Genetic Association Analysis of NOS3 and Methamphetamine-Induced Psychosis Among Japanese.

Endothelial nitric oxide synthase (NOS3) is one of the enzymes influencing nitric oxide (NO) function in the human brain. NO is a gaseous neurotransmitter that is involved in a variety of mechanisms in the central nervous system, such as N-methyl-D-aspartate receptor activation and oxidative stress. The evidence from animal pharmacological studies and postmortem studies supports an association between NO and psychotic disorders. Methamphetamine (METH) use disorder is a known psychotic disorder, and we therefore conducted a gene-based case-control study between tagging single nucleotide polymorphisms (SNPs) (rs2070744, rs1799983) in NOS3 and METH-induced psychosis in Japanese subjects (183 with METH-induced psychosis and 267 controls). Written informed consent was obtained from each subject. No significant association was found between any tagging SNP in NOS3 and METH-induced psychosis in the allele/genotype-wise or haplotype-wise analyses. In conclusion, we suggest that NOS3 might not contribute to the risk of METH-induced psychosis in the Japanese population.

SIRT1 gene, schizophrenia and bipolar disorder in the Japanese population: an association study.

Several lines of evidence suggest that alterations in circadian rhythms might be associated with the pathophysiology of psychiatric disorders such as schizophrenia and bipolar disorder (BP). A recent study reported that SIRT1 is a molecule that plays an important role in the circadian clock system. Therefore, to evaluate the association among the SIRT1 gene, schizophrenia and BP, we conducted a case-control study of Japanese population samples (1158 schizophrenia patients, 1008 BP patients and 2127 controls) with four tagging SNPs (rs12778366, rs2273773, rs4746720 and rs10997875) in the SIRT1 gene. Marker-trait association analysis was used to evaluate the allele and the genotype association with the χ(2) test, and haplotype association analysis was evaluated with a likelihood ratio test. We showed an association between rs4746720 in the SIRT1 gene and schizophrenia in the allele and the genotype analysis. However, the significance of these associations did not survive after Bonferroni's correction for multiple testing. On the other hand, the SIRT1 gene was associated with Japanese schizophrenia in a haplotype-wise analysis (global P(all markers) = 4.89 × 10(-15)). Also, four tagging SNPs in the SIRT1 gene were not associated with BP. In conclusion, the SIRT1 gene may play an important role in the pathophysiology of schizophrenia in the Japanese population.

Clinical features associated with the homozygous Trp64Arg mutation of the beta3-adrenergic receptor: no evidence for its association with obesity in Japanese.

To characterize the clinical features associated with the Trp64Arg mutation of the beta3-adrenergic receptor (beta3-AR), the effects of this mutation, in particular the homozygous state (Arg/Arg), on obesity, blood pressure, and plasma lipoproteins were investigated in 2 populations: subjects residing on a small isolated island (group 1; n=746) and patients residing in Tokyo who attend a clinic for metabolic diseases (group 2; n=371). The allelic frequency of the Trp64Arg mutation was 23.4% in group 1 and 18.3% in group 2. No significant difference in the body mass index was observed between subjects with 3 different genotypes in each group. There was a trend that the Arg/Arg had higher systolic blood pressure than the Trp/Trp in both groups, but the differences were not statistically significant. The plasma LDL cholesterol levels were significantly lower in Arg/Arg than in Trp/Trp in men from the group 1 cohort (2.82+/-0.84 versus 3.19+/-0.7 mmol/L, P<0.05). These results suggest that the homozygous Trp64Arg mutation is not a major contributing factor for obesity, but potentially contributed to higher systolic blood pressure and low plasma levels of LDL cholesterol in Japanese men.

Cholesterol-lowering effects of psyllium seed associated with urea metabolism.

Twenty-eight mild hypercholesterolemic male and female adults were orally administered psyllium seed for 3 months. After psyllium treatment, the serum total cholesterol, low-density-lipoprotein-cholesterol and atherogenic index significantly decreased, but levels of high-density-lipoprotein-cholesterol, triglyceride and urea nitrogen did not. To determine the parameters associated with the cholesterol-lowering effect in the subjects' backgrounds, both biochemical and hematological parameters, we statistically examined the correlation between pretreatment parameters and the absolute change of total cholesterol level. The absolute change of total cholesterol level showed a direct correlation with the triglyceride level at pretreatment (r=0.41, P=0.03) and had an inverse correlation with urea nitrogen level (r=-0.46, P=0.01) but not with the total cholesterol level (r=-0.18). The change in urea nitrogen level had an inverse correlation with the urea nitrogen level itself at pretreatment (r=-0.82, P=7 x 10[-8]) and had a direct correlation with the triglyceride level (r=0.43, P=0.02). The change in triglyceride level had an inverse correlation with the urea nitrogen level (r=-0.48, P=0.008). Furthermore, the change in total cholesterol level had direct correlations with changes in the triglyceride level (r=0.56, P=0.002) and the urea nitrogen level (r=0.51, P=0.006), but these changes in triglyceride and urea nitrogen level did not correlate significantly. These findings suggest the close association of urea nitrogen and lipid metabolism in hyperlipidemia and psyllium seed treatment.

Acute comitant esotropia in a boy with head trauma and convulsions receiving carbamazepine.

We examined an 11-year-old boy who complained of acute onset of diplopia. The patient had head trauma and postsurgical convulsions and had been treated with carbamazepine. Diplopia developed after the dose of carbamazepine was increased to 700 mg/day. On examination, comitant esotropia and lateral gaze nystagmus were found. These disorders disappeared after carbamazepine was decreased to 400 mg/day. We believe that acute comitant esotropia and lateral gaze nystagmus may have been precipitated by head trauma and carbamazepine in our patient.

Influences of supplementary dietary tungsten on methionine metabolism in rabbits fed a low-cholesterol plus methionine diet.

Hyperhomocysteinemia results from an impaired methionine metabolism. Sulfite oxidase, which is an important enzyme in methionine metabolism, contains molybdenum. In contrast, tungsten has a molybdenum-antagonistic effect. Thus, we hypothesized that dietary tungsten may decrease plasma homocysteine levels and influence methionine metabolism. Male New Zealand White rabbits (n=15) were fed a low-cholesterol basal diet and then placed on three different diets: 0.1% cholesterol (Chol), Chol plus 1% methionine (Met), and Chol plus Met plus 0.1% tungsten (W). The animals received these diets for 20 weeks. Biochemical tests of blood and urine were performed. Plasma homocysteine levels were significantly lower in the Chol+Met+W group than in the Chol+Met group. Plasma levels of total cholesterol, triglyceride, lipid peroxide, and urinary 24-h taurine concentrations were higher in the Chol + Met + W group than in the Chol + Met group. In comparison, concentrations of 2, 3-diphosphoglycerate (2, 3-DPG), reduced glutathione (GSH) in erythrocytes, and urinary 24-h SO4(2) were lower in the Chol+Met+W group than in the Chol+Met group. From these results, tungsten could be expected to exhibit an antiatherogenic effect. Conversely, it may have effects on atherogenic factors. Thus, tungsten may play a number of roles in the methionine metabolism.

[Changes in serum Lp(a) and apo(a)phenotype in patients with acute cerebral stroke].

33 patients with acute cerebral stroke (21 men and 12 women, averaged age 69.9 +/- 11.2 years) were examined for fluctuations in the level of serum Lp(a) 3, 7, 14, and 28 days after the onset of symptoms. The Lp(a) level was higher (p < 0.05, p < 0.001) on days 7 and 14 than on the day of onset. Higher values were associated with poorer outcomes and with larger abnormal densities on brain CT scans. In 33 of these patients, the apo(a)phenotype was determined. Diluted serum was electrophoresed with SDS-containing gel as a carrier, then transcribed on a nitrocellulose membrane, stained by Western blotting and then sensitizing-stained. The results were evaluated by the method of Utermann et al. Outcome was significantly poorer in patients with a double-band apo(a)phenotype than in those with a single-band (p < 0.05). Those with a double-band had significantly higher mean value of Lp(a), except on the day of onset, than did those with a single-band. Patients with large degrees of electrophoresis had significantly higher values of Lp(a) than did those with small degrees. These data suggest that serial measurement of Lp(a) can be of some value in predicting the outcome in patients with cerebral stroke. Determination of apo(a) phenotype may also be useful in the prevention and prognosis of cerebral stroke, especially when it is caused by cerebral infarction.

A lipid lowering drug (bezafibrate) has a favorable effect on liver enzymes (Al-P and gamma-GTP).

We administered 400 mg of bezafibrate daily to 27 patients with hyperlipidemia for seven moths. Most biochemical parameters remained unchanged, whereas levels of alkaline phosphatase (Al-P)and gamma-glutamyl transpeptidase (gamma-GTP), which are the hepatobiliary enzymes, were significantly decreased. Blood lipid levels were improved. Al-P levels decreased significantly from the baseline level of 174.5 IU/l to 116.7 IU/l (-26.5%) and gamma-GTP levels also decreased from 64.4 IU/l to 34.4 IU/l (-29.5%) (p < 0.001). When we compared the changes in serum lipid levels with those in Al-P and gamma-GTP levels following bezafibrate therapy, we found a slight degree of correlation between changes in gamma-GTP and triglyceride (T-G) levels, but no correlation at all between the changes of Al-P and total-cholesterol (T-cho), T-G or high density lipoprotein cholesterol (HDL-C). A close correlation was observed between both Al-P and gamma-GTP (r = 0.81, p < 0.001). From these results it was suggested that bezafibrate has not only a lipid lowering effect but has a favorable efficacy on the hepatobiliary enzymes.

Adult-onset spinocerebellar dysfunction caused by a mutation in the gene for the alpha-tocopherol-transfer protein.

BACKGROUND: Patients with isolated vitamin E deficiency have an impaired ability to incorporate alpha-tocopherol into lipoproteins in the liver and usually have symptoms and signs of spinocerebellar dysfunction before adolescence. Accumulated evidence suggests that the alpha-tocopherol-transfer protein, which is presumed to function in the intracellular transport of alpha-tocopherol, is abnormal in these patients. METHODS: We studied a patient from an isolated Japanese island who began to have ataxia, dysarthria, and sensory disturbances in the sixth decade of life. His serum vitamin E concentration was low (1.2 micrograms per milliliter [2.8 mumol per liter]). Exons of his gene for the alpha-tocopherol-transfer protein were analyzed by DNA sequencing. We also screened an additional 801 inhabitants of the island for the mutation. Both the normal and mutant alpha-to-copherol-transfer proteins were expressed in COS-7 cells and studied by immunoblot analysis and assay for alpha-tocopherol-transfer activity. RESULTS: The patient was homozygous for a point mutation that replaces histidine (CAT) with glutamine (CAG) at position 101 of the gene for the alpha-tocopherol-transfer protein. When expressed in COS-7 cells, the missense mutation produced a functionally defective alpha-tocopherol-transfer protein with approximately 11 percent of the transfer activity of the wild-type protein. Of the 801 island inhabitants examined, 21 were heterozygous for the His101Gln mutation. In all affected subjects, including the patient, this mutation cosegregated with an intron-sequence polymorphism. The heterozygotes were phenotypically normal and had serum vitamin E concentrations that were on average 25 percent lower than those of normal subjects (mean [+/- SD], 7.5 +/- 2.2 vs. 10.1 +/- 2.8 micrograms per milliliter [17.4 +/- 5.1 vs. 23.4 +/- 6.5 mumol per liter]; P = 0.002). CONCLUSIONS: alpha-Tocopherol-transfer protein is a determinant of serum vitamin E concentrations. An abnormality in this protein is a cause of spinocerebellar dysfunction.

Effects of an HMG-CoA reductase inhibitor on cytokine production by human monocytes/macrophages.

It has been reported that the HMG-CoA reductase inhibitor simvastatin does not always effectively lower plasma LDL. This drug acts to monocytes/macrophages directly and inhibits cholesterol ester accumulation in these cells. However cytokine production in macrophages when simvastatin was administrated has not been described. In this study, we examined whether simvastatin affects cytokine production in human monocyte-derived macrophages. Simvastatin at doses ranging from 10(-9) to 10(-5) M did not affect the synthesis of proinflammatory cytokines (IL-1 beta, IL-6, IL-8) from human peripheral mononuclear cells. In addition, any changes in cytokine-induced cytokine production (IL-1-induced IL-8 synthesis) were not detected after the addition of simvastatin. The present results suggest that simvastatin suppresses foam cell formation in monocyte/macrophage, without affecting the immunological or inflammatory functions of these cells.

Cross-species transplantation of photoreceptors with tacrolimus hydrate (FK506) treatment.

Photoreceptors from normal neonatal or transgenic neonatal mice with normally developing retinas were transplanted into the subretinal spaces of normal rats. The animals with transplants received either immunosuppressive treatment with tacrolimus hydrate for the first two weeks after transplantation or no treatment. At 4 weeks after transplantation, graft survival occurred only in rats treated with tacrolimus. In 7 of the 12 treated eyes, photoreceptors survived at least 4 weeks after transplantation. Relatively mature outer segments were found only in 2 eyes in which the retinal grafts formed rosettes.

Nondetectable cone and rod electroretinographic responses in a patient with Cockayne syndrome.

A 10-year-old girl complained or poor vision in both eyes. The patient showed progeria, physical and mental retardation, sensorineural hearing loss, cutaneous photosensitivity, hyperopia, poor pupillary dilation, exotropia, salt-and-pepper fundi, nondetectable cone and rod electroretinographic (ERG) responses, cerebral atrophy on computed tomography, and demyelination of periventricular white matter on magnetic resonance imaging. We believe that nondetectable cone and rod ERG responses in Cockayne syndrome, as demonstrated in our patient, may be uncommon.

[Effect of tumor necrosis factor and interferon-gamma on platelet-derived growth factor (PDGF) A and B gene expression in THP-1 monocytic cells].

Platelet-derived growth factor (PDGF) is a potent mitogen of mesenchymal cells. A and B chains of PDGF are transcribed in atherosclerotic plaques but details of the mechanism of the expression of either chains in vivo are not understood. It has been suggested that differentiated macrophages, which are the major origin of foam cells, but not monocytes hardly secrete PDGF in vitro. In the present study, we investigated the effect of 1) tumor necrosis factor-alpha (TNF-alpha), a macrophage derived cytokine which promote the differentiation from monocytes to macrophages, and 2) interferon-gamma (IFN-gamma), a T cell derived macrophage activating factor, on the production and mRNA expression of PDGF A and B chains in a human monocytic leukemia cell line THP-1. THP-1 cells were cultured in the presence of one of the experimental cytokines with or without the addition of phorbol 12-myristate 13-acetate (PMA). PMA is known to induce the differentiation of monocytes (including THP-1 cells) to macrophages within a period of 24 hours. Thus the treatment of THP-1 cells with a cytokine and/or PMA in vitro was expected to mimic the effect of the cytokine during the differentiation of monocytes to macrophages in vivo. Unstimulated THP-1 cells expressed no detectable amount of PDGF A or B mRNA. The treatment of THP-1 cells with PMA resulted in the strong expression of both PDGF A and B mRNAs. Stimulated with TNF-alpha (50 U/ml) on day 0, THP-1 cells expressed PDGF A mRNA significantly from 1 up to 3 days after the treatment, whereas the expression of PDGF B and mRNA was only weakly detected on day 2. The maximum signal of PDGF A mRNA was ten times stronger than that of PDGF B mRNA. Within the same period, relatively small number of TNF-alpha treated THP-1 cells had differentiated to macrophages as detected by the reduction of nitro blue tetrazolium (NBT) (35% of total cells) and by the counting of adherent cells (35% of total cells). THP-1 cells simultaneously treated with TNF-alpha and PMA strongly expressed PDGF A and B gene. Most remarkably, the expression of PDGF B mRNA was greatly enhanced with compared to the treatment with PMA alone. In addition, TNF-alpha augmented the secretion of PDGF proteins in PMA stimulated THP-1 cells. In contrast, INF gamma per se had no effect on PDGF gene expression, NBT reducing activity and cell adherence in unstimulated THP-1 cells.(ABSTRACT TRUNCATED AT 400 WORDS)

[The effects of transcorneal administration of prostaglandin E2 on rabbit eyes].

The pharmacologic effects of prostaglandin E2 (PGE2) on the anterior ocular segment were investigated. PGE2 was administered with a glass cylinder attached to the cornea in concentrations ranging from 0.05 to 500 micrograms/ml. The intraocular inflammatory reaction was assessed according to the changes in the anterior chamber flare, ocular tension, and components of the aqueous humor. We also performed experiments designed to inhibit the reactions induced by PGE2 using anti-inflammatory agents. The PGE2 elicited a single peak reaction in the anterior chamber flare, which recovered after several hours, and a transient elevation in the ocular tension in the early stage after PGE2 administration. Quantitative analysis of these two reactions revealed that both were dependent on the dose of PGE2 administered. The concentrations of ascorbic acid and glutathione, particularly that of the reduced form of the latter, were transiently reduced in the aqueous humor. The anterior chamber flares were suppressed by agents that inhibit arachdonic acid metabolism. These findings suggest that the metabolism of arachdonic acid becomes newly activated as a result of stimulation by PGE2 of extrinsic origin.

[The significance of determination of Lp(a) in health examinations].

Lp(a), an independent risk factor of thrombotic and arteriosclerotic diseases, was determined in subjects undergoing health examinations, and the significance of the determination of Lp(a) in such examinations was investigated by studying its relation wih other risk factors for arteriosclerosis, etc. The subjects were 838 individuals. Lp(a) was determined by latex immunoassay (LIA). The mean Lp(a) value for all of the subjects was 10.9 +/- 7.2 mg/dl. Both gender groups were compared by age, but no significant changes were observed. In a study of Lp(a) in accordance with complications, there was no significant difference between the DM group and the non-DM group. There was also no significant difference between the IHD group and the non-IHD group. In the hyperlipemia group, the value of Lp(a) tender to be higher than in the non-hyperlipemia group. In the abnormal ECG group, the Lp(a) value was significantly higher than in the normal ECG group. When the relation between Lp(a) and other factors was studied, there was positive correlation with TC, beta Lp and LDLC, and a significant negative correlation with TRG. There was significant negative correlation with GOT, GPT and TTT. When the incidence of disease was compared by cut-off value, the incidence of abnormal ECGs was significantly higher at Lp(a) values of 25 mgdl or higher. In this study, Lp(a) showed positive correlations with TC, beta Lp and LDLC, the atherogenic risk of Lp(a) was evident. Because of the significant incidence of abnormal ECGs at the Lp(a) cut-off value of 25 mg/dl or higher, the risk range for Lp(a) should probably be considered as 25 mg/dl or higher.(ABSTRACT TRUNCATED AT 250 WORDS)