Development of a highly sensitive and rapid detection method for Pea enation mosaic virus using loop-mediated isothermal amplification assay.

The Pea enation mosaic virus (PEMV) has infected plants in the family Leguminosae such as pea, chickpea, faba bean, and lentil plants worldwide that the virus can be transmitted by sap, aphids, and seeds. Among the damages that PEMV disease cause in plants are reduced crop productivity, severely misshapen pods, wart-like outgrowths or proliferation on the surface. Previously, enzyme-linked immunosorbent assay (ELISA), reverse transcription (RT)-nested polymerase chain reaction (PCR), and real-time PCR had been used to detect PEMV. However, these methods are time-consuming and require specific equipments. For this reason, the development of a highly specific and sensitive detection method has become necessary. In this study, a new method for PEMV-1 using the loop-mediated isothermal amplification (LAMP) assay has been developed with specific primer sets as inner- and outer primers. Results showed PEMV-1 has been successfully detected that LAMP could confirm a diluted PEMV-1 up to 10-6 cDNA. LAMP is about 10,000 times more sensitive than the RT-nested PCR and/or real-time PCR. Moreover, the processing time of the LAMP was decreased 3 h than RT-nested PCR. Although future validation will be required to confirm enablement in the field area, this study provides a valuable method to identify PEMV-1 that could offer some advantages including rapid detection, high specificity and high sensitivity than others.

Mucin gene polymorphisms are associated with endometriosis in Korean women.

OBJECTIVE: Mucin family members mucin 1 (MUC1) and mucin 4 (MUC4) play an important role in transformation and adhesion, and are known markers for the detection of cancer. However, the pathophysiology of endometriosis associated with the mucin gene is unclear. In this study, we analyzed the relationship between MUC1 and MUC4 single-nucleotide polymorphisms (SNPs) and the risk for endometriosis. METHODS: We performed a case-controlled study of 29 endometriosis clinical samples and 27 functional cysts as control. Sixteen SNPs (rs145224844, rs139620330, rs144273480, rs1611770, rs146141676, rs201798179, rs201815857, rs199840128, rs200788986, rs141460657, rs183700327, rs199768496, rs191544901, rs200639498, rs148332231, and rs11465209) of MUC1 gene and eight SNPs (rs1104760, rs1106502, rs882605, rs2291651, rs2291652, rs2291653, rs2291654, and rs375068067) of the MUC4 gene were identified. We amplified SNP sites by polymerase chain reaction (PCR) using specific primer sets followed by DNA sequencing. RESULTS: The single mutation analysis of MUC4 showed that MUC4 mutations had no effect on the risk for endometriosis, but the frequencies of haplotypes [T/T + T/T + C/C] (rs2291653, 2291654 and rs375068067) were associated with endometriosis. CONCLUSION: The MUC1 genotype may not be correlated with endometriosis susceptibility. However, MUC4 polymorphisms are associated with the risk for endometriosis in Korean women.

Roles of 5-HT on phase transition of neurite outgrowth in the identified serotoninergic neuron C1, Helisoma trivolvis.

Neurite outgrowth is a morphological marker of neuronal differentiation and neuroregeneration, and the process includes four essential phases, namely initiation, elongation, guidance and cessation. Intrinsic and extrinsic signaling molecules seem to involve morphological changes of neurite outgrowth via various cellular signaling cascades phase transition. Although mechanisms associated with neurite outgrowth have been studied extensively, little is known about how phase transition is regulated during neurite outgrowth. 5-HT has long been studied with regard to its relationship to neurite outgrowth in invertebrate and vertebrate culture systems, and many studies have suggested 5-HT inhibits neurite elongation and growth cone motility, in particular, at the growing parts of neurite such as growth cones and filopodia. However, the underlying mechanisms need to be investigated. In this study, we investigated roles of 5-HT on neurite outgrowth using single serotonergic neurons C1 isolated from Helisoma trivolvis. We observed that 5-HT delayed phase transitions from initiation to elongation of neurite outgrowth. This study for the first time demonstrated that 5-HT has a critical role in phase-controlling mechanisms of neurite outgrowth in neuronal cell cultures.

Characterization of Impaired Cerebrovascular Structure in APP/PS1 Mouse Brains.

Alzheimer's disease (AD) is defined by senile plaques, tauopathy and neuronal cell death in specific area of the brain. Recent studies suggest that neurovascular dysfunction may be an integral part of AD pathogenesis, contributing to the onset and development of AD pathologies such as neuronal death, inflammatory response, and breakdown of blood-brain barrier (BBB). In addition, vascular complications caused by age-related metabolic diseases such as diabetes and high blood pressure have high incidence in development of dementia and AD. We previously reported that astrocytes, essential components of BBB, were chronically activated and some deteriorated in the brain of 5xFAD, an amyloid precursor protein/presenilin1 (APP/PS1) transgenic mouse model. Thus, it is rational to investigate if any vascular dysfunction is associated with considerable activation of astrocytes in APP/PS1 mouse model. In this study, we observed that cerebrovascular pathology was associated with large scale of reactive astrocytes and neurodegeneration in an Aß plague-generating mouse model. Using 5xFAD mouse brains, we demonstrate damaged brain vessels and reduced expression of glucose transporter 1 (GLUT1), the main glucose transporter, and a tight junction protein zonula occludens-1 (ZO-1) of cerebrovascular endothelial cells. This vascular pathology was closely associated with astrocytic deterioration and neuronal loss due to buildup of Aß plaques in 5xFAD mouse brains.

Polymorphisms of small ubiquitin-related modifier genes are associated with risk of Alzheimer's disease in Korean: A case-control study.

Sumoylation regulates transcription factor transactivation, protein-protein interactions, and appropriate subcellular localization of certain proteins. Previous studies have shown that sumoylation of amyloid precursor protein (APP) is associated with decreased levels of amyloid beta (Aß) proteins, suggesting that sumoylation may play a role in the pathogenesis of Alzheimer's disease (AD). We investigated the association between polymorphisms of the SUMO genes and the risk of AD. Our study subjects consisted of 144 AD patients and 335 healthy controls without dementia. We focused on tagged single nucleotide polymorphisms (tagSNPs) of the SUMO1 and SUMO2 genes. The tagSNPs were amplified by PCR and sequenced. We used binary logistic regression to calculate odds ratios (ORs) with 95% confidence intervals (CIs) for the associations between SUMO gene polymorphisms and the risk of AD. We found that rs12472035 polymorphism of SUMO1 was significantly associated with an increased risk of AD in male group (the CT genotype of rs12472035: adjusted OR=8.737, 95% CI=2.041-37.41, p-value=0.003). In addition, two polymorphisms of SUMO2 were significantly associated with an increased risk of AD in female group (the GA genotype of rs35271045: adjusted OR=2.879, 95% CI=1.399-5.924, p-value=0.004; and the TC genotype of rs9913676: adjusted OR=2.460, 95% CI=1.197-5.057, p-value=0.014). Furthermore, three combinations were associated with an increased risk of AD. Our data suggest that three individual polymorphisms and three combinations may be potential risk factors for AD in Korean population.

Ionic and thermo-switchable polymer-masked mesoporous silica drug-nanocarrier: High drug loading capacity at 10°C and fast drug release completion at 40°C.

The preparation of the ideal smart drug-delivery systems were successfully achieved by the in situ co-polymerization of a vinyl group-functionalized mesoporous silica nanoparticle (f-MSN) with 1-butyl-3-vinyl imidazolium bromide (BVIm) and N-isopropylacrylamide (NIPAAm) monomers. The thickness of the capping copolymer layer, poly(NIPAAm-co-BVIm) (p-NIBIm), was controlled at between 2.5nm and 5nm, depending on the monomers/f-MSN ratio in the reaction solution. The finally obtained smart drug-delivery systems are named as p-MSN2.5 and p-MSN5.0 (MSNs integrated by 2.5nm and 5nm p-NIBIm layer in thickness). The key roles of the mesoporous-silica-nanoparticle (MSN) core and the p-NIBIm shell are drug-carrying (or containing) and pore-capping, respectively, and the latter has an on/off function that operates in accordance with temperature changes. According to the swelling- or shrinking-responses of the smart capping copolymer to temperature changes between 10°C and 40°C, the loading and releasing patterns of the model drug cytochrome c were studied in vitro. The developed system showed interesting performances such as a cytochrome-c-loading profile (loading capacity for 3h=26.3% and 19.8% for p-MSN2.5 and p-MSN5.0, respectively) at 10°C and a cytochrome-c-releasing profile (releasing efficiency=>95% within 3 days and 4 days for p-MSN2.5 and p-MSN5.0, respectively) at 40°C. The cytotoxicity of the drug delivery systems, p-MSN2.5 and p-MSN5.0 (in the concentration range of <0.125mg/mL without drug), for human embryonic kidney (HEK 293) cells were minimal in vitro compared with that of a blank MSN. These results may be reasonably applied in the field of specified drug delivery.

Calcium homeostasis modulator 1 gene P86L polymorphism and the risk for alzheimer's disease: A meta-analysis.

OBJECTIVES: Recently, many epidemiological studies have demonstrated an association between P86L polymorphism of calcium homeostasis modulator 1 (CALHM1) and risk for Alzheimer's disease (AD). However, the results of these association studies are inconsistent. In this study, we re-evaluated the relation between CALHM1 P86L polymorphism and risk for AD in a meta-analysis. METHODS: This meta-analysis was performed using the PubMed, Science Direct, Scopus and Google Scholar databases up to June 2015 using the search terms "CALHM1" and "polymorphism or SNP or variant" in combination with "Alzheimer's disease". A meta-analysis with pooled odds ratios and 95% confidence intervals was carried out to assess the associations between P86L polymorphism and the risks for Alzheimer's disease under four genetic models with fixed or random effects models. RESULTS: Sixteen studies (twenty-four subgroup studies involving 9795 cases and 15,335 controls) were included in our meta-analysis. Our meta-analysis results indicated that several genetic models of CALHM1 P86L polymorphism were significantly associated with increased risk for AD in overall and Caucasian populations. CONCLUSIONS: In conclusion, our comprehensive meta-analysis indicated that P86L polymorphism is significantly associated with an increased risk for AD. Our data suggest that CALHM1 polymorphism may be potential biomarker in patients with AD.

Genetic polymorphisms of interleukin genes and the risk of Alzheimer's disease: An update meta-analysis.

OBJECTIVES: Recently, several meta-analyses have reported an association between interleukin (IL) gene polymorphisms and the risk of Alzheimer's disease (AD). Several further papers discussing the relationship with the risk of AD have recently been published. The aim of this meta-analysis was to re-evaluate and update the associations between IL gene polymorphisms and the risk of AD. METHODS: The search sources were PubMed, Science Direct, Scopus, and Google Scholar up to July 2015, and the following search terms were used: "interleukin 1 or interleukin 6 or interleukin 10" and "variant or polymorphism or SNP" in combination with "Alzheimer's disease". A meta-analysis using the pooled odds ratios and 95% confidence intervals was carried out to assess the associations between four polymorphisms of IL genes (- 889C > T in IL-1α, - 511C > T in IL-1ß, - 174G > C in IL-6 and - 1082G > A in IL-10) and the risk of AD under the heterozygous, homozygous, dominant, and recessive models with fixed- or random-effects models. RESULTS: A total of 21,864 cases and 40,321 controls from 93 individual studies were included in this meta-analysis. Our results indicated that the - 889C > T polymorphism was strongly associated with the increased risk of AD. However, three polymorphisms were not associated with the risk of AD. CONCLUSIONS: Similar to previous meta-analyses, our updated meta-analysis suggested that the - 889C > T polymorphism may be a factor in AD. However, the results of our meta-analysis of the - 174G > C polymorphism differed from those of previous meta-analyses. Consequently, we suggest that the - 174G > C polymorphism may not be a risk factor for AD.

Loop-mediated Isothermal Amplification Assay to Rapidly Detect Wheat Streak Mosaic Virus in Quarantined Plants.

We developed a loop-mediated isothermal amplification (LAMP) method to rapidly diagnose Wheat streak mosaic virus (WSMV) during quarantine inspections of imported wheat, corn, oats, and millet. The LAMP method was developed as a plant quarantine inspection method for the first time, and its simplicity, quickness, specificity and sensitivity were verified compared to current reverse transcription-polymerase chain reaction (RT-PCR) and nested PCR quarantine methods. We were able to quickly screen for WSMV at quarantine sites with many test samples; thus, this method is expected to contribute to plant quarantine inspections.

Vitamin D receptor gene polymorphisms and the risk for female reproductive cancers: A meta-analysis.

Vitamin D receptor (VDR) gene polymorphisms and the risks for various breast and ovarian cancers have been reported in many epidemiological studies. However, the associations between VDR gene polymorphisms and the risk for each type of cancer are unclear. The aim of this meta-analysis was to evaluate the associations between VDR gene polymorphisms and female reproductive cancers. A systematic review was performed with the PubMed Science Direct, Scopus, and Google Scholar databases up to April 2014 using the search terms "vitamin D receptor or VDR" and "variant or polymorphism or SNP" with terms for breast, ovarian, cervical, endometrial, uterine, and vaginal cancers. A meta-analysis with the pooled odds ratios and 95% confidence intervals was carried out to assess the associations between VDR polymorphisms (Cdx-2, FokI, BsmI, ApaI, and TaqI) and the risks for reproductive cancers under the heterozygous, homozygous, dominant, and recessive models with fixed or random effects models. Six ovarian cancer studies (13 individual studies involving 4107 cases and 6661 controls) and 29 breast cancer studies (38 individual studies involving 16,453 cases and 22,044 controls) were included in our meta-analysis. Our results indicate that the FokI polymorphism was related to increased risks for breast and ovarian cancers, whereas the BsmI polymorphism was associated with a decreased risk for developing these cancers. Our comprehensive meta-analysis indicated that the FokI and BsmI VDR gene polymorphisms may be significantly associated with gynecological cancers. We suggest monitoring VDR gene polymorphisms as potential biomarkers in patients with gynecological malignancy.

Biofunctionalized carbon nanotubes in neural regeneration: a mini-review.

Carbon nanotubes (CNTs) have become an intriguing and promising biomaterial platform for the regeneration and functional recovery of damaged nerve tissues. The unique electrical, structural and mechanical properties, diversity of available surface chemistry and cell-penetrating ability of CNTs have made them useful implantable matrices or carriers for the delivery of therapeutic molecules. Although there are still challenges being faced in the clinical applications of CNTs mainly due to their toxicity, many studies to overcome this issue have been published. Modification of CNTs with chemical groups to ensure their dissolution in aqueous media is one possible solution. Functionalization of CNTs with biologically relevant and effective molecules (biofunctionalization) is also a promising strategy to provide better biocompatibility and selectivity for neural regeneration. Here, we review recent advances in the use of CNTs to promote neural regeneration.

Promoter -2518 single nucleotide polymorphism of monocyte chemoattractant protein-1 is associated with clinical severity in Behçet's disease.

OBJECTIVE: We investigated whether promoter -2518 single nucleotide polymorphism (SNP) of the monocyte chemoattractant protein-1 (MCP-1) gene contributes to susceptibility and clinical features or severity in Behçet's disease (BD) patients. METHODS: One hundred and thirty-two BD patients and 113 healthy subjects, matched by sex and age, were enrolled. Promoter -2518 polymorphism of the MCP-1 gene was analyzed using automated sequencing. Clinical severity in BD patients was classified into mild, moderate, and severe features and assessed by total severity scores. Clinical features and severity was also compared according to genotypes using either the chi-squared or Fisher's exact test and Mann-Whitney test, as indicated. RESULTS: There were no significant differences in alleles (G allele vs. A allele, p=0.845) and genotypes with -2518 SNP (GG vs. GA vs. AA, p=0.916) between BD patients and controls. No clinical features were associated with genotypes with -2518 polymorphism of MCP-1. However, the frequency of either GA or AA genotype in patients with moderate lesions and moderate to severe lesions was significantly increased compared with that in patients with the GG genotype (p=0.044 and p=0.038, respectively). Total severity scores in the AA genotype were higher than those in the GG and GA genotypes (p=0.039 and p=0.003, respectively). Moreover, patients with either the GA or AA genotype had higher scores than those with the GG genotype (p=0.041). CONCLUSIONS: This study demonstrated that genotypes with A allele with -2518 polymorphism of the MCP-1 gene might have increased risk of severity of clinical features, but not susceptibility to BD.

G109T polymorphism of SLC22A12 gene is associated with serum uric acid level, but not with metabolic syndrome.

SLC22A12 gene, encoding urate transport 1, has been known to be responsible to urate metabolism. This study sought to determine the association between the novel G109T polymorphism in SLC22A12 with serum uric acid and the development of metabolic syndrome in Korean male subjects. A total of 132 healthy male subjects were enrolled in this study. Metabolic syndrome was determined using the modified guidelines for metabolic syndrome proposed by the National Cholesterol Education Program's Third Adult Treatment Panel. Genotyping for the SLC22A12 gene was assessed using denaturing high-performance liquid chromatography analysis. Serum uric acid and fractional excretion of uric acid (FEUA) from blood and urine samples were measured. Frequencies of the 109GG, 109GT, and 109TT genotypes were 57.6, 38.6, and 3.8%, respectively. Serum uric acid levels and FEUAs were significantly different among the three genotypes of the G109T polymorphism (P = 0.035 and P = 0.033, respectively). In addition, subjects of genotypes with the T allele had lower uric acid levels and higher FEUAs compared to those with the 109GG genotype (P = 0.007 and P = 0.031, respectively). The G109T polymorphism of the SLC22A12 gene has no association with metabolic syndrome. However, a number of metabolic syndrome components were related to serum uric acid level (r = 0.285, P = 0.001) and also significantly different between genotype with and without T allele (P = 0.008). The novel G109T polymorphism of the SLC22A12 gene is related to serum uric acid level, but not to the development of metabolic syndrome.

Carbon nanotubes in nanocomposites and hybrids with hydroxyapatite for bone replacements.

Hydroxyapatite (HA), as a bone mineral component, has been an attractive bioceramic for the reconstruction of hard tissues. However, its poor mechanical properties, including low fracture toughness and tensile strength, have been a significant challenge to the application of HA for the replacement of load-bearing and/or large bone defects. Among materials studied to reinforce HA, carbon nanotubes (CNTs: single-walled or multiwalled) have recently gained significant attention because of their unprecedented mechanical properties (high strength and toughness) and physicochemical properties (high surface area, electrical and thermal conductivity, and low weight). Here, we review recent studies of the organization of HA-CNTs at the nanoscale, with a particular emphasis on the functionalization of CNTs and their dispersion within an HA matrix and induction of HA mineralization. The organization of CNTs and HA implemented at the nanoscale can further be developed in the form of coatings, nanocomposites, and hybrid powders to enable potential applications in hard tissue reconstruction.

Polymorphisms of tumor necrosis factor-α promoter region for susceptibility to HLA-B27-positive ankylosing spondylitis in Korean population.

This study designed to assess the relationship between tumor necrosis factor (TNF)-α promoter polymorphisms and disease susceptibility to human leukocyte antigen (HLA)-B27-positive ankylosing spondylitis (AS). One hundred and nineteen HLA-B27(+) AS patients, 95 HLA-B27(+) healthy controls, and 135 random healthy controls were enrolled in this study. Six single nucleotide polymorphisms (SNPs) of the TNF-α promoter at positions -1031T/C, -863C/A, -857C/T, -646G/A, -308G/A, and -238G/A were analyzed. Differences between groups were evaluated using the chi-square test or Fisher's exact test. Haplotypes from each SNP were constructed, and differences in haplotypic frequencies between groups were evaluated. There were significant differences in the allelic and genotypic frequencies of 1031T/C, -863C/A, and -857C/T TNF-α promoters polymorphisms between HLA-B27(+) AS patients and random controls, but not between patients with AS and HLA-B27(+) healthy individuals. TNF-α polymorphisms did not influence the extra-spinal clinical features in patients with AS. The haplotypic sequence -1031T/-863C/-857C/-308G increased the risk of susceptibility to AS compared to random controls (P (corr) < 0.001, OR = 2.756, 95% CI = 1.894-4.010), whereas the sequence -1031C/-863A/-857C/-308G appeared to be associated with decreased susceptibility to AS compared to random controls (P (corr) = 0.006, OR = 0.396, 95% CI = 0.231-0.679). This study indicates that TNF-α promoter polymorphism between controls and AS patients with HLA-B27(+) genetic background is not associated with susceptibility to AS. However, TNF-α polymorphism, irrespective of HLA-B27, increases risk of susceptibility to AS in general population.

Relationship between salivary Chromogranin-A and stress induced by simulated monotonous driving.

The purpose of the present study was to evaluate the use of salivary Chromogranin-A (CgA), which is already used in general as a mental stress marker, for studying the stressful situation created by simulated monotonous driving. After informed consent, 25 healthy male and female subjects were studied under constant environment-controlled conditions. We measured the following physiological variables: blood pressure (BP), cardiac output, total peripheral resistance (TPR), normalized pulse volume (NPV) as an index of alpha-adrenergic sympathetic activity to the finger arteriolar vessels, levels of cortisol and CgA during monotonous driving. The induced stress led to the expected decreases in NPV and increases in TPR and BP caused by peripherally related sympathetic acceleration. However, CgA levels were found to fall gradually in accordance with the gradual increase of subjective rating of stress (SRS) and significantly (p < 0.01) decreased over the period of the simulated monotonous driving. Our hypothesis for the gradual decrease of CgA levels during the simulated monotonous driving is as follows. CgA, catestatin and catecholamines are co-released into the extra-cellular environment. Peripheral sympathetic activity was accelerated by stress resulting from the simulated monotonous driving. Upon peripheral vessel constriction, an increase in TPR then increased BP which, in turn, activated catestatin. Consequently, secretion of CgA was blocked by the co-secreted catestatin from chromaffin granules. The results obtained strongly indicate that, although CgA has been reported as a possible marker of stress, CgA levels are not increased in the stressful situation of simulated monotonous driving.

T6092C polymorphism of SLC22A12 gene is associated with serum uric acid concentrations in Korean male subjects.

BACKGROUND AND OBJECT: It has been suggested that the SLC22A12 gene polymorphism may be involved in the mechanism of renal urate handling. The purpose of the current study was to identify the effect of the T6092C polymorphism of the SLC22A12 gene on serum uric acid concentrations in the Korean population. METHODS: We examined 196 healthy subjects (141 males and 55 females) in this study. Among the SLC22A12 gene polymorphism, the T6092C polymorphism in intron 4 at rs1529909 of the SLC22A12 gene was evaluated using denaturing high performance liquid chromatography analysis. Diverse clinical parameters of renal urate handling derived from fasting blood and urine samples, such as the serum uric acid concentration and the fractional excretion of uric acid (FEUA), were also assessed for identification of the relationship between genotypic variations. Statistical analysis was performed using the chi-square test, ANOVA test, and the Pearson correlation coefficient analysis to determine which indicators involved serum uric acid. And the significance of these indicators was then confirmed by multivariate regression analysis. RESULTS: The prevalence of the T6092C polymorphism (TT, TC, and CC) was 58.2%, 37.2%, and 4.6%, respectively. Only the concentration of serum uric acid and the FEUA in male subjects differed significantly among each genotype (p=0.038 and p=0.013, respectively). Serum uric acid concentrations in male subjects with the TT genotype were increased compared with those with the TC as well as the TC or CC genotypes (6.2+/-1.2 vs. 5.7+/-1.3, p=0.039 and 6.2+/-1.2 vs. 5.6+/-1.3, p=0.019, respectively), whereas there was no significant difference between CC genotype and TT genotype in serum uric acid (p=0.066). No significant difference between clinical indicators and genotypes existed in females. The T6092C polymorphism of the SLC22A12 gene, serum creatinine, and the FEUA were significantly associated with the concentration of serum uric acid. CONCLUSION: This study showed that the T6092C polymorphism of the SLC22A12 gene may be involved in renal urate handling and the concentration of serum uric acid, in male subjects.

Interleukin-17F gene polymorphisms in Korean patients with Behçet's disease.

IL-17 is a novel cytokine that is characterized by an ability to induce several types of cells to secrete proinflammatory cytokines in various inflammatory diseases. This study analyzed the influence of IL-17F gene polymorphisms on disease susceptibility and clinical features. Ninety-nine Behçet's disease (BD) patients and 114 controls were genotyped to analyze three single nucleotide polymorphisms (SNPs) including A126G, G155A, and A161G of the IL-17F gene using automated sequencing. We compared the frequencies of IL-17F alleles, genotypes, and haplotypes in patients with BD and controls using the chi-square or Fisher's exact test. Significant differences in the frequencies of allele and genotype in A126G SNP of IL-17 gene were found between BD patients and controls (P<0.001 and P<0.001, respectively). None of three IL-17F SNPs were associated with diverse clinical features in BD. The frequency of haplotype AA did not differ between patients with BD and controls (P=0.985). The haplotypes, AG, and GG, have positive and inverse association with BD susceptibility (P<0.001 and P<0.001, respectively). These findings suggest that IL-17 gene SNPs may influence the susceptibility of BD.

[Detection of rifampin resistant mycobacterium tuberculosis complex using denaturing HPLC].

BACKGROUND: Tuberculosis (TB) remains an important cause of morbidity and mortality throughout the world. The surge of TB has been accompanied by an increase in multi-drug-resistant tuberculosis (MDR-TB). In this study, we developed a denaturing HPLC (DHPLC) method for detecting rpoB gene mutation as a rifampin resistance based on sequence. METHODS: In this study, we used 99 mycobacterial isolates grown in Ogawa media. At first, we used a PCR method that can amplify the 235 bp and 136 bp rpoB DNAs of Mycobacterium tuberculosis complex (MTB) and Non-tuberculous mycobacteria (NTM). And then, PCR-restriction fragment length polymorphism (RFLP) of rpoB DNA (342 bp), which comprises the Rif(T) region, was used for the differential identification of Mycobacteria. Finally, we detected these amplicons by DHPLC, compared to PCR-RFLP results, and performed sequencing. RESULTS: Among 99 mycobacterial isolates, 80 (81%) were MTB and 19 (19%) were NTM. NTM were identified to 7 different species by DHPLC and PCR-RFLP. rpoB mutation was detected in 9 (11%) of the MTB specimens. These results were confirmed by using sequencing. CONCLUSIONS: DHPLC provided a rapid, simple, and automatable performance for detection of rifampin resistant Mycobacterium tuberculosis complex and would be helpful as a supplemental method in high-throughput clinical laboratories.

A novel mutation in the SCN5A gene is associated with Brugada syndrome.

Brugada syndrome (BS) is an inherited cardiac disorder associated with a high risk of sudden cardiac death and is caused by mutations in the SCN5A gene encoding the cardiac sodium channel alpha-subunit (Na(v)1.5). The aim of this study was to identify the genetic cause of familial BS and characterize the electrophysiological properties of a novel SCN5A mutation (W1191X). Four families and one patient with BS were screened for SCN5A mutations by PCR and direct sequencing. Wild-type (WT) and mutant Na(v)1.5 channels were expressed in tsA201 cells, and the sodium currents (I(Na)) were analyzed using the whole-cell patch-clamp technique. A novel mutation, W1191X, was identified in a family with BS. Expression of the WT or the mutant channel (Na(v)1.5/W1191X) co-transfected with the beta(1)-subunit in tsA201 cells resulted in a loss of function of Na(v)1.5 channels. While voltage-clamp recordings of the WT channel showed a distinct acceleration of Na(v)1.5 activation and fast inactivation kinetics, the Na(v)1.5/W1191X mutant failed to generate any currents. Co-expression of the WT channel and the mutant channel resulted in a 50% reduction in I(Na). No effect on activation and inactivation were observed with this heterozygous expression. The W1191X mutation is associated with BS and resulted in the loss of function of the cardiac sodium channel.