Nucleotide Polymorphism Sites Research Articles

Genomic Selection for F1 Hybrid Breeding in Strawberry (Fragaria × ananassa)

Cultivated strawberry is the most widely consumed fruit crop in the world, and therefore, many breeding programs are underway to improve its agronomic traits such as fruit quality. Strawberry cultivars were vegetatively propagated through runners and carried a high risk of infection with viruses and insects. To solve this problem, the development of F1 hybrid seeds has been proposed as an alternative breeding strategy in strawberry. In this study, we conducted a potential assessment of genomic selection (GS) in strawberry F1 hybrid breeding. A total of 105 inbred lines were developed as candidate parents of strawberry F1 hybrids. In addition, 275 parental combinations were randomly selected from the 105 inbred lines and crossed to develop test F1 hybrids for GS model training. These populations were phenotyped for petiole length, leaf area, Brix, fruit hardness, and pericarp color. Whole-genome shotgun sequencing of the 105 inbred lines detected 20,811 single nucleotide polymorphism sites that were provided for subsequent GS analyses. In a GS model construction, inclusion of dominant effects showed a slight advantage in GS accuracy. In the across population prediction analysis, GS models using the inbred lines showed predictability for the test F1 hybrids and vice versa, except for Brix. Finally, the GS models were used for phenotype prediction of 5,460 possible F1 hybrids from 105 inbred lines to select F1 hybrids with high fruit hardness or high pericarp color. These F1 hybrids were developed and phenotyped to evaluate the efficacy of the GS. As expected, F1 hybrids that were predicted to have high fruit hardness or high pericarp color expressed higher observed phenotypic values than the F1 hybrids that were selected for other objectives. Through the analyses in this study, we demonstrated that GS can be applied for strawberry F1 hybrid breeding.

The combination of methylenehydrofolate reductase C677T polymorphism screening and gastrointestinal tumor markers detection may be an early screening method for gastrointestinal cancer related to helicobacter pylori infection

Methyltetrahydrofolate reductase (MTHFR) is a key enzyme in folate metabolism, and its single nucleotide polymorphism (SNP) site C677T may be associated with gastrointestinal cancer. However, the relationship between MTHFR C677T polymorphism and gastrointestinal tumor markers carcinoma embryonic antigen (CEA), carbohydrate antigen 199 (CA199) and carbohydrate antigen 724 (CA724) in Helicobacter pylori (H. pylori) infection is not specified. This study aims to identify the association between MTHFR C677T polymorphism and gastrointestinal tumor markers (CEA, CA199 and CA724) in H. pylori infection. The relationship between MTHFR C677T polymorphism and gastrointestinal tumor markers in 58 patients with H. pylori infection and 94 non-infected patients was studied. We found that TT genotype was a susceptibility factor of H. pylori infection, which was also associated with increased CEA and CA724 levels. Moreover, there was a negative additive interaction between MTHFR gene C677T polymorphism and CEA levels in H.pylori infection. Meanwhile, there were significant differences in CEA levels between MTHFR C677T polymorphism and H.pylori infection. The presence of T allele led to a decrease in CEA levels when 13C urea breath test (13C-UBT) was positive, while the presence of T allele led to an increase in CEA levels when 13C-UBT was negative. Therefore, we suggest that healthy people should take MTHFR C677T polymorphism screening, combined with 13C-UBT and gastrointestinal tumor markers detection, which can screen out the susceptible population of H. pylori, and help to detect gastrointestinal cancer in the early stage.

Association between methylation of BIN1 promoter in peripheral blood and preclinical Alzheimer\u2019s disease

The bridging integrator 1 (BIN1) gene is the second most important susceptibility gene for late-onset Alzheimer’s disease (LOAD) after apolipoprotein E (APOE) gene. To explore whether the BIN1 methylation in peripheral blood changed in the early stage of LOAD, we included 814 participants (484 cognitively normal participants [CN] and 330 participants with subjective cognitive decline [SCD]) from the Chinese Alzheimer’s Biomarker and LifestylE (CABLE) database. Then we tested associations of methylation of BIN1 promoter in peripheral blood with the susceptibility for preclinical AD or early changes of cerebrospinal fluid (CSF) AD-related biomarkers. Results showed that SCD participants with significant AD biological characteristics had lower methylation levels of BIN1 promoter, even after correcting for covariates. Hypomethylation of BIN1 promoter were associated with decreased CSF Aβ42 (p = 0.0008), as well as increased p-tau/Aβ42 (p = 0.0001) and t-tau/Aβ42 (p < 0.0001) in total participants. Subgroup analysis showed that the above associations only remained in the SCD subgroup. In addition, hypomethylation of BIN1 promoter was also accompanied by increased CSF p-tau (p = 0.0028) and t-tau (p = 0.0130) in the SCD subgroup, which was independent of CSF Aβ42. Finally, above associations were still significant after correcting single nucleotide polymorphic sites (SNPs) and interaction of APOE ɛ4 status. Our study is the first to find a robust association between hypomethylation of BIN1 promoter in peripheral blood and preclinical AD. This provides new evidence for the involvement of BIN1 in AD, and may contribute to the discovery of new therapeutic targets for AD.

The Correlation Between FGB Promoter Polymorphism and Clotting Function in Patients With Idiopathic Lower Extremity Deep Venous Thrombosis.

To explore the possible single nucleotide polymorphisms (SNPs) sites in the promoter region of fibrinogen B β (FGB), and construct logistic regression model and haplotype model, so as to reveal the influence of FGB promoter SNPs on susceptibility, hemodynamics and coagulation function of lower extremity deep venous thrombosis (LEDVT) in the genetic background. LEDVT patients (120) and healthy people (120) were taken as case and control objects, respectively. SNPs and their genotypes of FGB promoter were detected by promoter sequencing and PCR-RFLP. The parameters of coagulation system were evaluated. There were 6 SNPs in FGB promoter, which were β-148C/T, β-249C/T, β-455G/A, β-854G/A, β-993C/T and β-1420G/A. The genotype and allele frequency of β-1420 G/A, β-455G/A, β-249c/T and β-148C/T were significantly different between the LEDVT group and the control group, but not β-993C/T and β-854G/A. In addition, we found that the higher the content of Fibrinogen (FG), the higher the risk of LEDVT. The risk of LEDVT increased by 4.579 times for every unit increase of fibrinogen. We also found that FG, PT and APTT in LEDVT group were higher than those in control group, while TT was lower than those in control group; Furthermore, there was no significant difference in all coagulation indexes among 6 SNP genotypes in LEDVT group, while a significant difference was found between the 2 genotypes of β-993C/T in the control group. β-993C/T may indirectly affect the susceptibility of LEDVT by improving the basic level of plasma FG.

The expression of NLK is functionally associated with colorectal cancers (CRC).

The regulatory mechanism of NLK in the carcinomagenesis and progression of colorectal cancer (CRC) remains unclear. Here, we identified a single nucleotide polymorphism (SNP) site of NLK (rs2125846) as a new susceptibility locus for CRC risk located within an intron of the human NLK gene in a Chinese population. NLK downregulation led to a decrease in the ability of proliferation and migration of RKO cells in vitro. The proportion of RKO apoptotic cells increased by interfering with the endogenous expression of NLK. We speculate that LncRNA XIST may upregulate NLK expression by downregulating miR-92b-3p, thereby promote the development of CRC. These results provide important information for the identification of novel potential targets for the prevention or treatment of CRC.

FTO - A Common Genetic Basis for Obesity and Cancer.

In recent years, the prevalence of obesity and cancer have been rising. Since this poses a serious threat to human health, the relationship between the two has attracted much attention. This study examined whether fat mass and obesity-associated (FTO) genes are linked, taking into account a Genome-wide Association Study (GWAS) that revealed multiple single nucleotide polymorphism sites (SNPs) of the FTO gene, indicating an association between obesity and cancer in different populations. FTO proteins have been proved to participate in adipogenesis and tumorigenesis with post-transcriptional regulation of downstream molecular expression or through the target of the mammalian target protein rapamycin (mTOR). FTO inhibitors have also been found to share anti-obesity and anti-cancer effects in vivo. In this review, we comprehensively discuss the correlation between obesity and cancer by measuring FTO gene polymorphism, as well as the molecular mechanism involved in these diseases, emphasizing FTO as the common genetic basis of obesity and cancer.

The Pursuit of COVID-19 Biomarkers: Putting the Spotlight on ACE2 and TMPRSS2 Regulatory Sequences.

Diverse populations worldwide are differentially affected by coronavirus disease 2019 (COVID-19). While socioeconomic background has been studied extensively, little is known about the genetic variation underlying this phenomenon. This study is aimed at examining the genetic basis behind the great discrepancies among diverse ethnic groups in terms of COVID-19 susceptibility for viral infection, disease prognosis, and mortality. To this end, in silico analysis of single-nucleotide polymorphisms (SNPs) within regulatory sequences of the human angiotensin-converting enzyme 2 (ACE2) and transmembrane protease serine 2 (TMPRSS2)—the virus's gateway to host cells—and their plausible implications on expression levels was conducted. We provide indication that the variation in the human ACE2 and TMPRSS2 regulatory sequences is likely to be involved in and contribute to this phenomenon. SNPs that are abundant in the more susceptible populations introduce binding sites (BSs) for transcription factors or they may invalidate BSs for transcription repressor—both may enhance target gene (ACE2 or TMPRSS2) expression in the relevant target tissues. SNPs that are abundant in the more resistant populations may invalidate BSs for a transcriptional repressor or they may introduce BSs for a transcriptional repressor or initiator of mRNA degradation, which may reduce target gene expression levels. This aspect, when added to the socioeconomic factors, can be a cause for the divergent prevalence of the disease and the different mortality rates within diverse populations. This demonstration may call for a shift in the paradigm of searching for COVID-19 biomarkers, such that SNPs within regulatory sequences should be of high importance.

Tracking the history of grapevine cultivation in Georgia by combining geometric morphometrics and ancient DNA

The Near East and the Caucasus are commonly regarded as the original domestication centres of Vitis vinifera (grapevine), and the region continues to be home to a high diversity of wild and cultivated grapevines, particularly within Georgia. The earliest chemical evidence for wine making was recorded in Georgian Neolithic sites (6000–5800 bc) and grape pips, possibly of the domesticated morphotype, have been reported from several sites of about the same period. We performed geometric morphometric and palaeogenomic investigations of grape pip samples in order to identify the appearance of domesticated grapevine and explore the changes in cultivated diversity in relation to modern varieties. We systematically investigated charred and uncharred grape pip samples from Georgian archaeological sites. Their chronology was thoroughly assessed by direct radiocarbon dating. More than 500 grape pips from 14 sites from the Middle Bronze Age to modern times were selected for geometric morphometric studies. The shapes of the ancient pips were compared to hundreds of modern wild individuals and cultivated varieties. Degraded DNA was isolated from three pips from two sites, converted to Illumina libraries, sequenced at approximately 10,000 single nucleotide polymorphism (SNP) sites, and compared to a large public database of grapevine diversity. The most ancient pip dates from the Middle Bronze Age (1900–1500 cal bc) and the domesticated morphotype is identified from ca. 1000 bc onwards. A great diversity of domesticated shapes was regularly seen in the samples. Most are close to modern cultivars from the Caucasian, southwest Asian and Balkan areas, which suggests that the modern local vine diversity is deeply rooted in early viticulture. DNA was successfully recovered from historic pips and genome-wide analyses found close parental relationships to modern Georgian cultivars.

Novel SNP markers on ginsenosides biosynthesis functional gene for authentication of ginseng herbs and commercial products

Panax ginseng and Panax quinquefolius have similar bioactive components and morphological characteristics, but they are known to have different medicinal values, high-sensitive and accurate method is expected to identify the sources of ginseng products and evaluate the quality, but with a huge challenge. Our established UHPLC-TOF/MS method coupled with orthogonal partial least squares discriminant analysis (OPLS-DA) model based on 18 ginsenosides was applied to discriminate the sources of raw medicinal materials in ginseng products, and nested PCR strategy was used to discover 6 novel single nucleotide polymorphism (SNP) sites in functional dammarenediol synthase (DS) gene for genetic authentication of P. ginseng and P. quinquefolius for the first time. OPLS-DA model could identify the sources of raw ginseng materials are real or not. SNP markers were applied to identify ginseng fresh samples as well as commercial products, and proved to be successful. This established molecular method can tell exact source information of adulterants, and it was highly sensitive and specific even when total DNA amount was only 0.1 ng and the adulteration was as low as 1%. Therefore, this study made an attempt at the exploration of new type SNP marker for variety authentication and function regulation at the same time, and the combination of chemical and molecular discrimination methods provided the comprehensive evaluation and authentication for the sources of ginseng herbs and products.

Analysis of Correlation between Gene Polymorphisms of CRHBP in Yunnan Han Population and Schizophrenia and Aggressive Behavior

Objective To explore the correlation between 4 tag single nucleotide polymorphisms （Tag SNP） sites （rs7721799, rs32897, rs7718461, rs10062367） of corticotropin releasing hormone binding protein （CRHBP） and schizophrenia and aggressive behavior in the Yunnan Han population. Methods Case-control correlation analysis was used to establish a complex amplification system. Improved multiplex ligase detection reaction （iMLDR） technology was used to detect the genotypes of 4 SNP sites of CRHBP gene of 163 Han schizophrenic patients （including 81 patients with aggressive behavior, 82 patients without aggressive behavior） and 345 healthy Han individuals, which were analyzed statistically by SPSS 19.0, Haploview 4.2 and PHASE 2.1 software. Results There was no correlation between the 3 SNP sites of CRHBP gene and the onset of schizophrenia except for the rs7718461 site （P>0.05）. The relative risk of aggressive behavior of patients carrying GG or GA genotype at rs7718461 site were 4.903 times higher than those carrying AA genotype （P<0.05）. Conclusion The CRHBP gene may not be associated with the occurrence of schizophrenia in Yunnan Han population, but AA genotype of rs7718461 may reduce the risk of aggressive behavior in schizophrenia patients.

A new naturalized plant in Korea: Carex molestiformis Reznicek and Rothrock (Cyperaceae)

국내 ë¯¸ê¸°ë¡ì¢ ê°€ëŠ”íƒ€ëž˜ì‚¬ì´ˆ(Carex molestiformis Reznik and Rothrock)를 강원도 평창군 ë²½íŒŒë ¹ 일대 에서 처음으로 발견하였다. 본 분류군의 원산지는 북아메리카 미국의 남부지역(Missouri, Oklahoma, Arkansas등 11개 주)으로, 자생지에서는 주로 개활지나 강둑, 범람원 등에서 무리지어 밀생한다. íƒ€ëž˜ì‚¬ì´ˆì ˆ (sect. Ovales)에 속하며, ìžì› ì„± 소수를 갖는 ë‹¤ë „ìƒ 초본으로, 한 화서에 달리는 소수의 수가 2–4개로 ì ê³ , 과낭의 폭이 2.6–3.4 mm로 넓으며, 과낭 배면의 맥이 6–9개로 많아 국내에 자생하는 ê·¼ì—°ì¢ ì¸ 타래사초(C. maackii Maxim.) 및 ê·€í™”ì¢ ì¸ 한석사초(C. scoparia)와 ëª í™•ížˆ 구별된다. êµ­ëª ì€ 본 분류군이 í˜•íƒœì ìœ¼ë¡œ 타 래사초와 ìœ ì‚¬í•˜ë©°, 식물체가 ì „ì²´ì ìœ¼ë¡œ 가늘어 ‘가는타래사초’로 ì‹ ì¹­í•˜ì˜€ê³ , 분류군에 대한 기재문, 화상 자료 및 도해 ê·¸ë¦¬ê³ ìœ ì‚¬ì¢ ì— 대한 검색표를 ì œì‹œí•˜ì˜€ë‹¤. 또한, 가는타래사초와 ê·¼ì—°ì¢ ì˜ ìœ ì „ì  차이를 확 ì¸í•˜ê³ ìž 3개(chloroplast DNA matK, ndhF, nuclear ribosomal DNA internal transcribed spacer) DNA barcode 지역의 부분 염기서열을 비교하여, 총 8개의 가는타래사초의 ì¢ íŠ¹ì´ì  단일염기다형성(single nucleotide polymorphisms)을 확인하였다. 주요어: ë¯¸ê¸°ë¡ì¢ , 가는타래사초, íƒ€ëž˜ì‚¬ì´ˆì ˆ, 사초과

Point-of-care DNA testing by automatically and sequentially performing extraction, amplification and identification in a closed-type cassette

Nucleic acid detection is important for clinical diagnostics; however, it is challenging to perform genetic testing at the point-of-care due to the tedious steps involved in DNA extraction and the risk of cross-contamination from amplicons. To achieve a fully-automated and contamination-free nucleic acid detection, we propose a closed-type cassette system which enables the following steps to be operated automatically and sequentially: sample preparation based on magnetic beads, target amplification using multiplex polymerase chain reaction, and colorimetric detection of amplicons using a serial invasive reaction coupled with the aggregation of gold nanoparticle probes. The cassette was designed to be round and closed, and 10 targets in a sample could be simultaneously detected by the naked eye or using a spectrophotometer in the system. In addition, a cassette-driven device was fabricated to transfer reagents between wells, to control the temperature of each reaction, and to sense the colour in the detection wells. The cassette system was sensitive enough to detect 10 genotypes at 5 single nucleotide polymorphism sites related to the anticoagulant’s usage, by using a 0.5 μL blood sample. The accuracy of the system was evaluated by detecting 12 whole blood samples, and the results obtained were consistent with those obtained using pyrosequencing. The cassette is airtight and the whole system is fully automatic; the only manual operation is the addition of the sample to the cassette, performing point-of-care genetic testing in a sample-in/answer-out way.

Mining Listeria monocytogenes single nucleotide polymorphism sites to identify the major serotypes using allele-specific multiplex PCR

Worldwide, Listeria monocytogenes is a common foodborne pathogen and serotyping is necessary for traceability and surveillance. Due to high accuracy and speed, PCR is commonly used for serotyping by targeting specific genes, such as lmo1118 and ORF2110 for 1/2c-3c and 4b-4d-4e, respectively. Single nucleotide polymorphisms (SNPs) are single base alterations at specific loci which are associated with various phenomena. In this study, a method was explored to mine specific SNPs from 253 L. monocytogenes genomes with known serotypes by writing Python programming language script. After blasting all the CDS, seventeen candidate genes with specific SNPs were obtained and these genes contained multiple types of SNPs, including lineages I, II, III, 1/2a-3a, 1/2b-3b-7, and 1/2c-3c specific SNPs. The sensitivity and specificity of these candidate SNP sites are higher than 85% in the genomes examined. Combining lineage I-specific, lineage II-specific, 1/2b-3b-7-specific, and 1/2c-3c-specific SNP sites together, using allele-specific multiplex PCR, we could serotype major L. monocytogenes serotypes. This method was used for 60 L. monocytogenes strains isolated from various food samples and the serotyping results were 100% identical to those of the currently accepted method with the reduced primers from ten to eight. Our results indicate that allele-specific multiplex PCR after mining specific SNPs from common genome database can be used for bacterial typing.

The Husavirus Posa-Like Viruses in China, and a New Group of Picornavirales.

Novel posa-like viral genomes were first identified in swine fecal samples using metagenomics and were designated as unclassified viruses in the order Picornavirales. In the present study, nine husavirus strains were identified in China. Their genomes share 94.1–99.9% similarity, and alignment of these nine husavirus strains identified 697 nucleotide polymorphism sites across their full-length genomes. These nine strains were directly clustered with the Husavirus 1 lineage, and their genomic arrangement showed similar characteristics. These posa-like viruses have undergone a complex evolutionary process, and have a wide geographic distribution, complex host spectrum, deep phylogenetic divergence, and diverse genomic organizations. The clade of posa-like viruses forms a single group, which is evolutionarily distinct from other known families and could represent a distinct family within the Picornavirales. The genomic arrangement of Picornavirales and the new posa-like viruses are different, whereas the posa-like viruses have genomic modules similar to the families Dicistroviridae and Marnaviridae. The present study provides valuable genetic evidence of husaviruses in China, and clarifies the phylogenetic dynamics and the evolutionary characteristics of Picornavirales.

The impact of EGFR gene polymorphisms on the response and toxicity derived from neoadjuvant chemotherapy for breast cancer.

Neoadjuvant chemotherapy is usually used for treating locally advanced breast cancer. However, not all patients achieve pathologic complete response (pCR). In this study, we selected two epidermal growth factor receptor (EGFR) single nucleotide polymorphism (SNP) sites, rs1468727 and rs845552, to investigate the association between the genotypes and the response and toxicity derived from neoadjuvant chemotherapy for breast cancer. All participants took part in clinical trial SHPD001 and SHPD002. For univariate analyses, the association between SNP and pCR or toxicity was analyzed by Chi-square or Fisher's exact test. For multivariate analyses, logistic regression was used instead. In all, one hundred and eighteen patients were enrolled. We found that the frequency of AA genotype in rs845552 was higher than that of other genotypes in HER2-positive breast cancer (AA vs. AG, P=0.039; AA vs. GG, P=0.005; AA vs. AG+GG, P=0.009). Multivariate logistic regression analyses showed that pCR was more difficult to be achieved in patients with a CT genotype in rs1468727 compared to those with a CC+TT genotype (OR =0.288, 95% CI: 0.109-0.762, P=0.012) or a CC genotype (OR =0.254, 95% CI: 0.076-0.849, P=0.026). Moreover, we demonstrated that both rs1468727 and rs845552 were associated with toxicity that results in complications such as increased total bilirubin, skin rash, peripheral neuropathy, and alopecia (P<0.05). Our study reported for the first time, that in treating breast cancer with neoadjuvant chemotherapy, EGFR SNP rs1468727 is associated with treatment response, and that both rs1468727 and rs845552 are related to treatment-derived toxicity. In addition, we also found that rs845552 may be related to the status of HER2 in breast cancer.

Evolutionary directions of single nucleotide substitutions and structural mutations in the chloroplast genomes of the family Calycanthaceae

BackgroundChloroplast genome sequence data is very useful in studying/addressing the phylogeny of plants at various taxonomic ranks. However, there are no empirical observations on the patterns, directions, and mutation rates, which are the key topics in chloroplast genome evolution. In this study, we used Calycanthaceae as a model to investigate the evolutionary patterns, directions and rates of both nucleotide substitutions and structural mutations at different taxonomic ranks.ResultsThere were 2861 polymorphic nucleotide sites on the five chloroplast genomes, and 98% of polymorphic sites were biallelic. There was a single-nucleotide substitution bias in chloroplast genomes. A → T or T → A (2.84%) and G → C or C → G (3.65%) were found to occur significantly less frequently than the other four transversion mutation types. Synonymous mutations kept balanced pace with nonsynonymous mutations, whereas biased directions appeared between transition and transversion mutations and among transversion mutations. Of the structural mutations, indels and repeats had obvious directions, but microsatellites and inversions were non-directional. Structural mutations increased the single nucleotide mutations rates. The mutation rates per site per year were estimated to be 0.14–0.34 × 10− 9 for nucleotide substitution at different taxonomic ranks, 0.64 × 10− 11 for indels and 1.0 × 10− 11 for repeats.ConclusionsOur direct counts of chloroplast genome evolution events provide raw data for correctly modeling the evolution of sequence data for phylogenetic inferences.

Deep learning for population size history inference: Design, comparison and combination with approximate Bayesian computation.

For the past decades, simulation-based likelihood-free inference methods have enabled researchers to address numerous population genetics problems. As the richness and amount of simulated and real genetic data keep increasing, the field has a strong opportunity to tackle tasks that current methods hardly solve. However, high data dimensionality forces most methods to summarize large genomic data sets into a relatively small number of handcrafted features (summary statistics). Here, we propose an alternative to summary statistics, based on the automatic extraction of relevant information using deep learning techniques. Specifically, we design artificial neural networks (ANNs) that take as input single nucleotide polymorphic sites (SNPs) found in individuals sampled from a single population and infer the past effective population size history. First, we provide guidelines to construct artificial neural networks that comply with the intrinsic properties of SNP data such as invariance to permutation of haplotypes, long scale interactions between SNPs and variable genomic length. Thanks to a Bayesian hyperparameter optimization procedure, we evaluate the performance of multiple networks and compare them to well-established methods like Approximate Bayesian Computation (ABC). Even without the expert knowledge of summary statistics, our approach compares fairly well to an ABC approach based on handcrafted features. Furthermore, we show that combining deep learning and ABC can improve performance while taking advantage of both frameworks. Finally, we apply our approach to reconstruct the effective population size history of cattle breed populations.

Sequence analysis based on ITS1 region of nuclear ribosomal DNA of Amomum villosum and ten species of Alpinia

Ripe fruits of Amomum villosum Lour. are important traditional Chinese medicines and food spices. Due to similar morphological and anatomical characteristics, fruits from some Alpinia species are used clinically as substitutes or adulterants of Fructus Amomi. In this study, we determined the nrDNA ITS1 (internal transcribed space 1) sequences from Amomum villosum and ten Alpinia species, which are adulterants of Fructus Amomi. The ITS1 sequences of these species were 175-179 bp in length, and the G+C contents were 53.37-56.74%. Sequence alignment revealed that there were 55 polymorphic nucleotide sites among the tested species. The base substitution were 0.57-9.60% within the ten Aplinia species, but 17.61-21.59% between Amomum villosum and the Alpinia species. These molecular data indicated that ITS1 sequences can be used to differentiate Fructus Amomi and its substitutes or adulterants from various Alpinia species.

Studying of cyp2c19*2, *3 and *17 polymorphism in Vietnamese patients with coronary artery disease

Coronary artery disease is the most common type of cardiovascular disease, due to the accumulation of atherosclerotic plaque inside the arterial wall which leads to block blood supply to the heart muscle. A number of clinical trials have demonstrated that Clopidogrel is able to inhibit platelet aggregation in patients with acute coronary syndromes, reduce mortality and cardiovascular events. However, the antiplatelet effectiveness of Clopidogrel significantly depends on CYP2C19 genotypes. Therefore, the aim of this study was to identify the CYP2C19*2, *3 and allele frequencies in Vietnamese coronary artery patients by using PCR-RFLP method. Total genomic DNA were extracted from peripheral blood of 96 patients diagnosed with coronary artery disease. Thereafter, single nucleotide polymorphism sites in the CYP2C19 gene were identified by PCR with specific primers. The amplified products were then digested by restriction enzymes SmaI, BamHI, and MnlI, respectively. The results showed that the proportion of heterozygous individuals for CYP2C19*2 (c. 681G&gt;A, rs4244285), CYP2C19*3 (c. 636G&gt;A, rs4986893), and CYP2C19*17 (g. -3402C&gt;T, rs11188072) accounted for 39.58%, 6.25%, and 2.08%, respectively. Among 96 subjects, 41.67% of patients were predicted for intermediate metabolic phenotype CYP2C19*1/*2 (37.50%) and CYP2C19*1/*3 (4.17%). Approximately 10.42% of total patients represented poor metabolizers in which 8.34% had two copies of the same allele *2/*2 and 2.08% had *2/*3 genotype. Particularly, two individuals (2.08%) detected with CYP2C19*1/*17 genotype were able to increase CYP2C19 activity (ultrarapid metabolizers). The results of this study generate a foundation for introducing individualized antiplatelet therapy in Vietnam based on genetic testing.

Practical Application of Miyazaki Jitokko Chickens Selected for a Superior Allele at a Single Nucleotide Polymorphism Site in the Cholecystokinin Type A Receptor Gene.

This study aimed to examine 1) whether selection for a superior allele at a single nucleotide polymorphism site (SNP; AB604331, g.420 C>A) of the chicken cholecystokinin type A receptor (CCKAR) gene in Miyazaki Jitokko chickens is detectable in commercial poultry farms, and 2) whether the reproductive traits of the Kyushu Rhode hens, as a maternal stock line of the Miyazaki Jitokko chickens, are affected by SNP selection. Conventional and A-allele fixed (improved) Miyazaki Jitokko chicks were hatched on the same day and raised in a battery cage until 7 days of age. The chicks were then deposited at two commercial poultry farms and reared until slaughter at 126 and 163 days for cockerels and pullets, respectively. Body weight on the day of hatching (day 0), at 5 days of age, and at slaughter were measured. The differences in the body weights of the farm and test groups at slaughter were analyzed using the generalized linear model. A-allele fixation increased the body weight at slaughter by approximately +123.5 g and +131.9 g in cockerels and pullets, respectively. No significant differences between the conventional and improved hens were detected in terms of egg-laying rate, fertilization rate, and hatchability in the Kyushu Rhode hens. The data suggest that fattening chicks can be supplied as usual, even if Kyushu Rhode hens are switched from the conventional to improved type. In conclusion, genetic improvements using the CCKAR SNP site as a marker were effectively established in terms of the growth of the Miyazaki Jitokko chickens in commercial farms and the reproductive traits of the Kyushu Rhode hens.