Polymorphism in the third intron of the interferonγ gene is associated with susceptibility to multiple sclerosis.

The present study aims to examine the relationship between polymorphisms in the third intron of the IFN-γ gene and their influence on susceptibility to multiple sclerosis. A population-based case-control study was used for this purpose. Multiple sclerosis patients and healthy controls were interviewed. Genetic polymorphisms of IFN-γ intron III at the +2118 A/G and +3586 G/ACT sites were detected using polymerase chain reaction-restriction fragment length polymorphism. Genotypes and allele frequencies of IFN-γ intron III at the +2118 position were significantly different between multiple sclerosis patients and controls (P ≥ 0.05). However, no difference in allele frequencies was observed at the +3586 position between the two groups (P ≤ 0.05). Thus, polymorphisms at the +2118 A/G site in the IFN-γ intron III gene may be associated with susceptibility to multiple sclerosis.

Comparison of the antiplatelet effect of clopidogrel benzene sulfonate and clopidogrel hydrogen sulfate in stable coronary heart disease.

Clopidogrel hydrogen sulfate (CHS) is a thienopyridine, which can be used to prevent cardiovascular complications alone or in combination with acetyl salicylic acid as an important antiplatelet agent. Clopidogrel benzene sulfonate (CB) is a special clopidogrel salt that can be used as a conventional drug for antiplatelet effects, but the mechanism is still unknown. This study aimed to compare the antiplatelet effects of CHS and CB in stable coronary artery disease patients. Stable coronary artery disease patients (N = 119) were randomly divided into two groups receiving CHS (N = 67) or CB (N = 52). The patients were administered the drugs (600 mg dosage) and monitored for 12 to 14 h to detect antiplatelet effects. Antiplatelet response was evaluated by the P2Y12 response unit (PRU) and P2Y12 suppression percentage. In addition, all patients' CYP2C19*2, CYP2C19*3, and CYP3A5 polymorphisms were studied. Similar clinical manifestations were observed in the two groups. No obvious difference was detected in the platelet levels of patients given CHS or CB. The antiplatelet response (PRU and P2Y12 evaluation) of the patients using CHS and CB was not significantly different. In the two groups, the CYP2C19*2 polymorphic heterozygote number and antiplatelet response were similar. CB and CHS presented similar antiplatelet effects in stable coronary artery disease patients, and there was no difference in the CYP2C19*2 heterozygous polymorphism.

Assessment of correlation between pre-miRNA-1757 polymorphism and chicken performance traits.

Single nucleotide polymorphism in microRNAs (miRNA) may influence their target gene selection and regulation efficiency, leading to animal phenotypic variation. The aim of this study was to evaluate the possible effect of single nucleotide polymorphisms in the miRNA-1757 gene precursor region (pre-mir-1757) on economic-related traits in chicken. Genotyping was performed using Sequenom MassArray® iPLEX GOLD System. Association analysis was performed using SPSS19.0. The data showed that the G/C polymorphism was significantly correlated with semi-evisceration weight, evisceration weight, carcass weight, body weight at 10 weeks of age, shank length at 4 weeks of age, pectoral angle at 8 weeks of age, and body slanting length and pelvis breadth at 12 weeks of age (P < 0.05), and led to the alteration of the RNA secondary structure of pre-mir-1757. Our results provide useful information for further annotation studies of miRNA function.

Identification of an SCAR marker related to female phenotype in Idesia polycarpa Maxim.

Idesia polycarpa Maxim. is a dioecious species. Because of the lack of morphological and cytological methods available for identifying its sex during the long juvenile stage, the application of molecular markers in sex identification may facilitate sex determination in the seedling stage. The objective of this study was to use sequence-related amplified polymorphism to identify sex-linked markers in I. polycarpa and convert these markers into sequence-characterized amplified region markers, which are much easier to identify. A total of 342 primer combinations were screened and 2770 bands were examined. Only me14/em8 could amplify a specific fragment (210 base pairs) in all female but none in male plants. We analyzed this fragment using GenBank and found that the sequence similarity was 80% to the Populus trichocarpa clone POP006-H09 (sequence ID: gb|AC212923.1|) and that of the deduced amino acid sequence was 73% to the integrase of Mendicago truncatula (sequence ID: gb|ABD28291.1|) and 71% to the predicted retrotransposon integrase-like protein 1-like in Cicer arietinum (sequence ID: ref|XP 004515460.1|) (NCBI database through December 17, 2013). This fragment was converted into a stable and simple sequence-characterized amplified region marker approximately 200 base pairs in length. This marker can be utilized for early sexual identification in I. polycarpa, which will facilitate future breeding programs.

Cloning and characterization of a SnRK2 gene from Jatropha curcas L.

Although the SnRK2 class of Ser/Thr protein kinases is critical for signal transduction and abiotic stress resistance in plants, there have been no studies to examine SnRK2 in Jatropha curcas L. In the present study, JcSnRK2 was cloned from J. curcas using the rapid amplification of cDNA end technique and characterized. The JcSnRK2 genomic sequence is 2578 base pairs (bp), includes 10 exons and 9 introns, and the 1017-bp open reading frame encodes 338 amino acids. JcSnRK2 was transcribed in all examined tissues, with the highest transcription rate observed in the roots, followed by the leaves and stalks; the lowest rate was observed in flowers and seeds. JcSnRK2 expression increased following abscisic acid treatment, salinity, and drought stress. During a 48-h stress period, the expression of JcSnRK2 showed 2 peaks and periodic up- and downregulation. JcSnRK2 was rapidly activated within 1 h under salt and drought stress, but not under cold stress. Because of the gene sequence and expression similarity of JcSnRK2 to AtSnRK2.8, primarily in the roots, an eukaryotic expression vector containing the JcSnRK2 gene (pBI121-JcSnRK2) was constructed and introduced to the Arabidopsis AtSnRK2.8 mutant snf2.8. JcSnRK2-overexpressing plants exhibited higher salt and drought tolerance, further demonstrating the function of JcSnRK2 in the osmotic stress response. J. curcas is highly resistant to extreme salt and drought conditions and JcSnRK2 was found to be activated under these conditions. Thus, JcSnRK2 is potential candidate for improving crop tolerance to salt and drought stress.

Genetic diversity and relationships among Chinese Eucommia ulmoides cultivars revealed by sequence-related amplified polymorphism, amplified fragment length polymorphism, and inter-simple sequence repeat markers.

Sequence-related amplified polymorphism (SRAP), amplified fragment length polymorphism (AFLP), and inter-simple sequence repeat (ISSR) markers were used to estimate the genetic diversity and relationships among Eucommia ulmoides cultivars in China. A total of 240, 192, and 150 DNA fragments were detected by 10 SRAP primer combinations, 10 AFLP primer combinations, and 10 ISSR primers, among which 89.2, 65.1, and 88.0% of the fragments were polymorphic, respectively. Cluster analysis revealed that Qinzhong No. 3, Xiaoyeci, Qinzhong No. 1, and Qinzhong No. 2 formed independent clusters. The other 15 cultivars exhibited two clusters. The results of this study will help in the selection of parents for both genome mapping and crossbreeding purposes.

Relationship between progeny growth performance and molecular marker-based genetic distances in Eucommia ulmoides parental genotypes.

Progeny performances, variations and combining abilities for growth traits were evaluated in a factorial mating design of Eucommia ulmoides. Three marker systems, sequence-related amplified polymorphism, amplified fragment length polymorphism, and inter-simple sequence repeat, were used to determine genetic distances between parents. Correlations of genetic distances with progeny performances, within-family coefficients of variation and specific-combining abilities were established for height and basal diameter traits. Significant positive correlations were found between progeny performances of growth traits and genetic distances of parents based on sequence-related amplified polymorphism markers or a combination of all 3 marker systems. This revealed that crosses between genetically distant parents produced progenies with excellent growth performances. The lack of correlations between parental genetic distances and within-family coefficients of variation or specific-combining abilities suggested that these characteristics were unpredictable. The results of this study represent a potential criterion to predict progeny performances and choose parents in the breeding program.

Expression of ERCC1 and BRCA1 predict the clinical outcome of non-small cell lung cancer in patients receiving platinum-based chemotherapy.

We examined mRNA expression levels of ERCC1, BRCA1, RRM1, and human ß-tubulin-III (TUBB3) in non-small-cell lung carcinoma (NSCLC) patients and investigated the association between expression of these genes and the clinical outcome of NSCLC treatment. A total of 366 patients who underwent surgery for NSCLC were included in this study. All patients received third-generation platinum-based chemotherapy as first-line treatment. The relative cDNA quantification for ERCC1, RRM1, BRCA1, and TUBB3 was determined using a fluorescence-based, real-time detection method. We found that low expression of ERCC1 and BRCA1 was associated with a good response to platinum-based chemotherapy, with an odds ratio [95% confidence interval (CI)] of 2.09 (1.33-3.27) and 2.92 (1.85-4.62), respectively. Multivariate Cox regression analysis indicated that patients with low expression of ERCC1 and BRCA1 attained a longer overall survival time than those with high expression, with a hazard ratio (95%CI) of 0.42 (0.23-0.77) and 0.39 (0.21-0.71), respectively. However, RMM1 and TUBB2 expressions were not correlated with clinical outcome of NSCLC. In conclusion, we found that low expression of ERCC1 and BRCA1 can be useful for selecting NSCLC patients who would benefit from chemotherapy and warrants further investigation in prospective studies.

Genetic variations in the Wnt signaling pathway affect lung function in asthma patients.

Identifying genetic determinants for lung function is important in providing insight into the pathophysiology of asthma. The Wnt signal pathway plays a role in lung development and in asthma pathogenesis. However, whether genetic polymorphisms of Wnt signal pathway are associated with lung function in asthma patients remain unclear. We genotyped 2 single nucleotide polymorphisms (SNPs, rs2929973 and rs6581612) involved in the Wnt signal pathway in a cohort of 560 Chinese Han asthmatic children. Associations between each SNP and lung function, in a baseline exam, were tested using multiple linear regression models. We found that rs2929973 of the WISP1 gene was significantly associated with forced expiratory volume in 1 s (FEV1), with the G allele conferring significantly lower FEV1 values. However, the rs6581612 SNP of the WIF1 gene was not associated with differences in FEV1 values. We conclude that genetic variants in Wnt are associated with lung function and suggest that Wnt participates in inflammatory pathways that have an impact on the level of lung function.

Rapid determination of transgene copy number in tobacco by competitive PCR using a pair of SSR primers.

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.

[Evaluation of the nutritive value of soybean milk prepared with microwave-treated soybeans]. / Evaluación del valor nutricional de leches de soya preparadas a partir de granos de soya tratados con microondas.

Aqueous soybean extracts were prepared from beans, previously treated by microwaves to almost inactivate their lipoxygenase. These soy milks thus obtained were then nutritionally evaluated. All soy milks studied showed lower protein, fat, ash and total solids contents, as compared to a control milk prepared from soy beans not processed by microwaves. The milk obtained from soybeans with 8.7% initial moisture, treated by microwaves for 240 seconds, had the best total chemical score and the highest apparent methionine availability, as well as PER. The complete inactivation of the trypsin inhibitor activity was achieved with the milk prepared from soybeans with 56.8% initial moisture, subjected to microwave treatment for 180 seconds. On the other hand, the milk obtained from soybeans with 38.8% initial moisture, processed by microwaves for 180 seconds, resulted to have the highest, in vitro, protein digestibility.

Evaluación del valor nutricional de leches de soya preparadas a partir de granos de soya tratados con microondas / Evaluation of the nutritive value of soybean milk prepared with microwave-treated soybeans

Se prepararon extractos acuosos de soya a partir de granos previamente sometidos a microondas, para la inactivación casi completa de la lipoxigenasa. Las leches de soya así obtenidas se evaluaron nutricionalmente. Todas las leches sometidas a estudio presentaron menor contenido de proteína, lípidos, ceniza y sólidos totales, en comparación con el alimento control, preparado a partir de granos no tratados por microondas. La leche obtenida de granos de soya con 8.7% de humedad inicial, irradiados con microondas durante 240 segundos, acusó el mejor puntaje químico de aminoácidos esenciales y la mayor disponibilidad aparente de metionina y PER. La inactivación completa de la actividad inhibidora de la tripsina se obtuvo en la leche preparada a partir de granos de soya con 56.8% de humedad, tratados por microondas durante 180 segundos. Por otro lado, la leche obtenida de granos de soya con 38.8% de humedad, irradiados con microondas durante 180 segundos, resultó ser la de mayor digestibilidad de la proteína in vitro