Novel formula using triceps skinfold thickness to revise the Cockcroft-Gault equation for estimating renal function in Japanese bedridden elderly patients

In recumbent elderly patients, creatinine clearance (eCCr) estimated by the Cockcroft-Gault (CG) equation may not necessarily reflect renal function. We aimed to develop a novel formula to revise the CG equation using anthropometric measurements in bedridden elderly patients and evaluate its clinical utility. The subjects included 77 bedridden Japanese patients aged ≦ 65, hospitalized at Naruto Yamakami Hospital. The actual CCr (mCCr) value was measured using the 24-hour urine collection method. Anthropometric data, such as skeletal muscle mass, body fat mass (BFM), and triceps skinfold thickness (TSF), were collected. We established a novel formula to estimate CCr or CCr by correcting the eCCr value with BFM or TSF. The stage of classification of renal dysfunctions in patients with eGFR or eGFR was equivalent to the GFR based on the mCCr. Notably, the novel equation for eCCr based on TSF (eCCr), dubbed the "Naruto" formula, can be useful to evaluate renal function in bedridden elderly patients without expensive equipment or additional costs. In this study, mCCr was considered to be the true renal function of the patient, but whether and to what extent mCCr correlates with inulin clearance is unknown. J. Med. Invest. 65:195-202, August, 2018.

The Journal of Medical Investigation : JMI


What is the correct adjustment protocol for serum creatinine value to reflect renal function in bedridden elderly patients?

The Japanese Journal of Nephrology and Pharmacotherapy


Genome-wide association study identifies ERBB4 on 2q34 as a novel locus associated with sperm motility in Japanese men.

The decrease in sperm motility has a potent influence on fertilisation. Sperm motility, represented as the percentage of motile sperm in ejaculated sperms, is influenced by lifestyle habits or environmental factors and by inherited factors. However, genetic factors contributing to individual differences in sperm motility remain unclear. To identify genetic factors that influence human sperm motility, we performed a genome-wide association study (GWAS) of sperm motility. A two-stage GWAS was conducted using 811 Japanese men in a discovery stage, followed by a replication study using an additional 779 Japanese men. In the two-staged GWAS, a single nucleotide polymorphism rs3791686 in the intron of gene for erb-b2 receptor tyrosine kinase 4 () on chromosome 2q34 was identified as a novel locus for sperm motility, as evident from the discovery and replication results using meta-analysis (β=-4.01, combined P=5.40×10). Together with the previous evidence that Sertoli cell-specific -knockout mice display an impaired ability to produce motile sperm, this finding provides the first genetic evidence for further investigation of the genome-wide significant association at the locus in larger studies across diverse human populations.


Association of TUSC1 and DPF3 gene polymorphisms with male infertility

Recently, genome-wide association studies of a Hutterite population in the USA revealed that five single nucleotide polymorphisms (SNPs) with a significant association with sperm quality and/or function in ethnically diverse men from Chicago were significantly correlated with family size. Of these, three SNPs (rs7867029, rs7174015, and rs12870438) were found to be significantly associated with the risk of azoospermia and/or oligozoospermia in a Japanese population. In this study, we investigated whether the rs10966811 (located in an intergenic region between the TUSC1 and IZUMO3 genes) and rs10129954 (located in the DPF3 gene) SNPs, previously related to family size, are associated with male infertility. In addition, we performed association analysis between rs12348 in TUSC1 and rs2772579 in IZUMO3 and male infertility. We genotyped 145 patients with infertility (including 83 patients with azoospermia and 62 with oligozoospermia) and 713 fertile controls by PCR-RFLP technique for polymorphism. Because rs10966811 has no restriction sites, the SNP rs12376894 with strong linkage disequilibrium was selected as an alternative to rs10966811. There was a statistically significant association between rs12376894 proxy SNP of rs10966811 and oligozoospermia. Also, a statistically significant association between rs10129954 and azoospermia, and oligozoospermia was observed. When we assessed the relationship between rs12348 in TUSC1 and rs2772579 in IZUMO3 and male infertility traits, we found that rs12348 in TUSC1 was significantly associated with azoospermia and oligozoospermia, but rs2772579 in IZUMO3 was not associated with male infertility. We found that the polymorphisms in TUSC1 and DPF3 displayed strong associations with male infertility.

Journal of Assisted Reproduction and Genetics


An independent validation study of three single nucleotide polymorphisms at the sex hormone-binding globulin locus for testosterone levels identified by genome-wide association studies


Protein kinase C-δ signaling regulates glucagon secretion from pancreatic islets

Accumulating evidence supports the "glucagonocentric hypothesis", in which antecedent -cell failure and inhibition of glucagon secretion are responsible for diabetes progression. Protein kinase C (PKC) is involved in glucagon secretion from -cells, although which PKC isozyme is involved and the mechanism underlying this PKC-regulated glucagon secretion remains unknown. Here, the involvement of PKC in the onset and progression of diabetes was elucidated. Immunofluorescence studies revealed that PKC was expressed and activated in -cells of STZ-induced diabetic model mice. Phorbol 12-myristate 13-acetate (PMA) stimulation significantly augmented glucagon secretion from isolated islets. Pre-treatment with quercetin and rottlerin, PKC signaling inhibitors, significantly suppressed the PMA-induced elevation of glucagon secretion. While Go6976, a Ca(2+)-dependent PKC selective inhibitor did not suppress glucagon secretion. Quercetin suppressed PMA-induced phosphorylation of Tyr(311) of PKC in isolated islets. However, quercetin itself had no effect on either glucagon secretion or glucagon mRNA expression. Our data suggest that PKC signaling inhibitors suppressed glucagon secretion. Elucidation of detailed signaling pathways causing PKC activation in the onset and progression of diabetes followed by the augmentation of glucagon secretion could lead to the identification of novel therapeutic target molecules and the development of novel therapeutic drugs for diabetes. J. Med. Invest. 64: 122-128, February, 2017.


A replication study of a candidate locus for follicle-stimulating hormone levels and association analysis for semen quality traits in Japanese men

In men, follicle-stimulating hormone (FSH) acts on the seminiferous tubules and enhances spermatogenesis. Recently, a candidate locus (rs2414095) for FSH levels was identified by a genome-wide association study (GWAS) in Chinese men. The rs2414095 single-nucleotide polymorphism (SNP) is found on the third intron of the cytochrome P450, family 19, subfamily A, peptide 1 (CYP19A1) gene encoding an aromatase. In the present study, we performed a replication study in 1687 Japanese men (901 from cohort 1 and 786 from cohort 2) to assess whether this SNP is associated with circulating FSH levels. Furthermore, we investigated whether the rs2414095 SNP is correlated with semen quality traits in 2015 Japanese men (1224 from cohort 1 and 791 from cohort 2). The rs2414095 SNP was significantly associated with circulating FSH levels (STD=0.15, P=9.7 × 10(-5)), sperm concentration (STD=0.073, P=0.032) and total sperm number (TSN) (STD=0.074, P=0.027) in a combined analysis of the two Japanese male cohorts. We successfully replicated, in Japanese men, the results of the previous GWAS for the rs2414095 SNP in Chinese men, and found that the rs2414095 SNP was related with sperm production.


Labelling of endogenous target protein via N-S acyl transfer-mediated activation of N-sulfanylethylanilide


Lack of replication of four candidate SNPs implicated in human male fertility traits: a large-scale population-based study

Are the four candidate loci (rs7867029, rs12870438, rs7174015 and rs724078) for human male fertility traits, identified in a genome-wide association study (GWAS) of a Hutterite population in the USA, associated with semen quality traits in a Japanese population? The four single nucleotide polymorphisms (SNPs) rs7867029, rs12870438, rs7174015 and rs724078 have no association with semen parameters in a meta-analysis of two Japanese male cohorts. Four (rs7867029, rs12870438, rs7174015 and rs724078) of the SNPs associated with family size or birth rate in the GWAS of a Hutterite population in the USA were associated with semen parameters in ethnically diverse men from Chicago, USA. This is a replication study in a total of 2015 Japanese subjects, including 791 fertile men and 1224 young men from the general population. We performed a replication study in two cohorts to assess whether the SNPs rs7867029, rs12870438, rs7174015 and rs724078 are associated with sperm concentration, semen volume, total sperm numbers, total motile sperm numbers or sperm motility. The rs12870438 SNP was detected by restriction fragment length polymorphism PCR while rs7174015, rs724078 and rs7867029 SNPs were genotyped using TaqMan probes. This study indicated that none of the four SNPs rs7867029, rs12870438, rs7174015 and rs724078 displayed a significant association with semen parameters in the meta-analysis of two Japanese male cohorts. Only four SNPs identified in the Hutterite GWAS were examined for associations with semen quality traits in a Japanese population. In addition, the linkage disequilibrium structures around the testing markers were different between ethnic groups. Locus mapping studies using a set of tagging SNPs across the loci will be necessary in populations with larger sample sizes in order to understand the contribution of specific genes to semen quality. This study was supported in part by the Ministry of Health and Welfare of Japan (1013201) (to T.I.), Grant-in-Aids for Scientific Research (C) (23510242) (to A.Ta.) from the Japan Society for the Promotion of Science, the European Union (BMH4-CT96-0314) (to T.I.), and the Takeda Science Foundation (to A.Ta.). None of the authors has any competing interests to declare.


An association study of four candidate loci for human male fertility traits with male infertility

Are the four candidate loci (rs7867029, rs7174015, rs12870438 and rs724078) for human male fertility traits, identified in a genome-wide association study (GWAS) of a Hutterite population in the USA, associated with male infertility in a Japanese population? rs7867029, rs7174015 and rs12870438 are significantly associated with the risk of male infertility in a Japanese population. Recently, a GWAS of a Hutterite population in the USA revealed that 41 single-nucleotide polymorphisms (SNPs) were significantly correlated with family size or birth rate. Of these, four SNPs (rs7867029, rs7174015, rs12870438 and rs724078) were found to be associated with semen parameters in ethnically diverse men from Chicago. This is a case-control association study in a total of 917 Japanese subjects, including 791 fertile men, 76 patients with azoospermia and 50 patients with oligozoospermia. Azoospermia was diagnosed on the basis of semen analysis (the absence of sperm in ejaculate), serum hormone levels and physical examinations. Oligozoospermia was defined as a sperm concentration of <20 × 10(6)/ml. We excluded patients with any known cause of infertility (i.e. obstructive azoospermia, varicocele, cryptorchidism, hypogonadotropic hypogonadism, karyotype abnormalities or complete deletion of AZF a, b or c). The SNPs rs7867029, rs7174015, rs12870438 and rs724078 were genotyped using DNA from peripheral blood samples and either restriction fragment length polymorphism PCR or TaqMan probes. Genetic associations between the four SNPs and male infertility were assessed using a logistic regression analysis under three different comparative models (additive, recessive or dominant). The genotypes of all four SNPs were in Hardy-Weinberg equilibrium in the fertile controls. The SNPs rs7867029 and rs7174015 are associated with oligozoospermia [rs7867029: odds ratio (OR) = 1.70, 95% confidence interval (CI) = 1.07-2.68, P = 0.024 (log-additive); rs7174015: OR = 6.52, 95% CI = 1.57-27.10, P = 0.0099 (dominant)] and rs12870438 is associated with azoospermia (OR = 10.90, 95% CI = 2.67-44.60, P = 0.00087 (recessive)] and oligozoospermia [OR = 8.54, 95% CI = 1.52-47.90, P = 0.015 (recessive)]. The association between rs7174015 and oligozoospermia under a dominant model and between rs12870438 and azoospermia under additive and recessive models remained after correction for multiple testing. There were no associations between rs724078 and azoospermia or oligozoospermia. Even though the sample size of case subjects was not very large, we found that three SNPs were associated with the risk of male infertility in a Japanese population. The three infertility-associated SNPs may be contributing to a quantitative reduction in spermatogenesis. This study was supported in part by the Ministry of Health and Welfare of Japan (1013201) (to T.I.), Grant-in-Aids for Scientific Research (C) (23510242) (to A.Ta.) from the Japan Society for the Promotion of Science, the European Union (BMH4-CT96-0314) (to T. I.) and the Takeda Science Foundation (to A.Ta.). None of the authors has any competing interests to declare.


Y chromosome haplogroup D2a1 is significantly associated with high levels of luteinizing hormone in Japanese men

The association between the Y chromosome haplogroup D2 and risk of azoospermia and low sperm motility has been previously studied, and it was indicated that haplogroups DE (YAP lineage) are associated with prostate cancer risk in Japanese males. Our assumption had been that Y chromosome haplogroups may be associated with sex hormone levels, because sex hormones have been deemed responsible for spermatogenesis and carcinogenesis. In this study, we assessed the association between Y chromosome haplogroups and sex hormone levels, including those of testosterone, sex hormone-binding globulin (SHBG), follicle-stimulating hormone (FSH), luteinizing hormone (LH), inhibin-B, and calculated free testosterone (cFT), in 901 young men from the general Japanese population (cohort 1) and 786 Japanese men of proven fertility (cohort 2). We found that the haplogroup D2a1 was significantly associated with high LH levels in a combined analysis involving two cohorts (β = 0.068, SE = 0.025, p = 0.0075), following correction for multiple testing. To date, this result is the first evidence that implicates Y chromosome haplogroups in an association with sex hormone levels.


Overview of genetic variation in the Y chromosome of modern Japanese males


The role of dipeptidyl peptidase 4 (DPP4) in the preservation of renal function: DPP4 involvement in hemoglobin expression

In a previous study, we demonstrated that dipeptidyl peptidase 4 (DPP4)-deficient rats were susceptible to reduced glomerular filtration rate as a result of streptozotocin (STZ)-induced diabetes. Therefore, we proposed that DPP4 might be responsible for the preservation of renal function. In this study, to verify the role of DPP4 in the preservation of renal function, we performed a microarray analysis of the kidneys of WT and DPP4-deficient rats after STZ treatment, and gene expression analysis using rat kidneys, human embryonic kidney 293 (HEK293) cells, and human renal cancer cells (CakI-1). The microarray analysis indicated that the expression levels of the transporter activity, heme-binding, and pheromone binding-related genes changed significantly. The results of gene expression analysis indicated that there were no significant differences in the expression levels of hemoglobin mRNA between the DPP4-deficient and WT rats; however, the expression levels of hemoglobin mRNA in the kidneys of DPP4-deficient rats tended to decrease when compared with those of both the non-STZ-treated and STZ-treated WT rats. The expression levels of hemoglobin in HEK293 and Caki-1 cells were significantly decreased when DPP4 was knocked down by siRNA, were significantly increased by the addition of soluble human DPP4, and were also significantly increased by the addition of the DPP4 inhibitor, sitagliptin. The expression level of DPP4 was also significantly increased by the addition of sitagliptin in both cell types. Our findings indicate that DPP4 regulates the expression of the hemoglobin genes, and might play a role in the preservation of renal function; however, the underlying mechanism of this preservation remains to be elucidated.


Development of a fluoride-responsive amide bond cleavage device that is potentially applicable to a traceable linker


Y chromosome gr/gr subdeletion is associated with lower semen quality in young men from the general Japanese population but not in fertile Japanese men

Several case-control studies have investigated whether Y chromosome haplogroups or deletions are associated with spermatogenic failure. However, the relationships between Y chromosome haplogroups or deletions and semen quality in general population have not been elucidated. In this study, we assessed relationships between Y chromosome haplogroups or deletions and semen parameters in 791 fertile Japanese men and 1221 young men from the general Japanese population. We found that the haplogroup D2 (M55 lineage) was significantly associated with lower semen parameters, especially total motile sperm count (P = 0.00051, beta = -0.097), in men from the general population but not in fertile men. In addition, we found that the gr/gr subdeletion was associated with semen quality and in particular, strongly associated with decreased sperm motility (P = 0.00041, beta = -3.14) and total motile sperm count (P = 0.00031, beta = -0.099) in men from the general population but not in fertile men. The combined analysis of fertile Japanese men and men from the general Japanese population showed that the haplogroup D2 (M55 lineage) and the gr/gr subdeletion were strongly associated with reduced sperm motility (P = 0.00056, beta = -2.71, and P = 7.7 × 10(-5), beta = -3.05, respectively) and that haplogroup O2b1 was strongly associated with elevated sperm motility (P = 0.00089, beta = 2.94). These observations add further support for the view that the gr/gr subdeletion diminishes sperm motility that consequently may result in male infertility.


Meta-Analysis of Interleukin Polymorphisms and NSAID Usage Indicates Correlations to the Risk of Developing Cancer

International Journal of Genomic Medicine


A meta-analysis of the association of PPARγ rs1801282 polymorphism and NSAID usage with the risk of developing cancer

Use of nonsteroidal anti-inflammatory drugs (NSAIDs) is correlated with a reduced risk of cancer through the reduction of inflammation, which is an important risk factor. Several studies have investigated polymorphisms in the peroxisome proliferator-activated receptor gamma (PPARγ) gene and NSAID use in association with cancer risk. However, these studies yielded mixed results. Therefore, we performed a meta-analysis to evaluate the association of PPARγ polymorphisms and NSAID usage with cancer risk. We conducted a comprehensive search of PubMed through May 2013. Odds ratios (ORs) with corresponding 95% confidence intervals (CIs) were calculated using the fixed-effect or random-effect model. A comprehensive search of the database revealed 6 studies that fulfilled the inclusion criteria. NSAID use was significantly associated with decreased cancer risk regardless of PPARγ rs1801282 genotypes. In a stratified analysis by cancer type, NSAID users who were minor allele carriers had significantly decreased colon cancer risk compared to non-NSAID users (OR=0.73, 95% CI=0.57-0.93), whereas NSAID users homozygous for the major allele had significantly decreased risk for cancers other than colon cancer compared to non-NSAID users (OR=0.79, 95% CI=0.69-0.91). Our results suggest that the association of PPARγ rs1801282 polymorphism and NSAID use with the risk of cancer may differ according to cancer type.


Development of a traceable linker containing a thiol-responsive amino acid for the enrichment and selective labelling of target proteins

A traceable linker that is potentially applicable to identification of a target protein of bioactive compounds was developed. It enabled not only thiol-induced cleavage of the linker for enrichment of the target protein but also selective labelling to pick out the target from contaminated non-target proteins for facile identification.


A Meta-Analysis of PTGS1 and PTGS2 Polymorphisms and NSAID Intake on the Risk of Developing Cancer

Several studies have investigated whether the polymorphisms in the prostaglandin endoperoxide synthase 1 (PTGS1) and PTGS2 genes and nonsteroidal anti-inflammatory drug (NSAID) use are associated with cancer risk; however, those studies have produced mixed results. Therefore, we performed a meta-analysis to evaluate the association between the PTGS1 and PTGS2 polymorphisms and the effect of NSAID use on the risk of developing cancer. We conducted a comprehensive search in PubMed through March 2012. The odds ratios (ORs) with the corresponding 95% confidence intervals (CIs) were calculated using the fixed-effect model or the random-effect model. The database search generated 13 studies that met the inclusion criteria. For PTGS1 rs3842787, NSAID users homozygous for the major allele (CC) had a significantly decreased cancer risk compared with non-NSAID users (OR = 0.73, 95% CI = 0.59-0.89). For PTGS2 rs5275 and rs20417, there were no significant differences between the gene polymorphism and NSAID use on cancer risk among the 8 and 7 studies, respectively. However, in the stratified analysis by the type of cancer or ethnicity population, NSAID users homozygous for the major allele (TT) in rs5275 demonstrated significantly decreased cancer risk compared with non-NSAID users in cancer type not involving colorectal adenoma (OR = 0.70, 95% CI = 0.59-0.83) and among the USA population (OR = 0.67, 95% CI = 0.56-0.82). NSAID users homozygous for the major allele (GG) in rs20417 displayed a significantly decreased cancer risk than non-NSAID users among the US population (OR = 0.72, 95% CI = 0.58-0.88). For the PTGS2 rs689466 and rs2745557 SNPs, there were no significant differences. This meta-analysis suggests that the associations between PTGS polymorphisms and NSAID use on cancer risk may differ with regard to the type of cancer and nationality.


Y chromosome haplogroup D2* lineage is associated with azoospermia in Japanese males

Several studies have investigated whether particular Y chromosome haplogroups are associated with spermatogenic failure in Japanese males; however, they produced differing results. In this study, to investigate the association of Y chromosome haplogroup with spermatogenic failure, we recruited 451 infertile patients and 730 fertile men from a Japanese population and typed their Y chromosome haplogroups. The infertile patients were suffering from varicocele, azoospermia, oligozoospermia, asthenozoospermia, obstructive azoospermia, karyotype abnormalities, microdeletions of the long arm of the Y chromosome, or other conditions that affect fertility. The frequency of haplogroup D2* was significantly higher (odds ratio = 2.28, 95% confidence interval = 1.44-3.61, P = 0.00034 using chi-square test) among the men with azoospermia than among the fertile men. None of the other Y haplogroups displayed associations with particular types of infertility. In conclusion, Y chromosome haplogroup D2* is associated with spermatogenic failure in Japanese males, suggesting that the Y chromosome lineage can have significant effects on spermatogenesis.


Replication study and meta-analysis of human nonobstructive azoospermia in Japanese populations

Recently, a Chinese genomewide association study (GWAS) identified four autosomal single-nucleotide polymorphism (SNP) loci as being significantly associated with risk factors for nonobstructive azoospermia (NOA; P < 5 × 10(-8)). In the present study, we performed a replication study on two Japanese cohorts from different institutions in order to evaluate whether SNP loci are associated with NOA. The four SNPs (rs12097821, rs2477686, rs10842262, and rs6080550) reported in the Chinese GWAS were genotyped in 490 NOA patients and 1167 controls. To assess the significance of the associations between each of the four SNPs and NOA in the Japanese population, the association results for the two cohorts were combined by meta-analysis. In the meta-analysis, the combined per-allele odds ratios (ORs) for the four SNPs and their respective 95% confidence intervals (CIs) were as follows: rs12097821, OR = 1.10 (CI = 0.89-1.37); rs2477686, OR = 1.11 (CI = 0.87-1.43); rs10842262, OR = 1.11 (CI = 0.94-1.32); and rs6080550, OR = 0.96 (CI = 0.76-1.21). None of the SNPs was significantly associated with NOA (P > 0.05). However, three of four SNPs (rs12097821, rs2477686, and rs10842262) showed associations in the same direction in Japanese men as those reported in the Chinese GWAS. To determine whether the four SNPs are genetic risk factors for NOA, the effect sizes of NOA risk factors require further investigation using larger independent sets of case-control samples of populations, including Japanese and Chinese populations.


地域薬局薬剤師による糖尿病自己管理支援におけるポートフォリオ活用の試み

地域薬局薬剤師による糖尿病自己管理支援におけるポートフォリオ活用の試み

日本ファーマシューティカルコミュニケーション学会会誌

日本ファーマシューティカルコミュニケーション学会会誌


A Questionnaire Survey on Medication Adherence at a Community Pharmacy in Okayama to Determine the Requirements for Pharmaceutical Care

Japanese Journal of Pharmaceutical Health Care and Sciences


Intrathecal Disposition of ARTCEREB® Irrigation and Perfusion Solution for Cerebrospinal Surgery in Rats

We investigated the disposition of ARTCEREB® irrigation and perfusion solution (Artcereb) during intrathecal perfusion in a lateral ventricle-cisternal perfusion model in conscious rats. In this perfusion model, the perfusion rate was set at 0.35 ml/kg/h, taking into consideration the clinical perfusion rate (500 ml/60 kg/d). The influence of Artcereb on electrolytes in cerebrospinal fluid (CSF) and blood were then investigated. After 24 h of ventriculocisternal perfusion with Artcereb using the push-pull method, output of K(+), Na(+) and Cl(-) to the cistern magna was very similar to input of these electrolytes in Artcereb infused intraventricularly. Recovery rates of K(+), Na(+) and Cl(-) after perfusion were 102%, 105% and 100% when calculated using the recovered perfusion solution. In addition, concentrations of K(+), Na(+) and Cl(-) in blood remained almost constant at near baseline levels throughout perfusion. Thus, intrathecally perfused Artcereb did not affect electrolyte balance in the CSF and blood. To confirm the dynamics of Artcereb distribution, a whole body autoradiography study was performed at 1 and 6 h after perfusion with ¹ C-inulin-added Artcereb. Radioactivity was detected in the entire CSF space of the brain, and the cribriform plate in the nasal cavity, and the cerebrospinal cavity. Radioactivity was observed in the bladder, thus suggesting that some ¹ C-inulin was transferred to the bloodstream via a physiological route, and was excreted renally.


Pharmacological assessment of ARTCEREB® irrigation and perfusion solution for cerebrospinal surgery using primary cultures of rat brain cells

ARTCEREB irrigation and perfusion solution (Artcereb), an ethical pharmaceutical, is typically applied inside the skull and spinal cavity as artificial fluid. Artcereb is composed of glucose and electrolytes (Na+, K+, Mg2+, Ca2+, Cl-, HCO3- and P) and has a pH of 7.3. An in vitro assessment of the effects of Artcereb on cell culture of rat fetal astrocytes or rat fetal brain cells was performed in comparison with normal saline and lactated Ringer's solutions. Furthermore, the effects of Artcereb on cell culture of rat fetal brain cells were also assessed in comparison with Krebs bicarbonate solution. Cell function after exposure to Artcereb was assessed based on 3H-thymidine incorporation activity. Cell function after exposure to Artcereb and lactated Ringer's solution in primary cultures of rat fetal astrocytes remained unaffected when compared to that after exposure to normal saline. Cell function after exposure to Artcereb in a primary culture of rat brain cells remained unaffected as compared to that after exposure to normal saline and lactated Ringer's solution. However, function decreased after exposure to a modified Artcereb formulation lacking bicarbonate, thus confirming that the presence of bicarbonate is essential for the Artcereb formulation.


Consultation clinics for complementary and alternative medicine at Japanese university hospitals: An analysis at Tokushima University Hospital

Here, we report on a Consultation Clinic for Complementary and Alternative Medicine (CAM) which we established at Tokushima University Hospital in July of 2007 with the aim of providing person-to-person information on CAM, though not CAM therapy itself. In December of 2008, we received 55 applications for consultation, 37% concerning health foods, 37% Japanese herbal medicine (Kampo), and 26% various other topics. The consultants (nutritionists and pharmacists) communicated individually with 38 applicants; malignancies (26%) and cardiovascular disease (24%) were the main underlying concerns. To promote the quality of consultation, data was collected by means of focus group interviews concerning the perspective of the consultants. Safe and effective use of CAM requires a network of communication linking individuals, consultation teams, physicians, primary care institutions and university hospitals. To advance this goal, we plan to broaden the efforts described herein. Our findings indicate that the specific role of the consultation clinic in promoting the scientific use of CAM merits further study.


Perfusion fluids used in neurosurgery affect cerebrospinal fluid and surrounding brain parenchyma in the rat ventriculocisternal perfusion model

ARTCEREB, an irrigation and perfusion solution (Artcereb), is a preparation intended for the irrigation and perfusion of the cerebral ventricles, and it is therefore important to evaluate its effects on cerebrospinal fluid (CSF) and on the surrounding cerebrospinal parenchyma. To confirm the kinetics of the perfusion fluid component, we performed whole body autoradiography and examined glucose balance during ventriculocisternal perfusion with (14)C-glucose labeled Artcereb in the rat model, which simulates ventricular irrigation or ventriculocisternal perfusion in clinical neurosurgery. We also performed ventriculocisternal perfusion with Artcereb, lactated Ringer's solution, or normal saline, and observed the effect of these solutions on animal condition and on brain tissue morphology. In the kinetic study, diffusion of (14)C-glucose from the perfused Artcereb to the cerebrospinal tissue was seen on whole body autoradiography, and almost 90% of the glucose in the perfusion fluid was distributed to the cerebrospinal tissue and the systemic circulation. These data indicated that the perfusion fluid interacted actively with the CSF, surrounding parenchyma and the systemic circulation, and suggested that the formation of perfusion fluid affected CSF composition and cerebrospinal tissue functions. Animals perfused with normal saline were associated with serious symptoms including tonic convulsions and death, and exhibited neuronal death in the cerebrum. However, these severe changes were not observed in animals perfused with Artcereb or lactated Ringer's solution. We therefore propose that during neurosurgery, it is extremely important to use a physiological solution like Artcereb which closely resembles normal human CSF, in order to maintain cerebrospinal function and to alleviate postoperative adverse events.


ARTCEREB® Irrigation and Perfusion Solution for Cerebrospinal Surgery:Pharmacological Assessment Using Human Astrocytes Exposed to Test Solutions

アートセレブ®脳脊髄手術用洗浄灌流液:ヒトアストロサイト培養系を用いた薬理学的評価

ARTCEREB® irrigation and perfusion solution (Artcereb) is a preparation intended for the irrigation and perfusion of the cerebral ventricles, and it is therefore important to evaluate the effects of Artcereb on brain cells. In vitro assessment of the effects of Artcereb in cell cultures of human fetal astrocytes was conducted in comparison with normal saline and lactated Ringers solution. The effects of exposure to Artcereb were evaluated based on microscopic images of the mitochondria stained with rhodamine 123. The effects of exposure to Artcereb on cell function were also evaluated by quantitative analysis of mitochondrial activity based on rhodamine 123 and 3H-thymidine incorporation. Morphological changes in nuclear structure were also evaluated. The results of the present study showed that cell function in cell cultures of human astrocytes was relatively unaffected by exposure to Artcereb as compared with normal saline or lactated Ringers solution, suggesting that Artcereb has less effect on brain cells than normal saline or lactated Ringers solution when used for the irrigation or perfusion of the cerebral ventricles.

Journal of the Pharmaceutical Society of Japan


Synthesis and Pharmacological Evaluation of New 1-Hydroxy-1-phenyl-2, 3,4, 5-tetrahydro-1H-3-benzazepines as NE Potentiators

4-Hydroxy-2-methyl-4-phenyl-1,2,3,4-tetrahydroisoquinoline (PI-OH) (1a) and its derivatives form a new class of compounds which possess norepinephrine (NE) potentiating activity. As a new series of compounds, 1-hydroxy-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepines (2a--f) were synthesized by the intramolecular Barbier reaction of N-[2-(2-iodophenyl)ethyl]phenacylamines (6a--f) with n-BuLi as a key reaction step. The potentiating activities of the benzazepines 2a--f on the contraction of rat anococcygeus muscle induced by NE were tested. Among the compounds tested in this study, compound 2a showed moderate potentiating activity (the activity ratio was 7.3-fold at 3 x 10(-5) M).


Quantitative Structure-Activity Relationship for Inhibition of CYP2B6 and CYP3A4 by Azole Compounds - Comparison with Their Binding Affinity


医薬品安全性情報コミュニティ·データベースシステム:妊娠と薬に関する安全性情報の共有に向けて

Journal of Information Processing and Management


Construction of a model for water purification mechanisms in a river by using a neural network approach

A model for purification mechanisms in a river was proposed to express changes in the values of BOD (Biochemical Oxygen Demand), COD (Chemical Oxygen Demand), T-N (Total Nitrogen), and T-P (Total Phosphorous) as the combination of inflows, streams, and weirs. Because expressions for these functions are unknown, the model first constructs multi-layer neural-network functions based on observations, and then uses the derivatives to evaluate the cause and effect of pollution.For data of the Tamagawa through Tokyo, Japan in 2002, the model suggested that the cause of pollution is inflows from sewage, the stream of the Tamagawa has purification functions for COD and T-P, but has little ability for T-N, and the weirs have purification for COD, but have no purification for T-N and T-P. The results from the model were consistent with common sense for the water quality, and thus, there was no failure in the model.


Prescribing patterns during pregnancy at Tokushima University Hospital

The purpose of this study was to investigate the drugs prescribed for pregnant women at Tokushima University Hospital in Japan, and to evaluate the risks associated with the use of these drugs during pregnancy according to the pregnancy risk classification systems of the PDA, ADEC and that used in manufacturers' package inserts. Of 496 female outpatients who gave birth at Tokushima University Hospital, 81% had been prescribed at least one drug, and the mean was 4.0 drugs per patient. Analysis of the prescriptions revealed a pattern of drug use very different from that in other countries, in that Kampo (Japanese herbal) medicines were also prescribed. For more than half of the oral drugs prescribed, risks were not given in the PDA and/or ADEC pregnancy risk systems. In addition, there was no listing of pregnancy risks in the package inserts of 20 drugs. We therefore consider it to be very important to develop a globally harmonized system for evaluating the risks associated with prescription drugs taken during pregnancy.

Japanese Journal of Pharmaceutical Health Care and Sciences


A structural similarity evaluation by SimScore in a teratogenicity information sharing system

Many efforts have been devoted to the development of computer-aided prediction of drug toxicity over the decades, but at present, systems and programs available for predicting teratogenicity from chemical structures do not always give satisfactory answers yet, mainly because of the complex and unknown mechanism of reproductive and developmental toxicity. We developed a novel algorithm and implemented in the program "SimScore" to evaluate quantitatively the structural similarity score of a target compound with the teratogenic drugs which are defined as serious human teratogens by the United States Food and Drug Administration. In SimScore, a molecular structure is divided into its skeletal and substituent parts in order to perform similarity comparison for these parts independently. This idea is based on that compounds with the same or similar skeleton show a similar biological activity, but their activity strengths depend on the variation of substituents. We demonstrated the usefulness of SimScore by applying it to an example.SimScore will be used in our web-based information system about teratogenicity to predict the potential risk of query compounds.


ニューラルネットワークを用いたドナウ川の水質解析-OECDレポートを用いて-

ニューラルネットワークを用いたドナウ川の水質解析-OECDレポートを用いて-

We researched the water quality of the Donau based on OECD reports. Observations in the reports have defects that make evaluation of environmental situations difficult. By using interpolations in the compensation quantitative structure-activity relationships (CQSAR), we complement the defects in the water quality of the rivers. Thus, we get a complete data set for dissolved oxygen, biochemical oxygen demand, and total phosphorus. Using the data set, we examine re-naturalization of the Donau. We investigate the effect of dams between Slovakia and Hungary, by using reconstructions of neural networks in CQSAR. The water quality of the Donau has been improved as to DO and BOD. The rivers have been re-naturalized. We believe that the effect is found in these indexes; however, not so much for the T-P. We find a negative effect of the dam construction on changes of DO in the Hungary Donau.

OECDによるドナウ川の水質データの経年変化に関し，欠測データを含むデータの解析が可能な多階層型ニューラルネットワークシミュレーション(CQSAR)法を用いて，欧州の河川再自然化政策等による河川の水質浄化の有効性およびダム建設の影響を検討した．ドイツ圏で施行されている河川再自然化や水質浄化政策はT-P値の改善にはあまり効果がなかったが，DO,BOD値を改善していた．特に，ドナウ川のハンガリー流域でのBOD値に浄化傾向がみられた．ドナウ川のBOD値から自然浄化作用が河川浄化策の施行により回復してきていると判断できた． スロヴァキアとハンガリー間のドナウ川に設けられたダム(ガブチコボ・ナジュマロシュ・ダム)の影響は，DO値は悪化される傾向にあり，逆にBOD値は改善される傾向にあった．また，ダム建設によりダム下流のドナウ川のハンガリー流域でのDO値やBOD値が，ダム上流域の水質と関係しなくなったことが明らかになった．


Extraction of a parameter as an index to assess water quality of the Tamagawa, Tokyo, Japan, by using neural networks and multivariate analysis

Parameters as an index to efficiently assess the pollution level of the upper, middle, and lower streams of the Tamagawa (Tokyo, Japan) based on measured water quality were determined by using multivariate analysis, principal component analysis (PCA), and cluster analysis (CA) for data measured from 1994 to 2002. Missing data during 2000-2002 were estimated using a perceptron type neural network and arithmetic progression. The combination of scores for the first and second principal components obtained by PCA enabled classification of the upper, middle, and lower streams of the Tamagawa. The CA results corresponded well with the PCA results. Based on the score of the first principal compornent determined here, contributions to the water pollution of the middle and lower streams should be decreased to improve the water quality of the Tamagawa.


線形多変量解析を用いた徳島吉野川の水質の解析

線形多変量解析を用いた徳島吉野川の水質の解析

Principal component analysis, cluster analysis and neural network were applied to assess the pollution levels along the Yoshinogawa River in Tokushima, Japan, by using water quality data (Dissolved Oxygen (DO), Biochemical Oxygen Demand (BOD), Chemical Oxygen Demand (COD),and Total Phosphorus (TP)) measured in 2002. Because there are a few defects in the data, we estimated them by linear equation and neural network to avoid information loss.In the Yoshinogawa River, there are four branching bays (Figure 1). Though it is usually difficult to find the relationships between the water quality of the Yoshinogawa River and the distances from their estuaries, the first principal components consisting of five parameters explained the nature of the river well. The water quality at Heiwabashi of the Shinmachigawa River was the most contaminated in the Yoshinogawa River (Figure 6, Table 12). This suggests that there is a source of water pollution in the upper region of Heiwabashi and that there is a source to improve water quality in the lower region of Heiwabashi.

吉野川の水質浄化の指針を得るために，吉野川水系の上流から河口に至るまでの5種類の水質パラメータの変化を主成分分析とクラスター分析を用いて解析し，吉野川水系の上流から下流までの汚染度をみるための複合的な指標を得ることを試みた．吉野川の汚染の傾向を見るためには，少なくとも5つのパラメータほぼ同じ程度重要であった．吉野川は4つの河口を持つ(Figure 1)が，水質は河口からの距離とは対応しておらず(Table 5)，最も河口からの距離が長い平和橋(銅山川)と河口の1つである新町川の水質汚染が進んでいることがわかった(Figure 6，Table 12)．このことから，それぞれの流域の水質に影響を与えている要因を個々に探る必要があることが示唆された．吉野川の水質浄化のためには，このような要因の解析と総合的な環境指標に基づいた解析が重要であることがわかった．


Quantitative structure - activity relationship study of binding affinity of azole compounds with CYP2B and CYP3A


「妊娠と薬」に関する意識調査-薬剤師, 薬学生, 研究者の場合

「妊娠と薬」に関する意識調査-薬剤師, 薬学生, 研究者の場合

The aim of this study was to assess the awareness of pharmacists, pharmacy students and researchers regarding "Drugs and Pregnancy". In order to do this, a questionnaire was given to 211 attendees of a symposium held under a pharmacists' education program held at the University of Tokushima. The response rate was 108/211 (51%) and the breakdown of the respondents was students 46%, pharmacists 36% and researchers 9%. Among the 36 persons with experience of delivery, 67% had taken medicine during pregnancy, and 42% of them were worried about the embryotoxic effects of drugs. Persons with no experience of delivery were more worried about such effects than those who had experienced delivery. Before being given a prescription, 71% of the respondents had been asked about the possibility of pregnancy by a physician. On the other hand, no respondent had been asked about this by a pharmacist, suggesting that pharmacists are passive with regard to providing information about drugs and pregnancy. Information on drugs and pregnancy was mainly obtained from drug package inserts, books and pharmaceutical companies. The majority of the respondents (96%) were interested in a web site from which they could retrieve information on drug safety during pregnancy. They showed a willingness to cooperate with regard to recording medicines used during the pregnancy as well as the outcome of their pregnancies. Eighty-nine percent of the researchers and 78% of the students wanted to use teratogenicity information in drug design. All of the researchers answered that it was beneficial to share information on reproductive and developmental toxicity. The results of our questionnaire survey showed that many people desired drug safety information during pregnancy, and were willing to share such information.


Effect of advanced pharmaceutical services for chemotherapy on length of hospital stay

We prepared a chemotherapy information leaflet that helped to enhance patients' understanding of chemotherapy and to reduce their anxiety about its adverse reactions. The aim of the present study was to evaluate pharmaceutical services with regard to the management of adverse reactions, length of hospital stay and the medical costs for women with gynecologic cancer who received chemotherapy. We carried out a retrospective chart survey of 28 ovarian cancer patients to clarify the effects of pharmaceutical care on their chemotherapy. Half of the patients were given chemotherapy information verbally and were assigned to Group A, and the other half were supplied with an information leaflet and a self-check list for adverse reactions and assigned to Group B. There were no significant differences between the two groups regarding the occurrence of adverse events, except for fatigue and insomnia. However, 50% (7/14) of the patients in Group A stayed longer in hospital to be free from adverse reactions, especially myelosuppression. The mean hospital days per cycle of chemotherapy in Group B (2.76±0.80 days) was significantly shorter than that in Group A (7.28±3.97 days, p<0.001). Thus, our pharmaceutical care using chemotherapy information leaflets had a favorable effect in terms of shortening the length of hospital stay.


Prescription drug use during pregnancy in Japan


Soybean oil fat emulsion to prevent TPN-induced liver damage:Possible molecular mechanism and clinical implications

Long-term total parenteral nutrition (TPN) is known to be associated with cholestasis and hepatic steatosis, which can be lethal in infants who cannot be fed orally. The present review focuses on the metabolic complications in the liver that may occur due to the excessive administration of fat-free TPN. We have recently developed an infant rat model of hepatic dysfunction and steatosis induced by overdose of fat-free TPN. In this model, plasma levels of liver enzymes in the fat-free TPN group were found to be significantly higher than in the other groups (i.e., the oral diet and fat-containing TPN groups). Pathological examination showed hepatomegaly and severe fatty changes without cholestasis in the liver of infant rats that received fat-free TPN. We clearly demonstrated that the addition of soybean oil emulsion to the TPN regimen prevented hepatic dysfunction and fatty changes. In the present review, we discuss the molecular mechanism of the hepatic dysfunction induced by fat-free TPN and the role of soybean oil fat emulsion in the TPN regimen. We also discuss the clinical implications of soybean oil-containing TPN solutions and point out the importance of including fat in the TPN regimen in order to prevent the hepatic abnormalities associated with the excessive administration of fat-free TPN.


Pharmacotherapy During Pregnancy-Influence on both the Mother and the Fetus-

妊娠期の薬物療法∼その実態と母児への影響∼


A new synthesis of phenolic 1-hydroxy-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepines


Determination of lipophilicity by reversed-phase high-performance liquid chromatography: Influence of 1-octanol in the mobile phase

Lipophilicity was evaluated using a novel RP-HPLC stationary phase (Discovery-RP-Amide-C16) with and without 1-octanol added to the mobile phase. A set of 46 drugs and flavonoids characterized by a broad structural diversity and a wide log Poct range (-0.69 to 5.70) was selected for this study. This set consists of neutral solutes and solutes with acidic or ampholytic functionalities which were maintained neutral at pH 2.5 or 4. In our conditions, the addition of 1-octanol in the mobile phase proved a key factor to derive a lipophilicity index log k(w) highly correlated with log Poct for all investigated solutes. 1-Octanol improved the correlation between log Poct and log k(w) mainly by influencing the retention behavior of the solutes with log Poct values below +3. This study brings additional evidence that under proper experimental conditions of stationary and mobile phases, RP-HPLC is a very useful method to obtain log Poct values.


Clinical QSAR study on the prediction of drug excretion into human breast milk


Major role of soybean oil in preventing decrease in CYP4A1 mRNA content relative to PPARαmRNA down-regulation in infant rats

Changes in the levels of CYP4A1, PPARalpha, and RXRalpha mRNA expression in the liver following overdose of fat-free or fat-containing total parenteral nutrition (TPN) were studied in 3-week-old male Sprague-Dawley rats. The rats were divided into three groups: group 1, oral diet; group 2, fat-free TPN; and group 3, TPN with 20% of calories from soybean oil emulsion. Levels of CYP4A1, PPARalpha, and RXRalpha mRNA in the fat-free TPN group were significantly lower than those in the other groups. Levels of CYP4A1 and PPARalpha mRNA were strongly correlated (r=0.849), and levels of CYP4A1 and RXRalpha mRNA were weakly correlated (r=0.618). This is the first report of a strong correlation between the levels of CYP4A1 and PPARalpha mRNA following overdose of fat-free or fat-containing TPN in infant rats. Our results also indicate that it is important to include fat in TPN regimens in order to prevent CYP4A1 mRNA down-regulation, which may be related to changes in PPARalpha mRNA levels in the liver.


Soybean oil in total parenteral nutrition maintains albumin and antioxidant enzyme mRNA levels

Changes in albumin and antioxidant enzyme mRNA expression in infant rat liver following administration of total parenteral nutrition (TPN) with/without soybean oil emulsion were studied. Infant rats were divided into three groups: group 1=oral diet, group 2=TPN without fat, and group 3=TPN with 20% of calories from soybean oil emulsion. The period of TPN administration was 4 d. Serum aspartate aminotransferase and alanine aminotransferase levels were higher in group 2 than in the other groups, with similar levels seen in the other groups. Albumin, Cu, Zn-superoxide dismutase, and glutaredoxin 1 mRNA expression levels were lower in group 2 than in the other groups, with similar levels seen in the other groups. Catalase mRNA expression was higher in group 1 than in the other groups, with the lowest level seen in group 2. Soybean oil emulsion should be included in TPN regimens to prevent down-regulation of albumin and antioxidant enzyme mRNA expression.


Defects in apoptotic signal transduction in cisplatin-resistant non-small cell lung cancer cells

Non-small cell lung cancer (NSCLC) often shows intrinsic multidrug resistance, which is one of the most serious problems in cisplatin-based adjuvant chemotherapy. Anticancer drugs exert at least part of their cytotoxic effect by triggering apoptosis. In order to understand the molecular alterations leading to heterogeneous cisplatin sensitivity and apoptosis inducibility in NSCLC cells, we analyzed various apoptotic pathways, including the activation of caspase-8, -9 and -3, the release of cytochrome c from mitochondria and the expression levels of pro- and anti-apoptotic proteins such as Bax, Bad, Bcl-2, Bcl-xL, Fas and p53 using heterogeneously apoptosis-sensitive cells (Ma-10, Ma-31 and Ma-46). Cisplatin treatment induced the activation of caspase-8, -9 and -3 and the release of cytochrome c in apoptosis-sensitive Ma-46. The expression of Bcl-xL was the highest and p53 was not expressed in apoptosis-resistant Ma-31, and Fas was not expressed in Ma-46. These expression levels were not correlated with the apoptosis inducibility of the three cell lines. These results suggest that blockage of the apoptotic signal from mitochondria is responsible for apoptosis resistance in NSCLC cell lines. Our findings also indicate that anti-apoptotic Bcl-xL and pro-apoptotic p53 are necessary but not sufficient for resistance to cisplatin-induced apoptosis in NSCLC cells.


徳島大学病院におけるTPN処方設計支援ソフトの使用評価と改良

徳島大学病院におけるTPN処方設計支援ソフトの使用評価と改良

Fluid therapy is a fundamental treatment for almost all inpatients, and nutritional therapy is the most important type of such therapy. Currently, total parenteral nutrition (TPN) is widely used to provide complete nutritional support but 30-40% of TPN recipients are considered to in a malnourished state due to inappropriate TPN prescriptions. Computer-assisted TPN prescription systems are thought to be useful to clinical professionals in solving this problem because they can perform complicated calculations that make it easy to draw up correct TPN prescriptions. So, we made a prototype system for aiding the preparation of TPN prescriptions based on Microsoft^[ !R] EXCEL and it was tested by physicians, pharmacists, dietitians and nurses engaged in TPN therapy at Tokushima University Hospital. We also had them fill out a questionnaire on the system in which they rated it for ease of operation and usefulness according to a scoring system in which 5 indicated full marks. Our Computer-Assisted TPN Prescription System was improved on the basis of the questionnaire results and is now being effectively used by the nutrition support team (NST) in Tokushima University Hospital.

Japanese Journal of Pharmaceutical Health Care and Sciences


Expression of multidrug resistance proteins and accumulation of cisplatin in human non-small cell lung cancer cells

In order to understand and overcome multidrug resistance (MDR) of human non-small cell lung cancer (NSCLC), mRNA and protein expression levels of P-glycoprotein (MDR1), multidrug resistance-associated protein 1 (MRP1), and lung resistance-related protein (LRP) were investigated and compared with the chemosensitivity and the intracellular/intranuclear cisplatin accumulation of three NSCLC cell lines (Ma-10, Ma-31, and Ma-46). Ma-31 was more resistant than Ma-10 and Ma-46 to cisplatin, carboplatin, etoposide, and paclitaxel. The mRNA level of MDR1 was extremely low, and MDR1 protein was not detected in all cell lines. MRP1 mRNA expression was highest in Ma-31 and lowest in Ma-10, but there was no notable difference between the MRP1 protein expression in three cell lines. LRP mRNA/protein was equally expressed in Ma-10 and Ma-31, but was nominal in Ma-46. The intracellular/intranuclear cisplatin accumulation of the cells was determined to be Ma-31>Ma-46>Ma-10. Thus, MDR1, MRP1, and LRP mRNA and protein expression levels were not correlated with the chemosensitivity or the intracellular/intranuclear cisplatin accumulation of each cell line. The present results indicate that MDR proteins (MDR1, MRP1, and LRP) may not play an important role in the chemoresistance and drug efflux of NSCLC cells.


Role of soybean oil fat emulsion in the prevention of hepatic xenobiotic transporter mRNA up-and- down -regulation induced by overdose of fat-free total parenteral nutrition in infant rats

Total parenteral nutrition (TPN) is associated with cholestasis and hepatic steatosis in human infants. The present study focused on the changes in hepatic xenobiotic transporters associated with overdose of fat-free or fat-containing TPN in infant rats. Three-week-old male Sprague-Dawley rats were divided into three groups: group 1 received an oral diet, group 2 received TPN without fat, and group 3 received TPN with 20% of its calories from fat (soybean oil emulsion). After TPN administration for 4 days, both serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels, which are indicators of hepatic dysfunction, in group 2 were significantly higher (p<0.001) than those in the other groups, whereas there were no differences between groups 1 and 3 in either serum AST or ALT levels. The serum bilirubin concentration in group 2 was also markedly higher than that in the other groups. Mdr2, Bsep, Mrp2, Mrp6, Oct1, and Oat2 mRNA levels were decreased in group 2 (fat-free TPN) compared with those in group 1 (oral diet), whereas Mdr1b, Mrp1, and Mrp5 mRNA levels were increased. Specifically, the level of Mdr1b mRNA in group 2 was 16 times higher (p<0.001) than that in group 1. On the other hand, the changes in these mRNA expression levels in group 3 (fat-containing TPN) were smaller than those in group 2, and specifically, the expression levels of Mdr1b, Mrp1, Mrp5, Mrp6, and Oat2 mRNA in group 3 were not significantly different from those in group 1. The results of the present study indicate that including fat in the TPN regimen is very important in preventing the mRNA up- and down-regulation of xenobiotic transporters, which is considered to be the main factor responsible for the abnormal hepatic changes such as cholestasis associated with the excessive administration of fat-free TPN.


Overexpression of constitutive signal transducer and activator of transcription 3 mRNA in cisplatin-resistant human non-small cell lung cancer cells.

Non-small cell lung cancer (NSCLC) often shows intrinsic multidrug resistance, which is one of the most serious problems in cisplatin-based adjuvant chemotherapy. Recently, the constitutive activation of signal transducer and activator of transcription (STAT) factors has been found in a variety of human cancers. In the present study, the mRNA expression of STATs in various human NSCLC cell lines was investigated by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) to determine whether STATs can be implicated in cisplatin resistance and apoptosis inducibility. Cisplatin triggered apoptosis in Ma-46 based on biochemical and morphological findings, but not in Ma-31. The mRNA expression of STAT3 was highest in cisplatin-resistant Ma-31 and lowest in cisplatin-sensitive Ma-46. A 6-hour exposure of cancer cells to cisplatin failed to stimulate STAT3 mRNA expression. Therefore, an increased transcriptional level of constitutive STAT3 may be related to the suppressive regulation of the apoptotic pathway in intrinsically chemo-resistant NSCLC cells.


Soybean oil fat emulsion prevents cytochrome P450 mRNA down-regulation induced by fat-free overdose total parenteral nutrition in infant rats.

Changes in the mRNA expression of hepatic cytochrome P450 (CYP) isoenzymes associated with overdose of fat-free or fat-containing total parenteral nutrition (TPN) were investigated in infant rats. Three-week-old male Sprague-Dawley rats were divided into three groups: group 1 received an oral diet, group 2 received TPN without fat, and group 3 received TPN with 20% of calories from fat (soybean oil emulsion). After TPN administration for 4 d, serum aspartate aminotransferase (AST) levels in group 2 were significantly increased (p<0.01) compared with the other groups. The mRNA expression of hepatic CYP isoenzymes in group 2 decreased to 0.76 to 31% of that in group 1 (p<0.01), but that in group 3 was maintained at 32 to 84% of that in group 1. These results indicate the importance of including fat in TPN regimens to prevent not only hepatic dysfunction but also mRNA down-regulation of liver CYP isoenzymes.


Novel antioxidants isolated from plants of the genera Ferula, Inula, Prangos and Rheum collected in Uzbekistan

We examined the effects of 48 compounds isolated from Ferula pallida, F. penninervis, Inula macrophylla, Prangos pabularia, P. tschimganica and Rheum maximowiczii collected in Uzbekistan on ADP/Fe2+-induced lipid peroxidation of egg yolk phosphatidylcholine liposomes. Of those compounds, 23 inhibited ADP/Fe2+-induced lipid peroxidation and nine showed especially strong inhibition of lipid peroxidation. Most compounds that inhibited peroxidation scavenged the 1,1'-diphenyl-2-picrylhydrazyl (DPPH) radical, indicating that the inhibition was due to radical scavenging. However, some compounds did not scavenge DPPH but inhibited lipid peroxidation significantly, suggesting that their inhibitory effect was not due to radical scavenging but to some other mechanism, such as prevention of Fe2+ function. Thus, we found various new antioxidants, some of which had a unique mechanism of action, in Ferula, Inula, Prangos and Rheum plants collected in Uzbekistan as seeds used in medicine.


Antioxidant effects of stereoisomers of N-acetylcysteine (NAC), L-NAC and D-NAC, on angiotensin II-stimulated MAP kinase activation and vascular smooth muscle cell proliferation

We examined the effects of the stereoisomers of N-acetylcysteine (NAC), L-NAC and D-NAC, on cellular glutathione (GSH) concentration and whether NAC-regulated cellular GSH levels are directly associated with angiotensin II (Ang II)-induced intracellular signaling events in vascular smooth muscle cells (VSMC). Both L-NAC and D-NAC similarly increased intracellular GSH concentration. We found that L-NAC and D-NAC both inhibited Ang II-induced c-Jun N-terminal kinase and p38 mitogen-activated protein kinase activation and [(3)H]-thymidine incorporation in VSMC. Our present study indicates the comparable effects of NAC stereoisomers in regulating intracellular GSH and the redox-dependent intracellular signaling mechanisms in VSMC.


医薬品安全性情報コミュニティの構築にむけて

医薬品安全性情報コミュニティの構築にむけて


Involvement of histone phosphorylation in apoptosis of human astrocytes after exposure to saline solution

We have previously found using inhibitors of protein phosphatase that phosphorylation of histones may be involved in thymocyte apoptosis. In this study, we examined whether histone modification occurs in astrocyte apoptosis induced by a pathological condition in the absence of drug. Incubation of cultured human astrocytes with growth medium for 24 h after exposure to saline solution for 30 min induced an increase in terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-positive cells and nuclear condensation, biochemical and morphological hallmarks of apoptotic cell death. Acetic acid-urea-Triton X-100 (AUT) gel electrophoresis of the nuclear histone fraction and N-terminal peptide analysis showed that the treatment with saline solution caused rapid changes in phosphorylation of H2A subfamilies, but not in histone acetylation. The phosphorylation of the two subtypes increased markedly, whereas the phosphorylation of one subtype decreased. In contrast, exposure to ACF-95, an artificial cerebrospinal fluid (CSF), was associated with little induction of apoptotic cell death and induced less changes in histone phosphorylation. These results support the previous idea that chemical modification of histones is involved in the DNA fragmentation in astrocytes undergoing apoptosis.


Construction of a drug safety information community in Japan


Lipophilicity measurement by reversed-phase high-performance liquid chromatography (RP-HPLC): A comparison of two stationary phases based on retention mechanisms


Construction of a Growing Information-Community for Teratogenic Agents

双方向·成長型薬物催奇形性情報コミュニテイの構築

Congenital malformations or birth defects are a major public concern. However, lack of proper information may panic a pregnant woman who had taken medicines without noticing the pregnancy, and thus may result in an unnecessary abortion. We have undertaken a new research/development project to create an information-community for teratogenic agents, consisting of pregnant women, pharmacists, physicians and research scientists working on medicinal products. We have designed a trial database system for teratogenic agents. The database performs not only retrieval of teratogenic information and chemical structures of drugs/medicinal products, but also registration of clinical case reports and teratogenic information on various chemicals. Our system will be useful for clinical risk-assessment of teratogenic exposures in pregnant women and for the development of new drugs based on chemical information and computer science.

薬物が原因で起こる胎児奇形は，頻度は低いが薬物治療において決して起こしてはならない副作用である．しかし，わが国には，妊娠に気づかずクスリを服用したとして過度の不安を抱いている妊婦に対する適切な情報提供システムがない．また，開発の中断された医薬品候補化合物の生殖・発生毒性情報は，公開され有効に利用されることなく企業の研究室に眠っているのが実状である．そこで，我々は，インターネット上に情報提供・収集システムを構築し，妊婦(市民)・医療関係者・研究者に情報コミュニティの場を提供することを目指している．先導研究として，Web上で医薬品催奇形性情報と化学構造式に関する条件検索が可能で，さらにクライアント側からも臨床症例登録のできる双方向・成長型の試用データベースを作成した．薬物催奇形性情報コミュニティの実現は，妊婦に対する根拠に基づいた薬物治療の実践と，情報化学的解析手法を用いた効率的な医薬品研究開発に資するものと期待される．


Project of the database construction for teratogenic agents-construction of the growing information -community for teratogenic agents


Thymocyte apoptosis induced by phosphorylation of histones is associated with the change in chromatin structure to allow easy accessibility of DNase

The inhibitors of protein phosphatase such as calyculin A and okadaic acid induce the apoptotic cell death in rat thymocytes. To clarify the molecular mechanism of these inhibitor-induced apoptosis, the effect of calyculin A on DNA fragmentation in the isolated nuclei were studied. A significant increase in DNA fragmentation was observed in the nuclei prepared from the cells treated with calyculin A that caused histone hyperphosphorylation. No changes of the activities of caspase-8 and -3 were observed in the extract from the cells treated with calyculin A. The circular dichroism analysis of soluble chromatin from calyculin A-treated thymocyte nuclei indicated that phosphorylation of histones decreased its alpha-helical content. Thus, the change in the chromatin structure may be due to the chemical modification of histones. Moreover, the structural change in chromatin preceded DNA fragmentation in the nuclei. Therefore, these results suggest that the change of chromatin structure allow easy accessibility of nuclear DNase to chromosomal DNA.


New entry of coupling reaction of phenacylamine derivatives with silylstannane

The new coupling reaction of phenacylamines with silylstannane and lithium diisopropylamide (LDA) is reported. The treatment of a phenacylamine iodide 1 with (trimethylsilyl)tributylstannane (Me_3SiSnBu_3) and cesium fluoride (CsF) gave a dimerization product 2 having no iodine atom. Reaction of 1 with LDA afforded a dimerization product 3 with an iodine atom. The products 2 and 3 were separated to the meso and racemic isomers, respectively.


Tissue Distribution of and Species Differences in Deacetylation of N-Acetyl-L-Cysteine and Immunohistochemical Localisation of Acylase I in the Primate Kidney

Species differences in the biotransformation of N-acetyl-L-cysteine (NAC) have been investigated to evaluate the usefulness of NAC as a constituent in parenteral nutrition solutions in place of cysteine. The activity of NAC-deacetylating enzyme (acylase) was measured in various tissues of different species (rat, rabbit, dog, monkey, and man). Acylase activity was highest in the kidney in all species studied. Enzyme activity in the liver was 10 %-22 % of that in the kidney in the rat, rabbit, monkey, and man, but almost no hepatic activity was seen in the dog. NAC-deacetylating activity was very low in other organs. The tissue distribution of acylase I was determined by Western blotting and an immunohistochemical method employing specific antibody against porcine acylase I (EC 3.5.1.14). The immunoblotting study showed a 46-kDa protein band corresponding to porcine acylase I in the kidney of all species. In liver cytosol, 46 kDa and/or 29 kDa bands were observed in the rat, rabbit, monkey, and man, but not in the dog. In the immunohistochemical study, positive staining with anti-acylase I antibody was observed clearly in the renal proximal tubules in the monkey and man. These results suggested that the kidney and liver were the main organs responsible for the biotransformation of NAC to cysteine in mammals other than the dog.

The Journal of Pharmacy and Pharmacology


Biotransformation of N-acetyl-L-cysteine:Tissue distribution and species difference of its deacetylation


Involvement of histone phosphorylation in thymocyte apoptosis by protein phosphatase inhibitors


Inhibitory effects of a cyclosporin derivative, SDZ PSC 833, on transport of doxorubicin and vinblastine via human p-glycoprotein

The inhibitory effects of SDZ PSC 833 (PSC833), a non-immunosuppressive cyclosporin derivative, on the P-glycoprotein (P-gp)-mediated transport of doxorubicin and vinblastine were compared with those of cyclosporin A (Cs-A). The transcellular transport of the anticancer drugs and PSC833 across a monolayer of LLC-GA5-COL150 cells, which overexpress human P-gp, was measured. Both PSC833 and Cs-A inhibited P-gp-mediated transport of doxorubicin and vinblastine in a concentration-dependent manner and increased the intracellular accumulation of doxorubicin and vinblastine in LLC-GA5-COL150 cells. The values of the 50%-inhibitory concentration (IC50) of PSC833 and Cs-A for doxorubicin transport were 0.29 and 3.66 microM, respectively, and those for vinblastine transport were 1.06 and 5.10 microM, respectively. The IC50 of PSC833 for doxorubicin transport was about 4-fold less than that for vinblastine transport, suggesting that the combination of PSC833 and doxorubicin might be effective. PSC833 itself was not transported by P-gp and had higher lipophilicity than Cs-A. These results indicated that the inhibitory effect of PSC833 on P-gp-mediated transport was 5- to 10-fold more potent than that of Cs-A, and this higher inhibitory effect of PSC833 may be related to the absence of PSC833 transport by P-gp and to the higher lipophilicity of PSC833.


Synthesis and biological evaluation of 7-hydroxy-3,4-diphenyl-1,2-dihydroquinolines as new 4-hydroxytamoxifen analogues


Involvement of histone hyperacetylation in triggering DNA fragmentation of rat thymocytes undergoing apoptosis

The treatment of rat thymocytes with trichostatin A and sodium butyrate, which are inhibitors of histone deacetylase, resulted in an increase in DNA fragmentation in a concentration-dependent manner. A significant increase in DNA fragmentation induced by these compounds was observed after a lag time of 2 h. Analysis of the fragmented DNA revealed the production of approximately 50 kb DNA fragments and DNA ladders, the biochemical hallmarks of apoptotic cell death. Judging from a laser scanning microscopic analysis, the inhibitors of histone deacetylase induced nuclear condensation, the morphological feature of apoptosis. Biochemical and morphological analyses demonstrated that trichostatin A and sodium butyrate induced thymocyte apoptosis. Furthermore, hyperacetylation of nuclear histones was observed in thymocytes treated with the inhibitors of histone deacetylase. These effects of sodium butyrate and trichostatin A were seen 0.5 and 1 h, respectively, after incubation of the cells. These results thus indicate that hyperacetylation of nucleosomal histones precedes DNA fragmentation in thymocytes undergoing apoptosis induced by trichostatin A and sodium butyrate.


Effect of Food Additives (Natural Red Colors) on Rabbit Platelet Functions.

食品添加物天然赤色系着色料のウサギ血小板機能に及ぼす影響

Japanese Journal of Toxicology and Environmental Health


Analysis of Tire Rubber Particles and Rubber Additives in Airborne Particulates at a Roadside.

道路近傍における浮遊粒子状物質中のタイヤレッド磨耗粉じん及びゴム添加剤の分析


Mutagenicity and property of airborne particulates in a roadside atmosphere

Japanese Journal of Toxicology and Environmental Health


Effect of food additives on rabbit platelet function ( II )


道路近傍における浮遊粒子状物質の変異原性について

道路近傍における浮遊粒子状物質の変異原性について


Food additives on acceptable daily intake (ADI) level affect the agonist induced platelet activation I. antioxidants and preservatives


食品添加物(赤色系合成着色料)のウサギ血小板機能に及ぼす影響

食品添加物(赤色系合成着色料)のウサギ血小板機能に及ぼす影響


Effect of food additives on rabbit platelet function


食品添加物(酸化防止剤)のウサギ血小板機能に及ぼす影響

食品添加物(酸化防止剤)のウサギ血小板機能に及ぼす影響


Purification of glutamine transferase K/cysteine conjugate β-lyase from rat renal cytosol based on hydrophobic interaction HPLC and gelpermeation FPLC

Cysteine conjugate beta-lyase (beta-lyase, EC 4.4.1.13) was purified to homogeneity from rat renal cytosol using a new and highly efficient method, based on C3-hydrophobic interaction (HI) high-performance liquid chromatography (HPLC) in combination with gel permeation fast protein liquid chromatography. The purity of the enzyme was judged from SDS-PAGE and C18-reversed-phase HPLC. The beta-lyase was estimated to be a homodimer consisting of a 47,400-Da subunit with absorption maxima at 280 and 420-430 nm. The specific activity of the purified beta-lyase toward S-(1,2-dichlorovinyl)-L-cysteine (1,2-DCVC) in the presence of alpha-keto-gamma-methiolbutyric acid (KMB) was 6.4 mumol/min/mg protein, which is by far the highest value so far reported. Kinetic analysis of 1,2-DCVC metabolism by the enzyme in the presence of KMB gave Km and Vmax values of 0.33 mM and 8.4 mumol/min/mg protein, respectively. No significant activity of the purified enzyme was detectable with S-2-benzothiazolyl-L-cysteine up to 2 mM. The purified enzyme also had glutamine transaminase K activity (EC 2.6.1.64) as assayed with phenylalanine and KMB as substrates. This specific activity was 16.0 mumol/min/mg. Amino acid analysis of the purified beta-lyase was carried out and was found to be closely similar to the amino acid composition of five other pyridoxal phosphate (PLP)-containing amino acid amino-transferases. This suggests that glutamine transaminase K/cysteine conjugate beta-lyase is a typical member of the PLP-containing aminotransferase group.


Effect of environmental pollutants on activation of rabbit platelets


Effect of halocarbons and styrene on thrombin and calcium ionophore induced activation of rabbit platelets


タイヤ抽出物の変異原性に対するゴム添加剤の共存効果

タイヤ抽出物の変異原性に対するゴム添加剤の共存効果


Rubber additive inhibit the calcium ionophore and thrombin induced activation of rabbit platelets


Effect of rubber additives on activation of rabbit platelets


Changes in the composition of glutathione transferase subunits in rat liver after oral administration of benzo(a)pyrene

Toxicological and Environmental Chemistry


タイヤ中に使用されているゴム添加剤の変異原性について

タイヤ中に使用されているゴム添加剤の変異原性について


水溶性ビタミン安定性に及ぼす赤色系着色料の影響

水溶性ビタミン安定性に及ぼす赤色系着色料の影響


Polycyclic aromatic hydrocarbons affect the calcium ionophore induced activation of rabbit platelet


Regulation of microsomal glutathione transferase activity in benzo(a)pyrene administered rat liver


High-performance liquid chromato-graphic analysis of polycyclic aromatic hydrocarbons extracted from river waters

Japanese Journal of Toxicology and Environmental Health


ブルーコットン吸着法による河川水中の多環芳香族炭化水素の分析

ブルーコットン吸着法による河川水中の多環芳香族炭化水素の分析


酸敗油脂水抽出-TBA比色法における反応生成物質の分析

酸敗油脂水抽出-TBA比色法における反応生成物質の分析


Relationship between central kinin system and hypertensive response in spontaneously hypertensive rat


Sex-related difference of kinin level in rat pituitary gland

The kinin level in the pituitary glands was compared in adult male and female rats. A sex-related difference in the bradykinin (BK)-like immunoreactivity was found in the posterior lobe. The posterior pituitaries of female rats contained a higher concentration of the immunoreactive kinin than those of males. Ovariectomy of female rats resulted in the disappearance of a sex difference in the posterior kinin level and about a 3-fold increase in the anterior one. Orchidectomy of adult male rats failed to alter the kinin levels in both lobes. Moreover, the constitution of pituitary kinins was determined using HPLC. The pituitary kinins consisted of BK, Lys-BK (L-BK) and Met-Lys-BK (ML-BK) in different proportions in both lobes of male and female rats. The gonadectomy altered the proportions of these kinins. These results suggest that the pituitary kinin system may be regulated by circulating gonadal steroid hormones.


Reactivity to bradykinin of the isolated ileum and duodenum from spontaneously hypertensive rats


Relationship between hypertensive response and brain kinin level in the rat injected intraventricularly with glandular kallikrein


A Study on kinin system in rat pituitary gland


Determination of kinin in the rat brain by a sensitive radioimmunoassay


Regional distribution and characterization of kinin in the CNS of the rat

The distribution of kinin in the CNS of the rat, which was extracted with n-butanol from an acidified homogenate, was determined using a bradykinin (BK) radioimmunoassay system. The immunoreactive kinin was widely distributed throughout the brain. The highest content was found in the pituitary gland (4,135 fmol BK Eq/g), followed by the medulla oblongata (912 fmol/g), cerebellum (549 fmol/g), and cortex (512 fmol/g). The kinin in the posterior pituitary was concentrated 4.5 times as much as in the anterior lobe. Serial dilution of brain extracts produced binding curves parallel to the standard radioimmunoassay curve. The purified brain kinin comigrated with authentic BK during CM-cellulose chromatography and Sephadex LH-20 gel chromatography. Its molecular weight was estimated to be 1,127 +/- 45 by gel filtration, which coincides well with that of BK. Chymotrypsin degraded the extracted kinin and authentic BK, but trypsin did not. These data demonstrate that a peptide indistinguishable from BK exists in the rat brain. Furthermore, pituitary kinin was separated into BK (87%), Lys-BK (10%), and Met-Lys-BK (3%), using reverse phase HPLC.


ラット下垂体キニン系に関する研究

ラット下垂体キニン系に関する研究


The disappearance rate of intraventricular bradykinin

Biochemical and Biophysical Research Communications


Relationship between central actions of bradykinin and prostaglandins in the conscious rat

Intraventricular injection of bradykinin produced a dose-dependent increase in the mean arterial blood pressure of conscious rats. With 5 nmol of bradykinin, a dual pressor response was observed, which was associated with a biphasic behavioral change. With repeated hourly injections of bradykinin, tachyphylaxis developed to the pressor and central nervous system (CNS) stimulating effect. Indomethacin, given intraventricularly, reduced the hypertension and the behavioral excitation caused by bradykinin in a dose-dependent manner. When prostaglandin E2 was injected into the cerebral ventricles, it induced hypertension and behavioral sedation similar to the secondary response to bradykinin. These results suggest that bradykinin has a dual action on the CNS, and this is mediated by prostaglandin-related systems in the brain.


Effects of intraventricular injection of bradykinin on the EEG and the blood pressure in conscious rats


Central action of bradykinin (II) Separation of bradykinin degrading enzyme from the rat brain


Effects of oral administration of positively charged insulin liposomes on alloxan diabetic rats

Insulin encapsulated in liposomes of various lipid compositions were prepared. The amount of insulin trapped in these liposomes increased in the order, negatively charged liposomes less than neutral liposomes less than positively charged liposomes. In positively charged liposomes, the amount of insulin trapped increased with increase in the amount of amphiphile stearylamine. Under the conditions tested, the highest insulin content (about 50%) was obtained with liposomes composed of phosphatidyl choline/cholesterol/stearylamine in a molar ratio of 7/2/2.25. These liposomes were stable on incubation for 3 hr at 37 degrees C in solutions of pepsin, trypsin, and pancreatin, and after these incubations, a considerable amount of insulin was still associated with the liposomes. However, the liposomes released almost all the insulin into the medium on treatment with bile. When the liposomes were administered orally to rats in the 3rd phase of acute alloxan diabetes, reduction of the blood glucose level was observed in 7 of 11 animals, the reduction persisted for several hours and was ranging from 30 to 75%. In alloxan diabetic rats showing hyperglycemia for 3 to 6 months, the liposomes also increased the glucose tolerance in half the animals tested.


Effect of bradykinin fragments and their analogs on central nervous system


Studies of the effects of bradykinin and its fragments on the central nervous system

Advances in Experimental Medicine and Biology


Development of thiol-responsive traceable linker for efficient enrichment and selective labeling of target proteins


Constructing a web-based drug-teratogenicity information sharing system, anzen-drug com


Estimating potential risks of pharmaceuticals human fetuses by SimScore in a teratogenicity information sharing system


セフェム系抗生物質のラット脳脊髄腔液移行性に関するQSAR解析

セフェム系抗生物質のラット脳脊髄腔液移行性に関するQSAR解析


薬物の胎盤通過性と母乳移行性予測-Clinical QSAR

薬物の胎盤通過性と母乳移行性予測-Clinical QSAR