Full-length sequence of mouse acupuncture-induced 1-L (aig1l) gene including its transcriptional start site.

We have been investigating the molecular efficacy of electroacupuncture (EA), which is one type of acupuncture therapy. In our previous molecular biological study of acupuncture, we found an EA-induced gene, named acupuncture-induced 1-L (Aig1l), in mouse skeletal muscle. The aims of this study consisted of identification of the full-length cDNA sequence of Aig1l including the transcriptional start site, determination of the tissue distribution of Aig1l and analysis of the effect of EA on Aig1l gene expression. We determined the complete cDNA sequence including the transcriptional start site via cDNA cloning with the cap site hunting method. We then analyzed the tissue distribution of Aig1l by means of northern blot analysis and real-time quantitative polymerase chain reaction. We used the semiquantitative reverse transcriptase-polymerase chain reaction to examine the effect of EA on Aig1l gene expression. Our results showed that the complete cDNA sequence of Aig1l was 6073 bp long, and the putative protein consisted of 962 amino acids. All seven tissues that we analyzed expressed the Aig1l gene. In skeletal muscle, EA induced expression of the Aig1l gene, with high expression observed after 3 hours of EA. Our findings thus suggest that the Aig1l gene may play a key role in the molecular mechanisms of EA efficacy.

Evaluation of the Production of Dissolved Organic Matter by Three Marine Bacterial Strains.

A large part of marine dissolved organic matter (DOM) is considered to be recalcitrant DOM (RDOM) produced by marine bacteria. However, it is still unclear whether differences in bacterial species and/or physiology control the efficiency of RDOM production. Here, batch culture experiments with glucose as the sole carbon source were carried out using three model marine bacterial strains, namely, Alteromonas macleodii (Alt), Vibrio splendidus (Vib), and Phaeobacter gallaeciensis (Pha). Dissolved organic carbon (DOC) concentrations drastically decreased during the exponential growth phases of these bacteria due to the consumption of glucose. The efficiency of bacterial DOC production at the end of incubation was largely different among the strains and was higher for Vib (20%) than for the other two strains (Alt, 4%; Pha, 6%). All strains produced fluorescent DOM (FDOM), including humic-like FDOM which is considered as recalcitrant component in the ocean, even though the composition of bacterial FDOM was also different among the strains. The efficiency of humic-like FDOM production during the exponential growth phase was different among the bacterial strains; that is, Pha produced humic-like FDOM efficiently compared with the other two species. The efficiency of humic-like FDOM production with mineralization of organic matter was lower during the exponential growth phase than during the stationary phase of Alt and Pha. Four processes for the production of bacterially derived recalcitrant humic-like FDOM are suggested from this study: (1) production during active growing (in all strains), (2) production with the reutilization of bacterial DOM (Alt), (3) production with the consumption of cellular materials (Pha), and (4) release from lysis (Vib). Our results suggest that bacterial species and physiology can regulate RDOM production and accumulation in the ocean.

Pilsicainide Intoxication with Neuropsychiatric Symptoms Treated with Continuous Hemodiafiltration.

A 72-year-old lady with atrial fibrillation and chronic renal failure was hospitalized due to bradycardic shock with electrocardiographic QRS prolongation. She had experienced limb shaking two days before hospitalization, and additionally developed hallucinations one day before admission. Pilsicainide intoxication was diagnosed from a review of her medications and electrocardiographic findings. Consequently, continuous hemodiafiltration was performed resulting in a resolution of the hallucinations and the QRS prolongation. This is a rare case of psychiatric symptoms caused by pilsicainide intoxication. It is important to know the mode of excretion of a drug and to adjust its dose, so that such drug-related incidents can be avoided.

Interactive effect of paraoxonase-1 Q192R polymorphism and smoking history on the lung function decline in grain workers.

PURPOSE: This retrospective longitudinal study investigated the association between the Q192R polymorphism of the high-density lipoprotein-associated multifunctional antioxidant enzyme, paraoxonase-1 (PON1), and lung function decline, while taking into account smoking history. METHODS: The demographic, occupational, and respiratory symptom information and lung function variables were obtained from 216 male Saskatchewan grain workers. RESULTS: An interaction between the PON1 genotypes and smoking status was observed. Current smokers with the 192R allele had a lower forced expiratory volume in the first second (FEV(1)) and FEV(1) per forced vital capacity (FVC). The annual decline rate of FEV(1)/FVC in current smokers was greater among 192R allele carriers than noncarriers (0.58+/-0.05 vs. 0.35+/-0.04 %/yr, p<0.0001). A similar result was observed with FEV(1) (40.9+/-6.4 vs. -33.0+/-7.0 mL/yr, p=0.10). The annual decline rate of FVC was not influenced by the genotypes. CONCLUSIONS: These results strengthened the previous findings of our cross-sectional study, suggesting that the 192R allele may be a novel genetic risk factor for airway injury among current smokers.

Potential relationships between transaminase abnormality and valproic acid clearance or serum carnitine concentrations in Japanese epileptic patients.

This study tested the hypothesis that the determinants of mild liver injury are prerequisites for more severe idiosyncratic hepatotoxicity. This study verified whether the possible risk factors for rare idiosyncratic valproic acid (VPA)-induced hepatotoxicity, VPA clearance and/or serum carnitine concentrations are common to those for a mild elevation in transaminases in VPA-treated patients. VPA clearance was calculated in 172 Japanese patients with epilepsy, using a non-linear mixed-effects regression program. Carnitine concentrations were determined in a subset of 60 patients. The relationships between VPA clearance, carnitine concentration and levels of transaminases and ammonia were evaluated by Pearson's correlation coefficients. The final model of VPA apparent clearance (CL/F) was as follows: CL/F (L h(-1) = 0.012 x (BW/40)(0.34) x dose(0.55) x 0.90(gender) x 1.32(PHT) x 1.11(CBZ) x 1.12(PB), where BW = total body weight (kg); gender = 1 if female, 0 if male; PHT/CBZ/PB = 1 if phenytoin, carbamazepine, or phenobarbital, respectively, is coadministrated, otherwise 0. Either a higher VPA clearance or acyl/free carnitine ratio and a lower total and/or free carnitine concentration, but not VPA concentration, were associated with the mild elevation in transaminases or ammonia. These results support the initial hypothesis, while also helping to clarify the mechanism of severe idiosyncratic hepatotoxicity with VPA.

Electroacupuncture suppresses myostatin gene expression: cell proliferative reaction in mouse skeletal muscle.

Complementary and alternative medicine (CAM) may provide patients with an alternative to traditional medicine, but an assessment of its efficacy is required. One CAM method, electroacupuncture (EA) treatment, is a maneuver that utilizes stimulation of acupuncture needles with a low-frequency microcurrent. To study the effect of short-term EA, we evaluated the differential expression of genes induced by EA in mouse skeletal muscle for up to 24 h. We then used RT-PCR to confirm the expression patterns of six differentially expressed genes. Bioinformatics analysis of their transcription control regions showed that EA-inducible genes have numerous common binding motifs that are related to cell differentiation, cell proliferation, muscle repair, and hyperplasia. These results suggested that EA treatment may induce cell proliferation in skeletal muscle. To verify this possibility, we used EA to stimulate mouse skeletal muscle daily for up to 1 mo and examined the long-term effects. Immunohistochemical analysis showed that nuclei of muscle cells treated with EA for 1 mo, especially nuclei of satellite cells, reacted with anti-human PCNA. Also, expression of the gene encoding myostatin, which is a growth repressor in muscle satellite cells, was suppressed by daily EA treatment for 1 wk; EA treatment for 1 mo resulted in more marked suppression of the gene. These molecular findings constitute strong evidence that EA treatment suppresses myostatin expression, which leads to a satellite cell-related proliferative reaction and repair in skeletal muscle.

Production and Reutilization of Fluorescent Dissolved Organic Matter by a Marine Bacterial Strain, Alteromonas macleodii.

The recalcitrant fraction of marine dissolved organic matter (DOM) plays an important role in carbon storage on the earth's surface. Bacterial production of recalcitrant DOM (RDOM) has been proposed as a carbon sequestration process. It is still unclear whether bacterial physiology can affect RDOM production. In this study, we conducted a batch culture using the marine bacterial isolate Alteromonas macleodii, a ubiquitous gammaproteobacterium, to evaluate the linkage between bacterial growth and DOM production. Glucose (1 mmol C L-1) was used as the sole carbon source, and the bacterial number, the DOM concentration in terms of carbon, and the excitation-emission matrices (EEMs) of DOM were monitored during the 168-h incubation. The incubation period was partitioned into the exponential growth (0-24 h) and stationary phases (24-168 h) based on the growth curve. Although the DOM concentration decreased during the exponential growth phase due to glucose consumption, it remained stable during the stationary phase, corresponding to approximately 4% of the initial glucose in terms of carbon. Distinct fluorophores were not evident in the EEMs at the beginning of the incubation, but DOM produced by the strain exhibited five fluorescent peaks during exponential growth. Two fluorescent peaks were similar to protein-like fluorophores, while the others could be categorized as humic-like fluorophores. All fluorophores increased during the exponential growth phase. The tryptophan-like fluorophore decreased during the stationary phase, suggesting that the strain reused the large exopolymer. The tyrosine-like fluorophore seemed to be stable during the stationary phase, implying that the production of tyrosine-containing small peptides through the degradation of exopolymers was correlated with the reutilization of the tyrosine-like fluorophore. Two humic-like fluorophores that showed emission maxima at the longer wavelength (525 nm) increased during the stationary phase, while the other humic-like fluorophore, which had a shorter emission wavelength (400 nm) and was categorized as recalcitrant, was stable. These humic-like fluorophore behaviors during incubation indicated that the composition of bacterial humic-like fluorophores, which were unavailable to the strain, differed between growth phases. Our results suggest that bacterial physiology can affect RDOM production and accumulation in the ocean interior.

Combined glutathione S-transferase T1 and M1 positive genotypes afford protection against type 2 diabetes in Japanese.

INTRODUCTION: Diabetes mellitus is associated with an increased production of reactive oxygen species and a reduction in antioxidant defenses. The aim of this study is to determine the association between the incidence of Type 2 diabetes and gene polymorphisms of glutathione S-transferase (GST), which modulates oxidative stress. MATERIALS & METHODS: The associations between the incidence of Type 2 diabetes and the GSTT1 and GSTM1 genotypes were analyzed in 469 Japanese participants in a health-screening program. RESULTS: The clinical characteristics and smoking status were obtained from the health screening record. The incidence of diabetes was 1.5-fold higher in the GSTT1 and GSTM1 null (-) genotype than the GSTT1 and GSTM1 present (+) genotype, respectively. Although the effect of each null genotype was not significant, the combined GSTT1+/GSTM1+ genotypes conferred a significant reduction in risk of diabetes in comparison with the other combinations of genotypes (adjusted odds ratio [OR]: 0.30; 95% confidence interval [CI]: 0.12-0.71). In stratified analyses by smoking status, the incidence of diabetes was significantly higher in never-smokers with the GSTT1- genotype than those with the GSTT1+ genotype (OR: 2.85; 95% CI: 1.17-6.94) and increased significantly in current smokers (OR: 5.91; 95% CI: 1.96-17.88). The effect of the GSTM1- genotype was significant only in current smokers. CONCLUSION: This study demonstrated that the GSTT1- and GSTT1-/GSTM1- genotypes are independent risk factors for development of Type 2 diabetes regardless of the smoking status of the patient, and that these genotypes and current smoking were interactively associated with the incidence of Type 2 diabetes.

Population estimation of the effects of cytochrome P450 2C9 and 2C19 polymorphisms on phenobarbital clearance in Japanese.

A nonlinear mixed-effect modeling (NONMEM) program was used to evaluate the effects of cytochrome P450 (CYP) 2C9 and CYP2C19 polymorphisms on the phenobarbital (PB) population clearance for Japanese epileptics. The pharmacokinetics of the 260 PB concentrations at a steady-state obtained from 79 patients was described with a one-compartment open pharmacokinetic model with first-order elimination. The covariates screened included the total body weight (BW), age, gender, PB daily dose, CYP2C9 and CYP2C19 genotypes, the coadministered antiepileptic drugs (AEDs), and complications. The final model of PB apparent clearance was as follows: CL = 0.23 x (BW/40)0.21 x 0.52CYP2C9*1/*3 x 0.68VPA x 0.85PHT x 0.85SMID x (1 + etaCL) where CL = the clearance of PB; CYP2C9*1/*3 = 1, otherwise 0; VPA = 1 if valproic acid is coadministered, otherwise 0; PHT = 1 if phenytoin is coadministered, otherwise 0; SMID = 1 if complications of severe or profound mental retardation with a significant behavior impairment are presented, otherwise 0; and etaCL = the independent random error distributed normally with the mean zero and variance equal to omegaP2. The total clearance of PB decreased by 48% in patients with CYP2C9*1/*3 genotype in comparison with those with CYP2C9*1/*1 genotype (P < 0.001). An effect of CYP2C19 polymorphisms was not detected. To our knowledge, this is the first report to demonstrate that the CYP2C9 genotype affects the PB metabolism in routine care, but the results should be further verified in other ethnic populations.

Association between paraoxonase-1 Q192R polymorphism and lung function among Saskatchewan grain handlers.

INTRODUCTION: Paraoxonase-1 (PON1) is a high-density, lipoprotein-associated, multifunctional antioxidant enzyme that is detected in nonciliated bronchiolar epithelial cells, although its role in the lung has not yet been clarified. We therefore investigated the association between the PON1 Q192R polymorphism and lung function. PATIENTS & METHODS: A total of 216 male Saskatchewan grain handlers provided demographic, occupational and respiratory-symptom information by means of questionnaires, and thereafter underwent PON1 Q192R genotyping and lung-function testing. RESULTS: Mean lung-function values did not differ among the Q192R genotypes. However, current smokers with the Q/Q genotype had a higher mean percent predicted forced expiratory volume in the first second (FEV(1)), and absolute and percent predicted FEV(1) per forced vital capacity (FVC) compared with current smokers with at least one 192R allele (100.9 +/- 11.2% vs 92.0 +/- 15.1%, p = 0.01; 78.0 +/- 5.9% vs 74.1 +/- 6.8%, p = 0.03; and 96.8 +/- 7.1% vs 92.1 +/- 8.3%, p = 0.03; respectively). The incidence of subjects with FEV(1)/FVC less than 70% was significantly higher in current smokers with at least one 192R allele than in nonsmokers with the Q/Q genotype (odds ratio: 5.0; 95% confidence interval: 1.5-17.4). The protective effect of the Q/Q genotype was not found in nonsmokers. The FVC was not influenced by either PON1 genotype or smoking status. CONCLUSION: The results obtained from grain handlers suggest that PON1 may play some role in the protection of the airways against the toxicity of cigarette smoke, and the 192R allele may be a novel genetic risk factor for airway injury.