorphic microsatellite was detected in the third exon of the bovine BMP4 gene in 459 samples from four Chinese indigenous cattle breeds, Qinchuan, Luxi, Nanyang, and Jiaxian red. The two alleles were named A and B. Allele frequencies of BMP4-A/B in the four breeds were 0.939/0.061, 0.928/0.072, 0.929/0.071, and 0.938/0.062, respectively. Least squares analysis revealed significant effects of genotype on withers height in the four breeds, on hip height in two breeds (Luxi and Nanyang, P < 0.05) and on chest circumference in Qinchuan (P < 0.05), while no significant effects of genotype on body length and rump length were found. These results can be applied to marker-assisted selection of Chinese cattle breeds, but a much larger number of animals will be needed for association analysis.

ABSTRACT. The IGF1 gene (insulin-like growth factor 1) is a candidate gene for marker-assisted selection strategies. A single nucleotide polymorphism in the promoter region (IGF1/SnaBI) has been reported to be associated with production traits in several cattle breeds. Here, we report its allelic frequencies in Charolais and Beefmaster breeds; we confirm its association with three growth traits: weaning weight, weaning weight adjusted to 210 days and preweaning weight gain in the Charolais breed. In addition, we designed a strategy to search these breeds for new polymorphisms in four coding regions of the gene. A C/A transversion was detected in intron 4, but it was not associated with the growth traits. A single nucleotide polymorphism (IGF1/SnaBI) is proposed as a selection marker for Mexican Charolais cattle; validation of its association with weaning weight, weaning weight adjusted to 210 days and preweaning weight gain, could complement the genetic evaluations of this breed through marker-assisted management strategies.

ABSTRACT. The RAPD technique was used for determining genetic differences between 12 wild-olive varieties grown in the Aegean provinces of Izmir, Mugla, and Manisa in Turkey. Wild olives obtained from the same provinces were included in the same plot. Twenty of 25 operon primers (OP-I 4, OP-I 14, OP-I 15, OP-I 16, OP-I 17, OP-Q1, OP-Q2, OP-Q3, OP-Q4, OP-Q11, OP-Q12, OP-Q13, OP-Q14, OP-Q15, OP-Q16, OP-Q17, OP-Q18, OP-Q19, OP-Q20, OP-F1, OP-F2, OP-F3, OP-F6, OP-F7, OP-F8) yielded bands. The differences between the varieties were determined based on their genetic similarities, using principal coordinate analysis; genetic distances were determined using neighbor-joining analysis. The varieties wild 7 and wild 12 had the lowest genetic similarity (0.97, Jaccard similarity index); they also had the greatest genetic distance between them (0.3606, Nei’s genetic distance). It was concluded that the RAPD technique is adequate for the evaluation of genetic relationships among wild olives. Principal coordinate analysis and neighbor-joining analysis gave results that support the use of this type of analysis to help understand the genetic background of olives and for further genetic studies.

ABSTRACT. Heat stress produces oxidative stress and affects the alternation of plasma K⁺ and Na⁺. Since Na⁺,K⁺-ATPase is sensitive to oxidative stress and critical for maintaining the homeostasis of these two ions, we examined the genetic polymorphism of the ATP1A1 gene in 160 Holstein cows using polymerase chain reaction low ionic strength single-strand conformation polymorphism and DNA sequencing methods. G to A at position -14103 in exon 14 and C to T at position -14242 in intron 14 of the bovine ATP1A1 gene were identified, but the former single nucleotide polymorphism was silent with respect to the amino acid sequence of the protein. However, we found significant correlations between ATP1A1 gene polymorphism and the coefficient of heat tolerance (P 0.01) and with respiratory rate (P 0.01). Genotype AC was the most favorable genotype for heat tolerance. This polymorphism site has potential as a genetic marker for heat tolerance traits in dairy cattle breeding.

ABSTRACT. Chronic obstructive pulmonary disease (COPD) is a multifactorial disease with possible genetic predisposition and involvement of various environmental factors. Several candidate genes have been reported as potentially associated with this lung disease. The glutathione S-transferase P1 gene (GSTP1) was proposed to be involved in susceptibility to develop COPD. It belongs to the GST family, which is a group of phase II enzymes that catalyze the glutathione conjugation of many endogenous and exogenous electrophilic compounds, such as carcinogens, therapeutic drugs, environmental toxins, and oxidative stress products. We conducted a case-control study to investigate genetic polymorphisms of this enzyme [exon 5 (Ile105Val) and exon 6 (Ala114Val)] in 234 unrelated COPD cases and 182 healthy controls from a Tunisian population. Genotyping was carried out using polymerase chain reaction and restriction fragment length polymorphism methods. GSTP1 Ala114/Val114 and Val114/Val114 genotypes were not found in either patients or healthy controls. However, there were differences in the distribution of various exon 5 GSTP1 genotypes between COPD patients and healthy controls. GSTP1 Val105/Val105 was significantly more common in patients compared to controls (OR = 2.67; 95%CI = 1.45-4.92; P = 0.0013). Multivariate logistic regression analysis confirmed a significant relationship between the mutant genotype and COPD (OR = 2.58; 95%CI = 1.31-5.09; P = 0.026), after adjustment for classic risk factors. Analysis of variance showed no correlation between age, body-mass index, pack-years, percentage of predicted FEV1 values, and any of the GSTP1 genotypes. We conclude that subjects with GSTP1 Val105 allele are at higher risk of COPD.

ABSTRACT. The effect of genetic and non-genetic factors for carcass, breast meat and leg weights, and yields of a commercial broiler line were investigated using the restricted maximum likelihood method, considering four different animal models, including or excluding maternal genetic effect with covariance between direct and maternal genetic effects, and maternal permanent environmental effect. The likelihood ratio test was used to determine the most adequate model for each trait. For carcass, breast, and leg weight, and for carcass and breast yield, maternal genetic and permanent environmental effects as well as the covariance between direct and maternal genetic effects were significant. The estimates of direct and maternal heritability were 0.17 and 0.04 for carcass weight, 0.26 and 0.06 for breast weight, 0.22 and 0.02 for leg weight, 0.32 and 0.02 for carcass yield, and 0.52 and 0.04 for breast yield, respectively. For leg yield, maternal permanent environmental effect was important, in addition to direct genetic effects. For that trait, direct heritability and maternal permanent environmental variance as a proportion of the phenotypic variance were 0.43 and 0.02, respectively. The results indicate that ignoring maternal effects in the models, even though they were of small magnitude (0.02 to 0.06), tended to overestimate direct genetic variance and heritability for all traits.

ABSTRACT. A molecular maker for authenticating species origin of the stingless bee (Trigona collina) was developed. Initially, amplified fragment length polymorphism analysis was made of 11 stingless bee species using 64 primer combinations. A 316-bp band found only in T. collina was cloned and sequenced. A primer pair (CUTc1-F/R) was designed and tested for species-specificity in 15 stingless bee species (239 nests). The expected 259-bp fragment was consistently amplified in all T. collina individuals (134/134 nests, 100%). Cross-species amplification was observed in T. pagdeni (43/51 nests; 84.3%), but not in other species. SSCP analysis of CUTc1 unambiguously differentiated T. collina from T. pagdeni. CUTc1 generated three genotypes in Thai T. collina (134 nests). An AA (259/259 bp) genotype was found in all stingless bees from the north (21 nests) and northeast (32 nests), and 23/28 nests from the Central region, whereas a BB (253/253 bp) genotype was observed in most samples from peninsular Thailand (42/53 nests). Heterozygotes exhibiting the AB (253/259 bp) genotype were observed in 5 of 28 nests from Prachuap Khiri Khan located slightly above the Kra ecotone and 11 of 53 nests originated further south of the Kra ecotone. Genotype distribution patterns of CUTc1 clearly indicated intraspecific population differentiation of Thai T. collina.

ABSTRACT. Twelve polymorphic microsatellites from the (AG)₁₃ and (CA)₁₃ enriched genomic libraries of Miichthys miiuy were isolated and characterized in a test population; the number of alleles ranged from two to nine. The observed and expected heterozygosities ranged from 0.1923 to 1.0000 and from 0.2633 to 0.8337, respectively. Three loci deviated from Hardy-Weinberg equilibrium, and linkage disequilibrium between five pairs of loci was significant. These polymorphic microsatellite loci can be used for genetic diversity analysis and molecular-assisted breeding of M. miiuy.

ABSTRACT. We developed a straightforward, rapid, and inexpensive method to determine transgene copy number in tobacco. The plasmid (pSSRCopy) used for tobacco transformation contains a simple sequence repeat (SSR) locus, PT1199, which was partially deleted in the middle, a homogenous SSR locus in tobacco K326. A 168-bp segment of the cloned PT1199 was shortened to 95 bp by deleting a 73-bp internal fragment. Using a pair of SSR primers, competitive PCR was amplified from genomic DNA from transgenic tobacco harboring pSSRCopy, and the two expected bands were found. The 168-bp band (SSR-168) corresponds to endogenous PT1199 and the 95-bp band (SSR-95) comes from the integrated pSSRCopy. A single copy of a transgene can be easily distinguished from multiple copies by comparing band densities.

ABSTRACT. Mutations in the EDA gene are responsible for X-linked hypohidrotic ectodermal dysplasia, the most common form of ectodermal dysplasia. Males show a severe form of this disease, while females often manifest mild to moderate symptoms. We identified a missense mutation (c.463C>T) in the EDA gene in a Jordanian family, using direct DNA sequencing. This mutation leads to an amino acid change of arginine to cysteine in the extracellular domain of ectodysplasin-A, a protein encoded by the EDA gene. The phenotype of a severely affected 11-year-old boy with this mutation included heat intolerance, sparse hair (hypotrichosis), absence of 17 teeth (oligodontia), speech problems, and damaged eccrine glands, resulting in reduced sweating (anhidrosis). Both the mother (40 years old) and the sister (10 years old) were carriers with mild to moderate symptoms of this disease, while the father was healthy. This detailed description of the phenotype caused by this missense mutation could be useful for prenatal diagnosis.

ABSTRACT. The Caenorhabditis elegans genome has several regular and irregular characteristics in its nucleotide composition; these are observed within and between chromosomes. To study these particularities, we carried out a multifractal analysis, which requires a large number of exponents to characterize scaling properties. We looked for a relationship between the genetic information content of the chromosomes and multifractal parameters and found less multifractality compared to the human genome. Differences in multifractality among chromosomes and in regions of chromosomes, and two group averages of chromosome regions were observed. All these differences were mainly dependent on differences in the contents of repetitive DNA. Based on these properties, we propose a nonlinear model for the structure of the C. elegans genome, with some biological implications. These results suggest that examining differences in multifractality is a viable approach for measuring local variations of genomic information contents along chromosomes. This approach could be extended to other genomes in order to characterize structural and functional regions of chromosomes.

ABSTRACT. Genetic similarities and distances between wild-type olives in Turkey were studied using an RAPD-PCR assay. Seven wild olive tree samples were collected from villages in Manisa and Izmir provinces. Genomic DNA was extracted from young leaves and the RAPD-PCR assay was used to generate RAPD markers. Sixty-five random primers obtained from Operon Technologies were tested for the assay (OP-A 1-20, OP-I 1-20, OP-Q 1-20, and OP-J 1-5). Thirty-two of these primers yielded 115 highly polymorphic bands. The mean number of usable bands per primer for all the samples was 3.59. The genetic distance values ranged from 0.1498 to 0.6845, and genetic similarity values varied from 0.8609 to 0.5043. We found that the closest samples based on their genetic distance and similarity values were from Harlak and Sabancilar; the most distant samples were from Bornova and Bademli.

ABSTRACT. he ethical aspects of the use of stored tissue samples collected from minors are of topical interest. However, the views of professionals working in the field of genetics have not been investigated in depth anywhere. We conducted a survey among 194 such professionals in Belgium. This list was composed of the members of the High Council for Anthropogenetics, supplemented with all professionals working in the field of genetics that we found on the websites of the eight Belgian centers of human genetics and of the associated university registries. We achieved a response rate of 35.5%. The vast majority (92%) think that research on stored tissue samples is useful. Most respondents stated that parental consent is valid (82.5%), and 76.5% thought that children should also be given the right to assent when they are able to comprehend the implications of the storage of biological samples and of genetic research. Slightly more than half put the age at which young people can understand storage or research rather high: 16-18 years (51 and 53.1%, respectively). Although there is some consensus in the literature that donors should be allowed to give broad consent for future research on their biological samples, only 47.6% in our survey thought that parents should be allowed to consent to any future research on their children’s samples. The aim of our study was to give some basis for future ethical reflections and policies on the subject of stored tissue samples from minors for genetic research. We concluded that a large majority of Belgian researchers and clinicians in the field of genetic research think research on stored tissue samples from minors is useful. They also think that parental consent for such research is valid, but that children should be allowed to assent as they grow older.

ABSTRACT. Turkey is one of the most important genetic resources of the date plum, Diospyros lotus, especially in the northeastern part of the country. Authenticating the identity of germplasm resources of D. lotus would be of great value for breeding. We examined the genetic variability of 11 D. lotus genotypes sampled from Coruh Valley in Turkey. One hundred and twenty-eight DNA markers were generated by 12 random primers. The highest polymorphism ratio was observed with the primer OPA-01 (71%) while the lowest was with OPY-01 (36%). The band size was between 350 and 2500 bp for these primers. The percentage of polymorphic bands was 58%, which demonstrated the efficiency of these primers. The similarity between genotypes ranged from 0.48 to 0.76. The RAPD markers permitted us to distinguish all the genotypes.

ABSTRACT. Turnip (Brassica rapa var. rapa) is one of the main vegetables consumed by people living in Eastern Anatolia in Turkey. In this region, farmers obtain their own seeds for production, which results in considerable morphological variability. We examined the genetic variation and relationships among 11 turnip genotypes sampled from diverse environments of the Erzurum region located in Eastern Anatolia in Turkey. Thirty-two Operon RAPD primers were screened; among them, 20 gave reproducible and clear DNA fragments after amplification. The average polymorphism ratio was 90.4%. The genetic distance between turnip genotypes were found to range from 0.302 to 0.733, indicating high genetic variability. Eleven genotypes were divided into three main clusters in a dendrogram; ETS2 and ETS8 genotypes were the most distant. We conclude that RAPD analysis would be useful for genotyping turnip genotypes.

ABSTRACT. Polygalacturonase-inhibiting proteins are leucine-rich repeat proteins that inhibit fungal endopolygalacturonases. The interaction of polygalacturonase-inhibiting protein with endopolygalacturonases limits the destructive potential of endopolygalacturonases and may trigger plant defense responses induced by oligogalacturonides. We examined the expression of fungal pg and plant Pvpgip genes in bean (Phaseolus vulgaris) stems infected with Sclerotinia sclerotiorum to determine whether any of them are associated with the infection process. Transcriptional analysis was carried out by means of semi-quantitative reverse transcription PCR or real-time PCR. The sspg1 gene was highly expressed during infection; sspg3 was regulated during the later phases of infection; sspg5 was more uniformly expressed during infection, whereas sspg6 was only weakly expressed. During the course of infection, Pvpgip1 transcripts were not detected at early stages, but they appeared 72 h post-inoculation. High levels of Pvpgip2 expression were observed during the initial phase of infection; the transcript peaked by 48 h post-inoculation and declined by 72 h post-inoculation. Pvpgip3 expression increased strongly at 96 h post-inoculation. Pvpgip4 was constantly present from 24 h post-inoculation until the end of the experiment. However, we detected higher levels of the Pvpgip4 transcript in the necrotic lesion area than in plants that had been mechanically wounded. Remarkably, only Pvpgip4 appeared to be moderately induced by mechanical wounding. These results provide evidence that endopolygalacturonases contribute to the infection process during host colonization by promoting the release of plant cell oligogalacturonides, which are powerful signaling molecules and may also activate plant defenses, such as polygalacturonase-inhibiting proteins.

ABSTRACT. Spittlebugs are the leading cause of damage to tall grasses. Annual losses are estimated to reach 2.1 billion dollars in sugarcane crops and grazing land throughout the world. Correct identification of these species is difficult due to similarities in color, body size and male genitalia. Molecular markers have been useful in the identification and assessment of genetic diversity of many species. We investigated the genetic diversity of the spittlebug species Mahanarva fimbriolata, M. spectabilis and M. liturata and looked for markers that could aid in their identification. DNA from 34 spittlebug specimens, collected from six different regions of Brazil (Brasília, Campo Grande, Valença, Presidente Prudente, Juiz de Fora, and Porto Alegre), was analyzed with 29 RAPD primers, generating 501 polymorphic markers. High genetic variability was found among individuals M. fimbriolata (0.37), M. spectabilis (0.18) and M. liturata (0.69). Species-specific molecular RAPD markers were identified for each of the three species; these could be used as auxiliary tools for their correct identification.

ABSTRACT. Forty genotypes (clones) of sugarcane, including elite lines, commercial cultivars of Saccharum officinarum and clones of S. barberi were fingerprinted with 50 SSR markers using a PCR-based marker assay. Nei’s genetic distances for SSR data were determined and relationships between accessions were portrayed graphically in the form of a dendrogram. Genetic distance values ranging from 0.60 to 1.11 were observed among the 40 sugarcane accessions. The shortest genetic distance of 0.60 was seen between genotypes US-804 and US-130. These two genotypes differed from each other only in 10 bands, with 20 primers. The most dissimilar of the accessions were CP-77-400 and US-133, with a genetic distance of 1.11. SSR fingerprints can help sugarcane breeders to clarify the genetic pedigree of commercial sugarcane varieties and evaluate the efficiency of breeding methods.

ABSTRACT. The stem structure of two cassava cultivars, UnB 99 and UnB 110, known for being adapted to humid conditions and tolerant to drought, respectively, and of a wild species, Manihot glaziovii, was examined anatomically. Free-hand sections of secondary stems were made, clarified with 50% sodium hypochlorite solution, stained with 1% alcian-blue safranin, and then passed through an ethanol series and butyl acetate, followed by mounting in synthetic resin. M. glaziovii stems had dense prismatic and druse crystals in the cortical parenchyma, along with abundant gelatinous fibers. The pericycle fibers also had thicker walls. An absence of crystals, offset by abundant starch, was observed in clone UnB 99. In M. glaziovii, abundant tyloses were found in vessel elements; these were rare in clones UnB 99 and UnB 110. The wild species had larger vascular vessels; the secondary xylem showed very little starch, unlike UnB 99 and UnB 110. In clone UnB 110, starch was observed in the cortical region, and medulla and gelatinous fibers were found in the pericycle and secondary xylem. Brown stem color was found to be associated with tolerance to drought.

ABSTRACT. Growth hormone (GH) is a part of the somatotropic axis that controls metabolism, growth, development and aging in a wide range of animals. Mutations that reduce GH signaling have been associated with extended life spans and increased longevity in ways similar to what is observed in dietary restriction (DR) models. However, the mechanism by which DR works is not well understood. Here, we show that DR works as a factor in the evolution of the genetic make-up of domestic cattle. In a series of 6864 bovines of seven Bos indicus and tropically adapted Bos taurus breeds, the frequency of a short, wild-type allele of the promoter region of the bovine GH gene, G1 allele, varied from 2.7 to 17.7%. The frequency of the long, domestic G2 allele increased from 88 to 95% along 20 calf crops of commercial Bos indicus cattle of the Nelore breed undergoing selection for increasing post-weaning weight gain with ad libitum nutrient intake. Under DR, however, the G1 allele sustained growth better than the G2 allele, as observed in a series of feeding tests. The G2 allele was even detrimental or abiotropic, as it caused rapid body decay under DR. We observed a reflection symmetry of GH allele substitution effects on body weight under different dietary schemes. The G2 allele is featured as the “demanding allele”, because it is optimally fitted to ad libitum nutrient intake. The G1 allele is featured as the “thrifty allele” because it is optimally fitted to DR. Our results show that dietary regimens need not extend lifespan or increase longevity in the sense of age-specific fitness. Instead, adaptation to any particular dietary regimen is just as much a consequence of selection as its cause; dietary regimens work as do any selection force, optimizing genotypic fitness to nutritional conditions.

ABSTRACT. The glutathione S-transferase (GST) family consists of phase II detoxification enzymes that catalyze the conjugation of toxic substances, such as chemotherapeutic agents, to glutathione. We examined whether GSTT1/GSTT1“null”, GSTM1/GSTM1“null” and GSTP1Ile105Ile/GSTP1Ile105Val polymorphisms are associated with different response rates to neoadjuvant chemotherapy in the treatment of stage II and III breast cancer. Forty Brazilian women with invasive ductal adenocarcinoma of the breast submitted to neoadjuvant chemotherapy, using 5-fluorouracil, epirubicin and cyclophosphamide, were genotyped for the GSTT1, GSTM1 and GSTP1 genes. Clinical response was assessed by RECIST criteria. Comparisons were made for the three genes alone and in pairs, as polymorphic and as wild-type combinations and polymorphic/wild-type combinations. We analyzed all possible combinations and their response rate. Patients with the GSTT1/GSTP1105Ile combination were found to have a significantly better response than GSTT1“null”/GSTP1105Val (P = 0.0209) and GSTT1/GSTM1 (P = 0.0376) combinations. Analysis of all possible combinations showed the GSTM1“null” polymorphic genotype to be present in four, and the wild-type GSTP1105Ile in six of the combinations associated with the largest number of responding patients. We found that patients with the GSTT1/GSTP1105Ile wild-type combination had a significantly higher response rate to chemotherapy than patients with the respective polymorphic GSTT1“null”/GSTP1105Val combination or patients with the wild-type GSTT1/GSTM1. The six gene combinations associated with the largest number of responding patients were found to contain the wild-type GSTP1105Ile and the polymorphic-type GSTM1“null”. These specific combinations were virtually absent in the combinations with few responding patients.

ABSTRACT. We evaluated the potential of the best linear unbiased predictor (BLUP) along with the relationship coefficient for predicting the performance of untested maize single-cross hybrids. Ninety S0:2 progenies arising from three single-cross hybrids were used. The 90 progenies were genotyped with 25 microsatellite markers, with nine markers linked to quantitative trait loci for grain yield. Based on genetic similarities, 17 partial inbred lines were selected and crossed in a partial diallel design. Similarity and relationship coefficients were used to construct the additive and dominance genetic matrices; along with BLUP, they provided predictions for untested single-crosses. Five degrees of imbalance were simulated (5, 10, 20, 30, and 40 hybrids). The correlation values between the predicted genotypic values and the observed phenotypic means varied from 0.55 to 0.70, depending on the degree of imbalance. A similar result was observed for the specific combining ability predictions; they varied from 0.61 to 0.70. It was also found that the relationship coefficient based on BLUP provided more accurate predictions than similarity-in-state predictions. We conclude that BLUP methodology is a viable alternative for the prediction of untested crosses in early progenies.

ABSTRACT. We looked for genotoxic effects in laboratory personnel routinely exposed to petroleum derivate compounds in an indoor environment. The exposed group of 21 workers from the Fuel Quality Control Laboratory of the Brazilian Petroleum Agency was matched with a group of 10 people from the staff of the Brazilian Ministry of Health. Chromosome aberrations in peripheral blood lymphocytes, micronuclei in exfoliated cells in the urine and hematological parameters were examined. There was a significantly increased level of chromosome aberrations and micronuclei in the exposed group compared with controls. A high correlation between chromosome aberrations and micronuclei was observed in the exposed group (Spearman rank test, r = 0.73, P = 0.0001). The hematological parameters in these exposed individuals did not differ from reference values.

ABSTRACT. The ae (amylose extender) recessive mutant alleles in maize are an important genetic resource for the development of high-amylose cultivars. On the basis of ae allele sequences (from the National Center for Biotechnology Information), the ae mutant alleles were cloned from high-amylose maize and the allelic Ae gene from common maize luyuan92 inbred lines. Five pairs of primers were designed to screen for a molecular marker of ae alleles, yielding a dominant molecular marker, ae474. We used 53 types of high-amylose maize and common maize inbred lines and their hybrid and backcross offspring for verification and analysis. The ae dominant molecular marker was effective in selecting for the ae alleles and for biological materials with a high-amylose genotype. Presence and absence of the marker in the offspring conformed to the expected Mendelian ratios. Using this marker, we were able to detect the ae alleles in a backcross and its second generation more efficiently (53.3 and 73.3%, respectively) than was possible without marker selection. These data indicate that the marker can be used as a tool to improve selection efficiency and accelerate the cultivation of new varieties of high-amylose maize.

ABSTRACT. Unlike other plants, bamboo (Bambusoideae) flowering is an elusive physiological phenomena, because it is unpredictable, long-periodic, gregarious, and uncontrollable; also, bamboo plants usually die after flowering. The flowering mechanism in Arabidopsis thaliana, a eudicot model species, is well established, but it remains unknown in bamboo species. We found 4470 and 3878 expressed sequence tags in the flower bud and vegetative shoot cDNA libraries, respectively, of the bamboo species, Bambusa oldhamii. Different genes were found expressed in bamboo flower buds compared to vegetative shoots, based on the Munich Information Center for Protein Sequences functional categorization; flowering-related genes were also identified in this species. We also identified Arabidopsis flowering-specific homologs that are involved in its photoperiod in this bamboo species, along with autonomous, vernalization and gibberellin-dependent pathways, indicating that bamboos may have a similar mechanism to control floral transition. Some bamboo expressed sequence tags shared high similarity with those of rice, but others did not match any known sequences. Our data lead us to conclude that bamboo may have its own unique flowering genes. This information can help us understand bamboo flowering and provides useful experimental methods to study the mechanisms involved.

ABSTRACT. We reviewed cytogenetic studies performed on 4216 patients who were referred to the Cytogenetics Unit at Dicle University Hospital, Diyarbair, Southeast Turkey, between 2000 and 2009. The cases were grouped according to the reason of referral for cytogenetic analysis. The frequencies of the different types of numerical and structural abnormalities were determined, and the relative frequency of cases with abnormal karyotypes was calculated in each group. The most common reason for requesting cytogenetic testing was referral for Down syndrome and for repeated abortions. The highest frequencies of abnormal karyotypes were found among cases that were referred due to suspicion of Down syndrome (84.8%). Among the chromosomal abnormalities, sexual chromosomal abnormalities were found in 239 cases (17.6%), and Klinefelter syndrome was the most frequent sex chromosomal abnormality. Autosomal abnormalities were found in 1119 cases (82.4%), and Down syndrome was the most frequent autosomal chromosomal abnormality. In conclusion, the high rate of chromosomal abnormalities (32.2%) found in this population demonstrates the importance of cytogenetic evaluation in patients who show clinical abnormalities. This is the first report on cytogenetic testing in the southeast region of Turkey. This type of study provides a basis for determining the risks of recurrence and for deciding on clinical treatment and genetic counseling.

ABSTRACT. The Mx (myxovirus resistance) gene codes for a protein with antiviral activity. Non-synonymous G/A polymorphism at position 2032 of chicken Mx cDNA results in a change at amino acid 631 of the Mx protein. This mutation has been shown to affect the antiviral activity of the Mx molecule, although recent studies have not confirmed this effect in response to some influenza strains. Nevertheless, the G/A polymorphism could be important for the chicken’s response to other viruses. A robust PCR-RFLP protocol for genotyping chicken Mx gene polymorphism associated with the S631N mutation was developed. The F primer anneals to the last intron of the Mx gene, and the R primer anneals to the last exon of the gene, with an expected PCR product of 299 bp. PCR products were digested with Hpy8I. This enzyme cuts the sequence 5’-GTN|NAC-3’, 2 bp downstream of the Mx polymorphism for the G allele, whereas the fragment containing the A allele is not cleaved. One hundred and twenty-seven chickens (commercial broilers, White Leghorn and New Hampshire) were genotyped using this protocol, and genotyping data were validated by sequencing. Full identity of results between the two genotyping methods was observed for all 127 samples, proving the reliability and robustness of this PCR-RFLP protocol.

ABSTRACT. CDKAL1 (cyckin-dependent kinase 5 regulatory subunit-associated protein 1-like 1) has been shown to be associated with type 2 diabetes in various ethnic groups; however, contradictory results have been reported. We performed a comprehensive meta-analysis of 21 studies for rs7756992, 17 studies for rs7754840 and 10 studies for rs10946398 variants of the CDKAL1 gene to evaluate the effect of CDKAL1 on genetic susceptibility for type 2 diabetes. We found a significant association of rs7756992, rs7754840 and rs10946398 in CDKAL1 with type 2 diabetes (odds ratio (OR) = 1.15, 95% confidence interval (CI) = 1.07-1.23, P < 0.0001; OR = 1.14, 95%CI = 1.06-1.24, P = 0.001, and OR = 1.12, 95%CI = 1.07-1.18, P < 0.0001, respectively). We conclude that there are significant associations between CDKAL1 polymorphisms and type 2 diabetes, but these associations vary in different ethnic populations.

ABSTRACT. We used the partially sequenced genomes of the turkey and chicken to find a large number of microsatellite markers. We then characterized 10 polymorphic microsatellite markers developed by cross-species amplification from economically and ecologically important birds to various European sub-species of the grey partridge. Even though we used cross-species amplification, a high degree of polymorphism was conserved in all microsatellite markers. Cross-species amplification from birds of economic and ecological interest, such as chicken and turkey, could be an attractive approach to develop microsatellite markers and to use these to manage wild and captive populations of other galliforms, such as the grey partridge.

ABSTRACT. The receptor for advanced glycation end products (RAGE or AGER) is a multiligand member of the immunoglobulin superfamily. RAGE is expressed in several tissues, including human myometrium, chorionic villi and placenta. Advanced glycation end products are the best studied ligands of RAGE; they have pro-inflammatory actions in human gestational tissues, increasing oxidative stress and the release of cytokines and prostaglandins. We investigated the association of RAGE gene promoter polymorphisms -429T>C (rs1800625) and -374T>A (rs1800624) with gestational diabetes. A sample of 750 unrelated European origin pregnant Brazilian women were classified as nondiabetic (control group, N = 600) or having gestational diabetes (N = 150) according to American Diabetes Association 2009 criteria. Genotyping was performed by PCR-RFLP. The frequencies of the rare alleles -429C (6.3 versus 9.1%) and -374A (26 versus 30%) were not significantly different between the gestational diabetes patients and healthy pregnant women. Also, the -429T>C and -374T>A polymorphisms were not associated with body mass index, lipid profile, fasting glycemia, HbA1C, or insulin requirement. We found that functional promoter polymorphisms of the RAGE gene were not associated with gestational diabetes or its complications in these Euro-Brazilian patients.

ABSTRACT. Amplified fragment length polymorphism (AFLP) analysis was carried out on representative individuals of wild Haliotis asinina using 64 primer combinations. Nine polymorphic AFLPs were cloned and sequenced. Sequence-specific primers were designed from six AFLP-derived fragments. Three sequence-characterized amplified region (SCAR) markers (HaSCAR₃₂₀, HaSCAR₂₉₅, HaSCAR₃₂₇) were selected for genotyping of 8-month-old domesticated stocks of H. asinina cultured separately at Sichang Marine Science Research and Training Station (N = 95) and at a hatchery in Trang province (N = 40) using single-strand conformational polymorphism analysis. Genotypes of wild abalone originating from Talibong Island (N = 25), Cambodia (N = 22), and the P₀ progeny established from Samet Island founders (N = 20) were also investigated. Significant genetic differentiation (P 0.0001 for the exact test and F_ST = 0.8759-0.8919, P 0.001) between abalone from the Gulf of Thailand (Cambodia and Samet Island - east) and the Andaman Sea (Talibong Island - west) were observed. This demonstrated the strong biogeographic structure of H. asinina in Thai waters. Non-overlapping composite genotypes for wild abalone from different coastal regions allow us to determine founder contributions in domesticated abalone stocks. Almost all Sichang Marine Science Research and Training Station and the Trang province hatchery stocks exhibited the east coast genotypes (97% of the 135 samples). We suggest that abalone from the east coast population have better survival rates under cultivated conditions than those from the west coast population.

ABSTRACT. Palicourea coriacea, popularly known as “douradinha”, is a medicinal plant from the Brazilian Cerrado region used in folk medicine to treat kidney and urethral stones and kidney inflammation. We evaluated the cytotoxic, genotoxic, and possible antigenotoxic activities of an aqueous extract of P. coriacea on somatic cells of Drosophila melanogaster, using the somatic mutation and recombination test. We used third-stage larvae of D. melanogaster from a standard cross and a high bioactivation cross and tested 10 different doses of P. coriacea aqueous extract (5, 15, 25, 35, 50, 65, 80, 95, 110, and 125 mg/mL). Doxorubicin (0.125 mg/mL) was used as a positive control and distilled water as a negative control. None of the doses was lethal to the larvae.There was no genotoxic effect at 5, 10, or 15 mg extract/mL. However, a significant decrease in the frequency of spots induced by doxorubicin was observed when administered with P. coriacea aqueous extract at these same doses. We conclude that P. coriacea aqueous extract is not cytotoxic or genotoxic at these doses, but it does protect against the genotoxic action of doxorubicin.

ABSTRACT. Soybean plants are promising bioreactors for the expression of biochemically complex proteins that cannot be produced in a safe and/or economically viable way in microorganisms, eukaryotic culture cells or secreted by transgenic animal glands. Soybeans present many desirable agronomic characteristics for high scale protein production, such as high productivity, short reproductive cycle, photoperiod sensitivity, and natural organs destined for protein accumulation in the seeds. The significant similarities between plant and human cells in terms of protein synthesis processes, folding, assembly, and post-translational processing are important for efficient accumulation of recombinant proteins. We obtained two transgenic lines using biolystics, incorporating the human proinsulin gene under control of the monocot tissue-specific promoter from sorghum γ-kafirin seed storage protein gene and the α-coixin cotyledonary vacuolar signal peptide from Coix lacryma-jobi (Poaceae). Transgenic plants expressed the proinsulin gene and accumulated the polypeptide in mature seeds. Protein targeting to cotyledonary protein storage vacuoles was successfully achieved and confirmed with immunocytochemistry assays. The combination of different regulatory sequences was apparently responsible for high stability in protein accumulation, since human proinsulin was detected after seven years under room temperature storage conditions.

ABSTRACT. Several lines of evidence suggest a molecular role of -1438A/G single nucleotide polymorphism in the 5-HTR2A gene promoter (rs6311) in regulating the expression of this gene, making rs6311 polymorphism a promising candidate for an association study. We looked for a possible association between rs6311 polymorphism and major depressive disorder (MDD) in a northeastern Thai population. We included 180 patients with MDD and 183 unrelated healthy controls in our study. Genotyping was performed using PCR-RFLP. We found no significant differences between the two groups with regard to both genotype distributions (X² = 1.32, d.f. = 2, P = 0.516) and allele frequencies (X² = 0.01, d.f. = 1, P = 0.913, odds ratio = 0.96, 95% confidence interval = 0.67-1.39). Therefore, this single nucleotide polymorphism appears not to be involved in the etiology of MDD.

ABSTRACT. We compared the expression of the ABCB1 gene in healthy male and female Thai subjects; this gene encodes the P-glycoprotein transporter in peripheral blood mononuclear cells (PBMCs). We also identified the most suitable housekeeping genes for normalization of ABCB1 expression levels in PBMCs. PBMCs from 30 females and 26 males were isolated. Total RNA was extracted, followed by reverse transcription (100 ng total RNA per sample). The internal normalization controls were actin-β, β-2M and GAPDH. Real-time quantitative PCR was then performed to determine the expression levels of the ABCB1 gene. The expression levels were found to be 1.5- to 2.5-fold higher in males, depending on the endogenous control used for normalization. Actin-β was the most stable control gene and could be used as a single endogenous control for normalization of ABCB1 expression levels in PBMCs. However, more than one endogenous control genes are recommended for normalization of gene expression. We conclude that the expression levels of ABCB1 in PBMCs is influenced by gender; this helps, in part, explain the gender difference in pharmacokinetics and pharmacodynamics of drugs that are P-glycoprotein substrates. ABCB1 gene expression profiles need to be carefully interpreted with regards to the endogenous control genes that are involved.

ABSTRACT. MicroRNAs (miRNAs) are non-coding small RNAs that regulate gene expression by translational repression or transcript degradation. A large number of miRNAs have been identified from model plant species; however, the character of conserved miRNAs is poorly understood. We studied 42 miRNA families that are conserved within the plant kingdom, using the miRBase database. Some conserved miRNA families were found to be preferentially expressed in dicots relative to monocots, especially miR403, miR472 and miR479. Using an improved homology search-based approach and the conserved miRNAs as the query set, 34 conserved miRNAs and the miR482 family were identified in wheat. Forty-six wheat mRNAs were predicted as their putative target genes. Most conserved wheat miRNAs were found to retain homologous target interactions and have analogous molecular functions. The miR172 displayed a wheat-specific function and was found to have an additional target interaction with succinyl-CoA ligase. We concluded that although miRNAs are conserved, the expression and function of some have drifted during long periods of plant evolution.

ABSTRACT. Association between neural tube defects (NTDs) and C677T polymorphism of the methylenetetrahydrofolate reductase (MTHFR) gene was suspected, because the MTHFR gene codes for a key enzyme in folate metabolism. Its deficiency usually leads to significant reductions in plasma concentrations of folate, vitamin B₁₂ and methionine, whereas homocysteine levels are increased. We examined folate, vitamin B₁₂ and homocysteine serum concentrations and polymorphism of the C677T MTHFR gene in Turkish children with neural tube defects. Thirty-three children with NTDs, 26 mothers and 48 healthy individuals were studied. C677T MTHFR polymorphism was determined by melting curve analyses (LightCycler®). The levels of folate, vitamin B₁₂ and homocysteine serum concentrations in NTDs were evaluated and compared, along with information concerning alleles of the MTHFR gene. C677T allele frequencies in NTD children and their mothers were similar to those found in controls. Serum folate and vitamin B₁₂ concentrations were significantly higher in NTD children than that of controls. Serum homocysteine concentrations were not significantly higher in NTD children and mothers. We concluded that C677T MTHFR gene polymorphism does not affect folic acid, vitamin B₁₂ and homocysteine metabolism in Turkish children with NTDs. C677T polymorphism of the MTHFR gene cannot be regarded as a major risk factor for NTDs in Turkish children.

ABSTRACT. Cabot’s tragopan, Tragopan caboti, is a globally threatened pheasant endemic to southeast China. The complete mitochondrial genome of Cabot’s tragopan was sequenced. The circular genome contains 16,727 bp, encoding a standard set of 13 protein-coding genes, two ribosomal RNA genes, and 22 transfer RNA genes, plus the putative control region, a structure very similar to that of other Galliformes. As found in other vertebrates, most of these genes code on the H-strand, except for the NADH dehydrogenase subunit 6 (nad6) and eight tRNA genes (Gln, Ala, Asn, Cys, Tyr, Ser(UCN), Pro, Glu). All protein-coding genes initiated with ATG, except for cox1, which began with GTG, and had a strong skew of C vs G (GC skew = -0.29 to -0.73). One extra ‘C’ nucleotide was found in the NADH dehydrogenase subunit 3 (nad3). All the tRNA gene sequences have the potential to fold into typical cloverleaf secondary structures. Conserved sequences in three domains were identified within the control region (D-loop). These results provide basic information for phylogenetic analyses among Galliform birds, and especially Tragopan species.

ABSTRACT. Thirteen isolates of Botryodiplodia theobromae collected from pear varieties grown in various regions of Punjab were studied for morphological, pathological and molecular characterization. The mycelial growth of B. theobromae isolates was classified as fluffy or depressed, uniform to irregular and cottony white turning to black. Colony growth rate varied from 19.1 to 24.9 mm per day. Pycnidia were produced either on the edge, centered or scattered on Petri dishes after 20 to 34 days of incubation. Pycnidia and pycnidiospores ranged in size from 118.0 to 240.0 μm and 14.5-35.5 x 6.5-14.5 μm, respectively. Lesion length produced by different isolates ranged from 1.9-7.2 x 0.8-3.3 cm with 49.4-90.9% infection. Using nine SSR and seven RAPD markers, amplified DNA bands ranged from 0.2 to 1.5 and 0.18 to 2.0 kb, respectively. Polymorphism information content values ranged from 0.44 to 0.71 and 0.63 to 0.93 for SSR and RAPD markers, respectively. A dendrogram based on molecular data, grouped the isolates into three major clusters with 65 to 79.5% genetic similarity; most of the isolates showed variety-specific grouping. The isolates prevalent on pear cultivars ‘Patharnakh’ and ‘Baggugosha’ in Ludhiana, Amritsar and Hoshiarpur districts were found to have a high degree of similarity; these isolates were also considerably distant from mango isolates and from other isolates on other pear cultivars. The isolates from cultivars Punjab Beauty, LeConte and Kieffer also had a high degree of similarity. Isolates from cultivar Smith were different from other pear isolates but showed more similarity with mango isolates.

ABSTRACT. We looked for a possible association between Klinefelter syndrome (KFS) and microdeletions in the Y chromosome in Turkish KFS patients. We examined the frequency of KFS in male patients with proven non-obstructive azoospermia and the types of Y chromosome microdeletions in these KFS patients. Fifty azoospermic patients and 50 fertile men were included in this study. KFS was found in 14 azoospermic patients. Y chromosome microdeletions were found in eight KFS patients. Azoospermia factor locus c (AZFc) was the most commonly deleted interval in KFS patients. All KFS patients had elevated plasma follicle-stimulating hormone and luteinizing hormone concentrations, but they had normal plasma testosterone concentrations. Testis biopsy of five samples with Y microdeletions revealed Sertoli cell-only syndrome. No Y microdeletions were found in the fertile group. We concluded that there could be an association between the AZFc region and KFS. Screening for this should be part of diagnostic work-up, particularly in those considering assisted reproduction.

ABSTRACT. The rock pigeon (Columba livia), or Rock dove, is a member of the bird family Columbidae. We mapped the complete mitochondrial genome of the Rock pigeon. The mitochondrial genome of this species is a circular molecule of 17,229 bp in length, encoding a standard set of 13 protein-coding genes, two ribosomal RNA genes, and 22 transfer RNA genes, plus a putative control region, demonstrating a structure very similar to that of other birds. As found in other vertebrates, most of these genes are coded on the H-strand, except for NADH dehydrogenase subunit 6 (nad6) and eight tRNA genes (Gln, Ala, Asn, Cys, Tyr, Ser(UCN), Pro, Glu). The AT skew and GC skew of the whole genome, protein-coding genes, tRNA, rRNA, and the control region were calculated for the complete mitochondrial genomes of 30 avian species, representing 29 orders. All protein-coding genes initiated with ATG, except for cox1 and nad5, which began with GTG. One extra nucleotide ‘C’ was present in NADH dehydrogenase subunit 3 (nad3). All tRNA gene sequences have the potential to fold into typical cloverleaf secondary structures. Within the control region, conserved sequences were identified in three domains. Although the conserved blocks, such as ETAS1, ETAS2, CSB1, CSB1-like, and boxes C, D, E, and F, are readily identifiable in the C. livia control region, the typical origin of H-strand replication (O_H), CSB2 and CSB3 could not be detected. These results provide basic information for phylogenetic analyses of birds, especially Columbiformes species.

ABSTRACT. The functions of distinct isoforms of solute carrier family 27 transporters (SLC27A1-6), acetyl-CoA carboxylase (ACACA, ACACB), stearoyl-CoA desaturase (SCD1-4), fatty acid desaturase (FADS1-3), LPIN (LPIN1-3), insulin-induced gene (INSIG1, 2), and peroxisome proliferator-activated receptor gamma coactivator1 (PPARGC1A, B) were studied in the mouse mammary gland from pregnancy to lactation. The relative mRNA abundance and percent change in real-time PCR were determined. mRNA expression of SLC27A3 and SLC27A4 was 37- and 1.4-fold more upregulated at 12 days of lactation, respectively (P < 0.01). Transcripts of SCD isoforms were the most abundant, accounting for 59% of all genes measured, and PPARGC1 isoforms were the least (0.06% of all genes measured). The mRNA abundance from ACC, FADS and LPIN accounted for 29, 9 and 2.6%, respectively. INSIG1 mRNA expression was 32-fold more upregulated (P < 0.05), while PPARGC1B was 0.18-fold downregulated at 18 days of lactation (P < 0.01). We concluded that mRNA abundance and expression of these isoforms are affected by the stage of lactation.

ABSTRACT. The biological cause of Pork Stress syndrome, which leads to PSE (pale, soft, exudative) meat, is excessive release of Ca2+ ions, which is promoted by a genetic mutation in the ryanodine receptors (RyR) located in the sarcoplasmic reticulum of the skeletal muscle cells. We examined the relationship between the formation of PSE meat under halothane treatment and heat stress exposure in chicken aRYR hot spot fragments. Four test groups were compared: 1) birds slaughtered without any treatment, i.e., the control group (C); 2) birds slaughtered immediately after halothane treatment (H); 3) birds slaughtered immediately after heat stress treatment (HS), and 4) birds exposed to halothane and to heat stress (H+HS), before slaughtering. Breast muscle mRNA was extracted, amplified by RT-PCR, and sequenced. PSE meat was evaluated using color determination (L* value). The most common alteration was deletion of a single nucleotide, which generated a premature stop codon, resulting in the production of truncated proteins. The highest incidence of nonsense transcripts came with exposure to halothane; 80% of these abnormal transcripts were detected in H and H+HS groups. As a consequence, the incidence of abnormal meat was highest in the H+HS group (66%). In HS, H, and C groups, PSE meat developed in 60, 50, and 33% of the samples, respectively. Thus, halothane apparently modulates αRYR gene expression in this region, and synergically with exposure to heat stress, causes Avian Stress syndrome, resulting in PSE meat in broiler chickens.

ABSTRACT. Infertility is a major reproductive health threat; the frequency of male infertility due to Y-chromosome microdeletions is 13-18% in the human population; these microdeletions involve recurrent loss of three non-overlapping regions designated as AZFa, AZFb and AZFc, associated with spermatogenic failure. Several contradictory reports have been published regarding deletion frequency based on sequence-tagged site markers and genotype-phenotype correlation. We examined the prevalence of Yq- deletion in 64 clinically diagnosed infertile male patients. We found a 3% frequency of microdeletion of the AZFc region; hormone profiles (FSH, LH and testosterone) showed significantly (P < 0.001) elevated levels compared to controls. No mutations were observed in the AZFa and AZFb regions, perhaps due to the selective use of sequence-tagged site markers.