Alterations in DNA methylation rates of brain-derived neurotrophic factor in patients with schizophrenia

Background and objectives: Schizophrenia (SZ) is one of the most disabling mental illness and the elucidation of diagnostic and therapeutic biomarkers are required. Recent studies investigating the brain morphology, the gene expression profile, and the genetic epidemiology have suggested the involvement of Brain-derived neurotrophic factor (BDNF) and its epigenetic regulation in the pathophysiology of SZ. The current study was conducted to determine the association of DNA methylation of the BDNF gene with the diagnosis or with the characteristics of patients with SZ.MethodsWe analyzed genomic DNA from peripheral blood of 22 patients with SZ and 22 healthy subjects. The DNA methylation rates (DMRs) of the CpG island at the promoter of exón I of the BDNF gene were measured using EpiTYPER® and the MassARRAY® system (Agena Biosciences). We examined the validity of the methylation profiles as a diagnostic biomarker for SZ by clustering analyses, differences in DMRs between patients and healthy controls, and the relationship between DMRs and patient characteristics.ResultsThe clustering analysis failed to distinguish between healthy controls and patients with SZ, though the DMRs of 4 CpG units were significantly different between these two groups. Whereas the DMR of one CpG (CpG 28) was significantly correlated with the amount of daily antipsychotics, there was no influence of age, severity, or duration of illness on the DMRs of the BDNF gene.ConclusionDespite the small number of subjects, our results suggest the involvement of the changes in DMRs of the BDNF gene in the pathophysiology of SZ. (AU)

Arl4c expression in colorectal and lung cancers promotes tumorigenesis and may represent a novel therapeutic target.

We recently demonstrated that expression of ADP-ribosylation factor (ARF)-like 4c (Arl4c) induced by a combination of Wnt/ß-catenin and epidermal growth factor/Ras signaling in normal epithelial cells grown in three-dimensional culture promotes cellular migration and proliferation, resulting in formation of tube-like structures, suggesting the involvement of Arl4c in epithelial morphogenesis. It is conceivable that there could be a common mechanism between epithelial morphogenesis and carcinogenesis. Therefore the current study was conducted to investigate whether Arl4c might be involved in tumorigenesis. Immunohistochemical analyses of tissue specimens obtained from colorectal and lung cancer patients revealed that Arl4c was not observed in non-tumor regions but was strongly expressed at high frequencies in tumor lesions. Inhibition of Wnt/ß-catenin or Ras/mitogen-activated protein kinase signaling reduced Arl4c mRNA levels in HCT116 colorectal cancer cells and A549 lung cancer cells. Knockdown of Arl4c inhibited Rac activity and also prevented nuclear localization of yes-associated protein (YAP)/transcriptional co-activator with PDZ-binding motif (TAZ) in these cancer cells. Arl4c-depleted cancer cells consistently showed decreased migration, invasion and proliferation capabilities both in vitro and in vivo. Furthermore, direct injection of Arl4c small interfering RNA (siRNA) into HCT116 cell-derived tumors (in vivo treatment with siRNA) inhibited tumor growth in immunodeficient mice. These results suggest that Arl4c is involved in tumorigenesis and might represent a novel therapeutic target for suppressing proliferation and invasion of colorectal and lung cancer cells.

[Simultaneous resection of pulmomary metastasis of colon cancer and primary lung cancer].

A 70-year-old woman was admitted with a complaint of weight loss and an abnormal shadow on the chest X-ray. On palpation, the unmobilized mass, measuring 5 cm, was located in the left lower abdomen. The barium-enema examination showed the filling defect of the sigmoid colon. Chest computed tomography (CT) showed a tumor, measuring 20 x 20 mm, located in the right upper lobe (S3) and a nodule, measuring 3 mm, located in the right lower lobe (S8). At first, we performed sigmoidectomy (D 3) for the colon cancer. Next, performed right upper lobectomy and a partial resection of the right lower lobe. Histopathologically, the one is a primary lung cancer, the other is a metastatic lung cancer. With an increase in colorectal and lung cancer, similar cases as ours seem to increase in number. When we treat multiple lung nodules with malignancy of other organs, we should consider 3 types of cases, 1) only primary, 2) primary and metastatic, 3) only metastatic.

Methyl 2-(methylthio)benzoate: the unique sulfur-containing sex pheromone of Phyllophaga crinita.

The female-produced sex pheromone of Phyllophaga crinita (Burmeister) (Coleoptera: Scarabaeidae: Melolonthinae; the adult has no common name) is identified as methyl 2-(methylthio)benzoate. This is the first identification of a sulfur-containing, long-distance, female-produced sex attractant from any insect taxa. The root-feeding larvae of this species are serious pests in many crops in Texas and Mexico. In field tests, many P. crinita males were captured in traps baited with the authentic compound. Interestingly, a heteroatom analog, methyl 2-methoxybenzoate, also captured P. crinita males, but only at a dose 10,000 times higher than the lowest tested dose of the authentic pheromone.

Exon 3 of tyrosine hydroxylase gene: lack of association with Japanese schizophrenic patients.

Tyrosine hydroxylase (TH) is the rate-limiting enzyme in dopamine (DA) biosynthesis. Exon 3 of the human TH gene encodes the sequence from Ser31 to Glu104 of type 1 enzyme, which contains the critical parts for regulation of the catalytic activity. The amino acid residues Gly36-Arg37-Arg38 were identified as a key sequence for DA to exert its inhibitory effect on catalytic activity. Therefore, we screened the nucleotide sequences of exon 3 from 201 Japanese patients with schizophrenia to explain the elevation in the synaptic or presynaptic DA concentrations in the schizophrenic brain, based on the hypothesis that any mutation changing the amino acid sequence Gly36-Arg37-Arg38 would result in the elevation of DA synthesis, due to a reduced inhibitory effect of DA on the catalytic activity. However, no mutated sequences of exon 3 and both exon-intron boundaries were detected in any of the patients examined. Polymorphisms generating Val81 and Met81 were compared of the distributions of genotype and allele between the patients and 175 Japanese healthy controls, which did not suggest an association between the polymorphism and schizophrenia. These results indicate that exon 3 of the human TH gene lacks association with schizophrenia in Japanese patients.

Structure-activity relationship of orally potent tripeptide-based HIV protease inhibitors containing hydroxymethylcarbonyl isostere.

We designed and synthesized a new class of peptidomimetic human immunodeficiency virus protease inhibitors containing a unique unnatural amino acid, allophenylnorstatine [Apns; (2S,3S)-3-amino-2-hydroxy-4-phenylbutyric acid], with a hydroxymethylcarbonyl isostere as the active moiety. From a structure-activity relationship study of HIV-1 protease inhibition, enzyme selectivity for other aspartyl proteases, the antiviral activity and pharmacokinetics in rats, 24c (KNI-227) and 24d (KNI-272, our first clinical candidate) were found to be selective and orally potent HIV protease inhibitors. Moreover, an improvement of the pharmacokinetic features of KNI-272 provided two long-lasting and highly bioavailable compounds (24g: JE-2178, 24h: JE-2179).

The Escherichia coli pldC gene encoding lysophospholipase L(1) is identical to the apeA and tesA genes encoding protease I and thioesterase I, respectively.

We deduced the amino acid sequence of Escherichia coli lysophospholipase L(1) by determining the nucleotide sequence of the pldC gene encoding this enzyme. The translated protein was found to contain 208 amino acid residues with a hydrophobic leader sequence of 26 amino acid residues. The molecular weight of the purified enzyme (20,500) was in good agreement with the predicted size (20,399) of the processed protein. A search involving a data bank showed that the nucleotide sequence of the pldC gene was identical to those of the apeA and tesA genes encoding protease I and thioesterase I, respectively. Consistent with the identity of the pldC gene with these two genes, the enzyme purified from E. coli overexpressing the pldC gene showed both protease I and thioesterase I activities.

Induction of terminal differentiation and apoptosis in human colonic carcinoma cells by brefeldin A, a drug affecting ganglioside biosynthesis.

An appreciable increase in G(M3) with a concomitant decrease in some neolacto-series gangliosides was observed during differentiation of human colonic carcinoma HCT 116 cells induced by a differentiating agent. When the cells were treated with brefeldin A (BFA), a striking increase in de novo biosynthesis of G(M3) and a decrease in biosynthesis of neolactoseries gangliosides were observed after 6 h. Clear morphological changes to differentiated epithelial cells and an arrest of cells in the G0/G1 phase of the cell cycle were observed after 1 day of treatment. Then the cells were led to apoptosis. This activity was not affected by forskolin, which antagonizes the effects of BFA on protein transport and the Golgi apparatus. These results suggest that the differentiation-inducing activity of BFA might be due to its modulatory effect on ganglioside biosynthesis, and that a specific change in ganglioside pattern is an essential prerequisite for induction of differentiation, providing a novel target for differentiation therapy of cancer.

Structure-activity relationship of small-sized HIV protease inhibitors containing allophenylnorstatine.

We designed and synthesized a new class of peptidomimetic human immunodeficiency virus (HIV) protease inhibitors containing a unique unnatural amino acid, allophenylnorstatine [Apns; (2S, 3S)-3-amino-2-hydroxy-4-phenylbutyric acid], with a hydroxymethylcarbonyl (HMC) isostere as the active moiety. A systematic evaluation of structure-activity relationships for HIV protease inhibition, anti-HIV activities, and pharmacokinetic profiles has led to the delineation of a set of structural charateristics that appear to afford an orally available HIV protease inhibitor. Optimum structures, exemplified by 21f (JE-2147), incorporated 3-hydroxy-2-methylbenzoyl groups as the P2 ligand, (R)-5,5-dimethyl-1,3-thiazolidine-4-carbonyl (Dmt) residue at the P1' site, and 2-methylbenzylcarboxamide group as the P2' ligand. The present study demonstrated that JE-2147 has potent antiviral activities in vitro and exhibits good oral bioavailability and plasma pharmacokinetic profiles in two species of laboratory animals.

Changes in composition of newly synthesized sphingolipids of HeLa cells during the cell cycle -- suppression of sphingomyelin and higher-glycosphingolipid synthesis and accumulation of ceramide and glucosylceramide in mitotic cells.

Sphingolipid biosynthesis in synchronized HeLa cells was studied by pulse labeling with [14C]Ser or [14C]Gal and a simple TLC method. The major HeLa cell sphingolipids are ceramide (Cer), sphingomyelin, glucosylceramide (GlcCer), lactosylceramide (LacCer), globotriaosylceramide (Gb3Cer), N-acetylneuraminosylgangl iotriaosylceramide (GM2) and sialylparagloboside (G[M1-GlcNAc]). The sphingolipid biosynthetic profiles of HeLa cells in the G1, G1/S boundary, S and G2 phases were similar, but significant changes occurred during M phase, when incorporation of radioactivity into sphingomyelin, Gb3Cer and a mixture of GM2 and G(M1-GlcNAc) decreased, and those of Cer and GlcCer increased. These data indicate that transfer of phosphocholine and galactose to Cer and GlcCer, respectively, decreased in mitotic cells, resulting in accumulation of Cer and GlcCer. Analysis of LacCer synthase activity revealed that GlcCer accumulation was not due to reduced activity of this enzyme. The results suggest that Cer and GlcCer accumulation in mitotic cells resulted from suppression of sphingomyelin and LacCer synthesis, probably caused by vesiculation of membranous organelles, such as the endoplasmic reticulum and Golgi apparatus.

Activated mast cells release extracellular type platelet-activating factor acetylhydrolase that contributes to autocrine inactivation of platelet-activating factor.

IgE-dependent and -independent activation of mouse bone marrow-derived mast cells (BMMC) elicited rapid and transient production of platelet-activating factor (PAF), which reached a maximal level by 2-5 min and was then degraded rapidly, returning to base-line levels by 10-20 min. Inactivation of PAF was preceded by the release of PAF acetylhydrolase (PAF-AH) activity, which reached a plateau by 3-5 min and paralleled the release of beta-hexosaminidase, a marker of mast cell exocytosis. Immunochemical and molecular biological studies revealed that the PAF-AH released from activated mast cells was identical to the plasma-type isoform. In support of the autocrine action of exocytosed PAF-AH, adding exogenous recombinant plasma-type PAF-AH markedly reduced PAF accumulation in activated BMMC. Furthermore, culture of BMMC with a combination of c-kit ligand, interleukin-1beta and interleukin-10 for > 24 h led to an increase in plasma-type PAF-AH expression, accompanied by a reduction in stimulus-initiated PAF production. Collectively, these results suggest that plasma-type PAF-AH released from activated mast cells sequesters proinflammatory PAF produced by these cells, thereby revealing an intriguing anti-inflammatory aspect of mast cells.

Separation of 18 6-aminoquinolyl-carbamyl-amino acids by ion-pair chromatography.

Eighteen 6-aminoquinolyl-carbamyl (AQC)-amino acids were separated by ion-pair chromatography, using tetrabutylammonium (TBA) as a counterion. Optimum separation was obtained on a C8 reverse-phase column using gradient elution with two mobile phases, A (5 mM TBA, 75 mM ammonium acetate, pH 7.5) and B (80% acetonitrile). The AQC-amino acids were detected by fluorescence with excitation at 250 nm and emission at 395 nm, and the analysis time was 65 min. The response factors of individual AQC-amino acids to AQC-phenylalanine ranged from 0.42 to 1.08 (except for tryptophan at 0.01), with an average of 0.8. Detection limits by fluorescence ranged from 11.8 fmol (threonine) to 51.7 fmol (methionine), except for tryptophan (1.8 pmol).

[Decision-making style among hemodialysis patients].

UNLABELLED: The purpose of this descriptive study was to identify the decision-making style among the dialysis patients. Subjects were a Convenience sample of hemodialysis patients who agreed to participate this study. The data was collected through semi-structured interviews. The interviews were recorded on the tape and transcribed. Decision-making episodes were picked and interpreted by searchers. Data was analyzed based on the Inductive Theory Approach. RESULT: Subject were 21 patients from 4 hospitals. The mean age was 51.1 and the mean duration after they started the hemodialysis was 9.1 years. The 16 decision-making styles were identified: accept, unvacillate, resignation, compromise, perseverance, challenge, self-activation, groupie, crisis-avoidance, withdrawal, prudent, reserve, escape, refusal, dogmatic, and entrust. These 16 decision-making styles were characterized by the reality oriented perception, the constructive regimes, the higher quality of social support. On the other hand, the passive decision-making styles were characterized by the distorted perception, the passive regimes, and the poor social support. DISCUSSION: The reality orientation, energy, openness, values system were discussed as key factors which determined the decision-making style among hemodialysis patients. The nursing approach supporting the patients to make their decision were discussed.

[Components of decision-making among hemodialysis patients].

UNLABELLED: The purpose of this descriptive study was to identify the components of decision-making among the dialysis patients. Subjects were a Convenience sample of hemodialysis patients who agreed to participate this study. The data was collected through semi-structured interviews was recorded on the tape and transcribed. Decision-making episodes were picked and interpreted by researchers. Data was analysed based on the Grounded Theory Approach. RESULT: Subject were 21 patients from 4 hospitals. The mean age was 51.1 and the mean duration after the hemodialysis was 9.1 years. The number of episode of decision-making were 136. Six components of decision-making were extracted such as . The subject were making decision by examining and judging the situation and oneself, and then setting the goal. They were selecting situation and themselves. After all, the outcomes of decision-making were evaluating. Seven type of decision-making, erection type, reality-oriented type, elaborate type, dogmatic type, passive type, chance type, resistant type were extracted. DISCUSSION: The factors which were related to the quality of decision-making were discussed such as the ability to examine the reality and control own wishes and needs, and the commitment for the decision-making. The nursing approach to facilitate the patients to make the decision were mentioned.

Cloning, expression and characterization of plasma platelet-activating factor-acetylhydrolase from guinea pig.

In a previous study, we purified PAF-acetylhydrolase, which converts PAF to an inactive metabolite, lysoPAF, from peritoneal fluid of guinea pigs subjected to experimental endotoxin shock and found that this purified enzyme had similar biochemical properties to the plasma enzyme [Karasawa, K., Yato, M., Setaka, M., and Nojima, S. (1994) J. Biochem. 116, 374-379]. In this study, we isolated a homogeneous enzyme preparation from guinea pig plasma using a similar procedure. The molecular mass of this purified enzyme, as determined by SDS-PAGE was 58-63 kDa, larger than that (43 kDa) of the human enzyme. To elucidate the molecular structure of this enzyme and clarify its relationships with PAF-acetylhydrolases of other species, we isolated and sequenced a cDNA encoding this enzyme. Its cDNA contains an open reading frame encoding 436 amino acids and its predicted molecular mass (49 kDa) is lower than that of the native enzyme, suggesting that guinea pig plasma PAF-acetylhydrolase, unlike the human enzyme, is modified post-translationally, perhaps by glycosylation.

A distribution study of 11C platelet-activating factor (PAF) analogs in normal and tumor-bearing mice.

As a preliminary study to image platelet-activating factor (PAF) receptors in vivo, comparative study of biodistribution between 1-O-hexadecy1-2-O-N, N-dimethylcarbamoyl-sn-glycero-3-phosphocholine [choline-methyl-11C](L-[11C]dimethylcarbamoyl-PAF) and nonspecific PAF analog, 3-O-hexadecyl-2-O-N,N-dimethylcarbamoyl-sn-glycero-1-phosphocholine [choline-methyl-11C](D-[11C]-dimethylcarbamoyl-PAF) was carried out in both normal and tumor-bearing mice. Higher accumulation of L-[11C]dimethylcarbamoyl-PAF than D-[11C]dimethylcarbamoyl-PAF was observed in normal mice spleen. The co-administration of PAF antagonists dose-dependently reduced the radioactivity level of the L-isomer only in the spleen. In mice bearing Ehrlich tumors and Sarcoma 180, more L-than the D-[11C]-isomer was accumulated in the tumor and spleen. We found that specific accumulation sites for L-[11C]dimethylcarbamoyl-PAF exist in the spleen and tumors than in other tissues. Moreover, the comparison of accumulation between L- and D-[11C] dimethylcarbamoyl-PAF would be a useful procedure for estimation of PAF receptors in vivo.

[Characteristics of psychosocial nursing care--the dimensions of psychosocial nursing care to promote empowerment among patients].

UNLABELLED: Nurses provide the psychosocial nursing care to promote the patients' empowerment. In spite of recognizing the importance of psychosocial nursing. The research reported here is part of a large study designed to explore the psychosocial nursing intervention strategies. This part of the study is focused on characteristics of psychosocial nursing care as identified by nurses. Research design was descriptive and qualitative. Subjects were a convenient sample of nurses who have more than 5 years experience and agreed to participate in this research. Data was collected through semistructured interviews. Data was recorded on a tape and transcribed, then analyzed based on Grounded Theory Approach. SUBJECTS: The number of subjects consisted of 39 nurses. The average age and duration of clinical experience was 36.6 years old and 12.6 years. RESULT: Sixteen of the characteristics of psychosocial nursing to promote the empowerment among the patients were identified; 1) respect the patients' decision making, 2) clinical judgment from multiple perspectives, 3) predictability, 4) flexibility, 5) classification of the patients by comparison, 6) monitoring, 7) relief from reality, 8) relief from pain and suffering, 9) enhancement of the patients' ego strength, 10) nurture of hope, 11) support individualized daily living styles, 12) encouragement of the patients'own problem solving abilities, 13) respect of individuality, 14) self-utilization, 15) self-control based on professional identity, 16) utilization of team power. DISCUSSION: Nurses choose from the sixteen dimensions of psychosocial nursing and organizing them in order to stabilizing the patients' conditions.