Genetic associations between miR­200bT>C and miR­495A>C polymorphisms and hypertension susceptibility.

Although microRNA (miRNA)-mediated functions and gene expression regulation are involved in the susceptibility to vascular diseases, the potential effect of miRNA polymorphisms on the susceptibility of patients to hypertension (HTN) remains to be sufficiently elucidated. Therefore, the present study aimed to identify the potential association between miRNA (miR)-200bT>C (rs7549819) and miR-495A>C (rs2281611) polymorphisms, which may be implicated in stroke and vascular pathogenesis, and the susceptibility to HTN and relevant risk factors in a Korean cohort recruited from Jeju National University Hospital (Jeju, South Korea). Using an analysis of PCR-restriction fragment length polymorphism, genotype analysis was conducted to assess the frequency of miR-200bT>C and miR-495A>C gene polymorphisms in the HTN group (n=232) and the non-HTN healthy control group (n=247). The results showed significant differences in the genotype distributions of the miR-495A>C polymorphism between the HTN and control groups, specifically with the CC genotype and C allele. However, neither the miR-200bT>C nor the dominant and recessive models were found to be distributed differently between the two groups. Following analysis of the genotype combination of the single nucleotide polymorphisms, the TC/CC and CC/CC combined genotypes of the miR-200bT>C and miR-495A>C polymorphisms were observed to be associated with susceptibility to HTN. The haplotype results demonstrated that the allele combination frequency of haplotype C-A was significantly different between the two groups. The stratified analysis revealed that the miR-200b and miR-495 polymorphisms are associated with the risk of HTN, exhibiting differences in the levels of body-mass index (<28.12 kg/m2), fasting blood glucose (<106.26 mg/dl), high-density lipoprotein cholesterol (<44.29 mg/dl) and systolic blood pressure (≥132.67 mmHg). Data from the present study suggested that the variant of miR-495A>C polymorphism and allelic combinations (haplotype C-A of miR-200bT>C/miR-495A>C) can increase hypertension susceptibility among a Korean population.

Theory of mind in children with cochlear implants: Comparison with age- and sex-matched children with normal hearing.

OBJECTIVES/HYPOTHESIS: Theory of mind (ToM) is a crucial ability for maintaining normal social interaction and is directly related to language ability. This study was performed to compare ToM between children with congenital hearing loss who have received cochlear implantation (CI) and those with normal hearing (NH). STUDY DESIGN: Case-control study design. METHODS: One hundred children, aged 2-12 years, participated: 50 children who received CI before 36 months of age (CI group) and one-to-one age- and sex-matched children with normal hearing (NH group). All children underwent tests to examine receptive language ability and ToM. Receptive language was measured using the Receptive and Expressive Vocabulary Test, and ToM was measured using the Theory of Mind Task Battery (ToM-TB). The scores of the two tests were compared between the CI and NH groups. RESULTS: The ToM-TB score in the CI group correlated positively with age and receptive language score. ToM-TB scores did not differ significantly between children in the CI group who achieved normal receptive language and the NH group. However, these children in the CI group scored lower than those in the NH group on some advanced ToM tasks that require the ability to understand second-order emotion, message-desire discrepancy, or second-order false belief. CONCLUSIONS: This case-control study found that children with CI who achieve normal receptive language ability have ToM that is similar to that in children with NH. However, these children exhibited weakness in advanced ToM skills. Interventions to facilitate the development of advanced ToM are needed for children with CI.

Hyaluronan Synthase 1: A Novel Candidate Gene Associated With Late-Onset Non-syndromic Hereditary Hearing Loss.

OBJECTIVES: Hyaluronan synthase 1 (HAS1) is a membrane-bound protein that is abundant in the epidermis and dermis, and it is important for skin function. However, its association with hearing loss has not yet been studied. Herein, we sought to evaluate the potential contribution of HAS1: c.1082G>A to genetic hearing loss. METHODS: We used whole-exome sequencing to analyze blood DNA samples of six patients of a family with autosomal dominant familial late-onset progressive hearing loss, which was revealed to be related to a variant of the HAS1 gene. Confirmatory Sanger sequencing was performed with samples from 10 members. A missense variant was detected in HAS1 (c.1082 G>A, p.Cys361Tyr). In silico analyses predicted this variant to result in the functional loss of HAS1. Immunostaining was conducted using wild-type mouse samples to verify HAS1 expression. RESULTS: Has1 was detected in an otocyst at E10.5. In the pup, Has1 expression was localized in the stria vascularis (SV), hair cells, supporting cells of the organ of Corti, and some spiral ganglion neurons. SV marginal cells markedly expressed Has1 in the adult stage. The hearing threshold in the Has1-depleted condition was investigated by accessing the International Mouse Phenotyping Consortium's Auditory Brainstem Response (ABR) data. ABR of Has1 knock-out mice showed threshold elevations at 6, 12, and 18 kHz in young male adults. CONCLUSION: HAS1 may have a close relationship with auditory function and genetic hearing loss. Further investigation is needed to reveal the precise role of HAS1 in the auditory system. HAS1 is a candidate gene for future hereditary hearing loss genetic testing.

Genetic association between miR-27a and miR-449b polymorphisms and susceptibility to diabetes mellitus.

Various mutations in microRNAs (miRs) are associated with the pathogenesis of several diseases including cancers and vascular diseases. The present study aimed to investigate the potential association between miR-27a A>G (rs895819) and miR-449b A>G (rs10061133) polymorphisms with the prevalence of type 2 diabetes mellitus (T2DM), and its associated risk factors in the Korean population. Genotype analysis was performed using PCR-restriction fragment length polymorphism analysis to assess the frequency of miR-27a and miR-449b gene polymorphisms in patients diagnosed with T2DM (n=238) and healthy controls (n=247). The miR-27a GG genotype, recessive model, and G allele were significantly associated with a decreased risk of T2DM [adjusted odds ratio (AOR)=0.378, 95% confidence interval (CI): 0.208-0.686, P=0.001; AOR=0.425, 95% CI: 0.246-0.734, P=0.002; AOR=0.640, 95% CI: 0.493-0.831, P=0.001, respectively). Although the miR-449b polymorphism was not associated with the prevalence of T2DM, the genotype and allele combination analyses for miR-27a and miR-449b polymorphisms showed associations with T2DM prevalence. Furthermore, stratification analysis revealed that the miR-27a polymorphism was associated with DM risk factors including body mass index (<28.12 kg/m2, P=0.031), waist circumference (<93.03 cm, P=0.036), systolic blood pressure (<132.67 mmHg, P=0.017), fasting blood glucose levels (<106.26 mg/dl, P=0.015), glycosylated hemoglobin, type A1C (≤125.5 mg/dl, P=0.001), total cholesterol (≤240 mg/dl, P=0.010) and low-density lipoprotein levels (≤130 mg/dl, P=0.028). The present study revealed an association between miR-27a A>G and miR-449b A>G polymorphisms and the risk of DM in Koreans, which suggests that these gene polymorphisms could represent potential markers for predicting T2DM risk.

Efficiency of antioxidant Avenanthramide-C on high-dose methotrexate-induced ototoxicity in mice.

Methotrexate (MTX) has been used in treating various types of cancers but can also cause damage to normal organs and cell types. Folinic acid (FA) is a well-known MTX antidote that protects against toxicity caused by the drug and has been used for decades. Since hearing loss caused by MTX treatment is not well studied, herein we aimed to investigate the efficiency of the antioxidant Avenanthramide-C (AVN-C) on high-dose MTX (HDMTX) toxicity in the ear and provide insights into the possible mechanism involved in MTX-induced hearing loss in normal adult C57Bl/6 mice and HEI-OC1 cells. Our results show that the levels of MTX increased in the serum and perilymph 30 minutes after systemic administration. MTX increased hearing thresholds in mice, whereas AVN-C and FA preserved hearing within the normal range. MTX also caused a decrease in wave I amplitude, while AVN-C and FA maintained it at higher levels. MTX considerably damaged the cochlear synapses and neuronal integrity, and both AVN-C and FA rescued the synapses. MTX reduced the cell viability and increased the reactive oxygen species (ROS) level in HEI-OC1 cells, but AVN-C and FA reversed these changes. Apoptosis- and ROS-related genes were significantly upregulated in MTX-treated HEI-OC1 cells; however, they were downregulated by AVN-C and FA treatment. We show that MTX can cause severe hearing loss; it can cross the blood-labyrinth barrier and cause damage to the cochlear neurons and outer hair cells (OHCs). The antioxidant AVN-C exerts a strong protective effect against MTX-induced ototoxicity and preserved the inner ear structures (synapses, neurons, and OHCs) from MTX-induced damage. The mechanism of AVN-C against MTX suggests that ROS is involved in HDMTX-induced ototoxicity.

Development of DNA markers for Slmlo1.1, a new mutant allele of the powdery mildew resistance gene SlMlo1 in tomato (Solanum lycopersicum).

Reductions in growth and quality due to powdery mildew (PM) disease cause significant economic losses in tomato production. Oidium neolycopersici was identified as the fungal species responsible for tomato PM disease in South Korea in the present study, based on morphological and internal transcribed spacer DNA sequence analyses of PM samples collected from two remote regions (Muju and Miryang). The genes involved in resistance to this pathogen in the tomato accession 'KNU-12' (Solanum lycopersicum var. cerasiforme) were evaluated, and the inheritance of PM resistance in 'KNU-12' was found to be conferred via simple Mendelian inheritance of a mutant allele of the PM susceptibility locus Ol-2 (SlMlo1). Full-length cDNA analysis of this newly identified mutant allele (Slmlo1.1) showed that a 1-bp deletion in its coding region led to a frameshift mutation possibly resulting in SlMlo1 loss-of-function. An alternatively spliced transcript of Slmlo1.1 was observed in the cDNA sequences of 'KNU-12', but its direct influence on PM resistance is unclear. A derived cleaved amplified polymorphic sequence (dCAPS) and a high-resolution melting (HRM) marker were developed based on the 1-bp deletion in Slmlo1.1, and could be used for efficient marker-assisted selection (MAS) using 'KNU-12' as the source for durable and broad-spectrum resistance to PM.

Are the adverse effects of body position in patients with obstructive sleep apnea dependent on sleep stage?

PURPOSE: The purpose of the study was to determine if the adverse effect of body position on obstructive sleep apnea (OSA) is worsened during rapid eye movement (REM) sleep and if patients with OSA decrease the time spent supine during REM sleep. METHODS: Overnight polysomnography from 80 sequential patients referred to Buffalo VA Sleep Lab for suspected OSA were analyzed with 20 patients in each of the following groups: normal with apnea-hypopnea indices (AHI) <5/h, mild (AHI, 5-< 15/h), moderate (AHI, 15-<30/h), and severe (AHI, >30/h). We used extended Cox models with the Anderson-Gill modification for multiple events with two time varying covariates: sleep stage and body position. Generalized estimating equations with logit link were used to take into account correlated data within each patient for the relation between sleep stage and body position. RESULTS: The hazard ratios for events in REM vs non-REM sleep was significant for the normal, mild, and moderate groups only: 1.71 (95% CI 1.4-2.08), 1.45 (95% CI 1.22-1.73), 1.28 (95% CI 1.1-1.5), respectively. The hazard ratio for events in the supine vs non-supine position was significant for the mild and moderate groups only: 1.25 (95% CI 1.02-1.52) and 1.24 (95% CI 1.04-1.47), respectively. The addition of an interaction effect between sleep stage and body position was not statistically significant for any group. The odds ratios of sleeping in supine position for REM vs non-REM sleep were 0.47 (95% CI 0.27-0.82) for moderate OSA group and 0.54 (95% CI 0.3-0.95) for severe OSA. CONCLUSION: In summary, we found significant effects of both sleep stage and body position in mild and moderate but not severe OSA. Patients with moderate and severe OSA were less likely to spend time in the supine position during REM compared with non-rapid eye movement sleep.

Sleep-disordered breathing and chronic opioid therapy.

OBJECTIVE: To assess the relation between medications prescribed for chronic pain and sleep apnea. DESIGN: An observational study of chronic pain patients on opioid therapy who received overnight polysomnographies. Generalized linear models determined whether a dose relation exists between methadone, nonmethadone opioids, and benzodiazepines and the indices measuring sleep apnea. SETTING: A private clinic specializing in the treatment of chronic pain. PATIENTS: Polysomnography was sought for all consecutive (392) patients on around-the-clock opioid therapy for at least 6 months with a stable dose for at least 4 weeks. Of these, 147 polysomnographies were completed (189 patients declined, 56 were directed to other sleep laboratories by insurance companies, and data were incomplete for seven patients). Available data were analyzed on 140 patients. OUTCOME MEASURES: The apnea-hypopnea index to assess overall severity of sleep apnea and the central apnea index to assess central sleep apnea. RESULTS: The apnea-hypopnea index was abnormal (> or =5 per hour) in 75% of patients (39% had obstructive sleep apnea, 4% had sleep apnea of indeterminate type, 24% had central sleep apnea, and 8% had both central and obstructive sleep apnea); 25% had no sleep apnea. We found a direct relation between the apnea-hypopnea index and the daily dosage of methadone (P = 0.002) but not to other around-the-clock opioids. We found a direct relation between the central apnea index and the daily dosage of methadone (P = 0.008) and also with benzodiazepines (P = 0.004). CONCLUSIONS: Sleep-disordered breathing was common in chronic pain patients on opioids. The dose-response relation of sleep apnea to methadone and benzodiazepines calls for increased vigilance.

Asian plantain (Plantago asiatica) essential oils suppress 3-hydroxy-3-methyl-glutaryl-co-enzyme A reductase expression in vitro and in vivo and show hypocholesterolaemic properties in mice.

Asian plantain (Plantago asiatica) essential oil (PAEO) contains multiple bioactive compounds, but its potential effects on lipid metabolism have not been examined. PAEO was found to be mostly composed of oxygenated monoterpenes, with linalool as the major component (82.5 %, w/w), measured using GC-MS. Incubation of 0-200 microg PAEO/ml with HepG2 cells for 24 h resulted in no significant toxicity. Incubation with 0.2 mg PAEO/ml altered the expression of LDL receptor (+83 %; P < 0.05) and 3-hydroxy-3-methyl-glutaryl-CoA (HMG-CoA) reductase ( - 37 %; P < 0.05), as assessed using RT-PCR. LDL oxidation was markedly inhibited by PAEO treatment due to the prevalence of linalool compounds in PAEO. Oral administration of PAEO for 3 weeks in C57BL/6 mice significantly reduced plasma total cholesterol and TAG concentrations by 29 and 46 %, respectively. The mRNA (+58 %; P < 0.05), but not protein, levels of the LDL receptor were significantly higher, whereas both mRNA and protein levels of HMG-CoA reductase were significantly lower ( - 46 and - 11 %, respectively; P < 0.05) in the liver of PAEO-fed than of control mice. The mRNA levels of CYP7A1 were marginally reduced in HepG2 cells, but not in mouse liver after PAEO treatment. Thus, PAEO may have hypocholesterolaemic effects by altering the expression of HMG-CoA reductase. Reduced TAG and oxidised LDL may provide additional cardiovascular protective benefits.

Anopheles kleini, Anopheles pullus, and Anopheles sinensis: potential vectors of Plasmodium vivax in the Republic of Korea.

Anopheles sinensis Wiedemann (63.3%) was the most abundant Anopheles mosquito captured at cowshed resting collections in malaria high-risk areas (northern Gyeonggi Province) near the demilitarized zone (DMZ) in Korea during 2005, followed by Anopheles kleini Rueda (24.7%) and Anopheles pullus M. Yamada (8.7%). At cowshed resting collections in malaria low-risk areas (Jeonnam and Gyeongnam provinces), An. sinensis accounted for 96.8% of all Anopheles spp. collected, followed by An. kleini Rueda (2.7%), whereas no An. pullus were collected. Three species, An. kleini (50.9%), An. pullus (29.0%), and An. sinensis (13.8%), accounted for nearly all of the 224 Anopheles spp. captured by New Jersey light trap near the DMZ. In addition, An. pullus and An. kleini captured by New Jersey light trap near the DMZ and assayed by enzyme linked immunosorbent assay for Plasmodium vivax circumsporozoite antigen concentrations were higher than An. sinensis sensu stricto (s.s.), indicating higher levels of sporozoites. In laboratory studies of four concurrent artificial membrane feedings on malaria-infected blood from patients, F1 progeny of An. kleini and An. pullus had higher infection rates (8.8 and 7.5%, respectively) than An. sinensis s.s. (4.2%). These data suggest that An. kleini and An. pullus and An. sinensis are vectors of malaria in Korea. Further studies are required to determine the role of these species in the transmission of P. vivax in the Republic of Korea.

Coagulation parameters and plasma total homocysteine levels in Behcet's disease.

Behcet's disease (BD) is a multisystemic inflammatory disorder of unknown etiology that is sometimes associated with thrombosis in addition to systemic manifestations. However, the mechanism of hypercoagulability is not known. We evaluated the coagulation and fibrinolytic activities and the plasma total homocysteine levels of Korean BD patients in two cross-sectional studies. In the first study regarding coagulation and fibrinolytic activities, the levels of fibrinogen and von Willebrand factor (vWF) antigen were significantly higher in the BD patients than in the healthy controls (387.7+/-24.3 versus 240.6+/-8.8 mg/dl, p<0.001, and 131.9+/-8.8 versus 105.2+/-0.3%, p<0.01, respectively). The level of antithrombin III (AT III) was significantly lower in the BD patients (92.8+/-3.2 versus 106.3+/-2.6%, p<0.005). No differences were observed in the levels of plasminogen, protein C, or protein S activities. Activated protein C (APC) resistance was not observed in any BD patients. In the second study, the plasma total homocysteine levels of patients with a history of thrombosis (11.9+/-3.0 micromol/l) or disease activity (12.5+/-3.8 micromol/l) were found to be significantly higher than those of the controls (9.2+/-2.6 micromol/l, p<0.05, both). The plasma homocysteine concentrations in the thrombosis patients were positively correlated with plasma vWF levels; a relationship which suggests injury of the vascular endothelium (Spearman coefficient=0.857, p<0.01). Therefore, coagulation abnormality did not contribute to thrombotic complications, and higher levels of homocysteine may play a role in the hypercoagulablity of BD patients.