The enhancing effect of a triethanolamine-ethanol-isopropyl myristate mixed system on the skin permeation of acidic drugs

The effect of a TEI enhancer mixed system consisting of triethanolamine (T), ethanol (E) and isopropyl myristate (IPM) on the skin permeation of acidic, basic and neutral drugs were evaluated in vitro using excised hairless rat skin. The binary enhancer system consisting of IPM and ethanol (El) produced marked improvement on the penetration of all the drugs tested. When T was added to the EI system, a greater enhancing effect was found only on acidic drugs with a carboxyl group, compared with the flux in the EI system. On addition of another amine to the EI system, instead of T, mefenamic acid (MA), which exhibited the highest enhancing effect of the model drugs, showed an approximately 14-180 times greater flux than when delivered by the EI system. On simultaneous application of isosorbide dinitrate (ISDN) with MA in the TEI system, the flux of MA increased on increasing the T concentration in the TEI system, while, the flux of ISDN, a neutral drug, was unaffected by the T concentration. Application of MA in the EI system after pretreatment of the TEI system showed that the residual amount of T in the skin plays an important role in the skin permeation of MA. Furthermore, at a fixed concentration of MA, the flux of MA increased on increasing the T concentration in the TEI system, while the flux of E remained unchanged. Finally, the infrared spectrum of MA with amine in the E solution indicated that the carboxyl group of MA was ionized. These results demonstrated that the formation of an ion pair between MA and T, but not the effect of T on the skin, may be responsible for the enhanced skin permeation of MA using the TEI system.     

Potential usage of liposomal 4beta-aminoalkyl-4'-O-demethyl-4-desoxypodophyllotoxin (TOP-53) for cancer chemotherapy

To enhance the therapeutic efficacy as well as to reduce the side effect, we attempted to liposomalize 4beta-aminoalkyl-4'-O-demethyl-4-desoxypodophyllotoxin (TOP-53), a novel and effective topoisomerase II inhibitor. More than 90% of TOP-53 was efficiently incorporated into the liposomes composed of dipalmitoylphosphatidylcholine and cholesterol by remote-loading method. Anti-tumor activity of liposomal TOP-53 against solid tumor was examined in vivo using colon26 NL-17 carcinoma model mice. Three doses of liposomal TOP-53 (12 mg/kg/dose) showed significant tumor growth suppression (97.5% reduction determined at day 25) and the increase in life span (33%) of tumor-bearing mice. Furthermore, one mouse out of 5 was completely cured after treatment. Since similar efficacy was observed in the free TOP-53 treated group, liposomalization does not contribute much to the enhancement of therapeutic efficacy. However, a slight but measurable damage at the injection site was observed when free TOP-53 was injected, and the damage was diminished by the liposomalization. Taken together, liposomalization reduces the side effect rather than enhancing the therapeutic efficacy when TOP-53 is used.     

Effective treatment of bladder tumor-bearing mice by direct delivery of bleomycin using electrochemotherapy

We investigated the cytotoxicity on the combination of bleomycin (BLM) with electric pulses against transplanted bladder tumors in nude mice. Loading with high-voltage electric pulse after injection of BLM was associated with a decrease in tumor size; tumor size became smaller and disappeared 8 days after the treatment. Such complete regression was achieved with one treatment using BLM of one-tenth the lethal dose (LD 50 ) combined with electric pulses. On day 20, regrowth was observed at a much slower rate than at lower concentrations of BLM. Pathological examinations of the tumor tissues after treatment revealed disruption of the nucleus in the exposed tissues on day 2. Furthermore, a decrease in number of the stained nuclei and cytoplasmic lysis were observed on day 4. These results showed that electric pulses was an effective tool to deliver drugs into bladder tumor cells and an effective treatment method for bladder tumors.     

Research on factors for dropout from treatment by outpatients of alcohol abuse or alcohol dependence (II)--Based on questionnaire's outcome]

This research examines a strategy to avoid dropout from treatment by outpatients of alcohol abuse and dependence. Questionnaire was used and outcome was analyzed in order to elucidate factors of dropout and examine measures for its avoidance. The questionnaire made on the occasion of "Research on Factors of Dropout from Treatment by Outpatients of Alcohol Abuse or Dependence (I)" was used. 376 subjects was surveyed with 178 answers (47.3%), out of which 167 (44.5%) were valid. The analyses of answers showed as factors of dropout: dissatisfaction for content and environment of treatment, transfer to internal medicine department, and almost every day drinking. Reasons for continuation of treatment were: satisfaction for content of medical examination, admission to specialized treatment facilities, participation to self-help groups, feeling of happiness for treatment, since beginning intended to continue treatment. As a measure to avoid treatment dropout, improvements of content and environment of treatment and better relationship among medical staffs and clients were suggested.     

Eleven novel single nucleotide polymorphisms in the NR1I2 (PXR) gene, four of which induce non-synonymous amino acid alterations

Eleven novel single nucleotide polymorphisms (SNPs) were found in the NR1I2 (PXR/SXR) gene from 205 Japanese subjects. The detected SNPs were as follows: 1) SNP, MPJ6_1I2001; GENE NAME, NR1I2; ACCESSION NUMBER, AF364606; LENGTH, 25 bases; 5'-TTTCTACCTCTAC/TTATTGAAAGGGC-3'. 2) SNP, MPJ6_1I2004; GENE NAME, NR1I2; ACCESSION NUMBER, AF364606; LENGTH, 25 bases; 5'-AGGCCCAAATGTG/AAGTGATGCATAG-3'. 3) SNP, MPJ6_1I2007; GENE NAME, NR1I2; ACCESSION NUMBER, AF364606; LENGTH, 25 bases; 5'-TGCCAGGCCTGCC/TGCCTGCGCAAGT-3'. 4) SNP, MPJ6_1I2008; GENE NAME, NR1I2; ACCESSION NUMBER, AF364606; LENGTH, 25 bases; 5'-GAGTGAGCAGTGG/CGCGCGCGGGCGG-3'. 5) SNP, MPJ6_1I2010; GENE NAME, NR1I2; ACCESSION NUMBER, AF364606; LENGTH, 25 bases; 5'-CAGAGGAGCAGCG/AGATGATGATCAG-3'. 6) SNP, MPJ6_1I2011; GENE NAME, NR1I2; ACCESSION NUMBER, AF364606; LENGTH, 25 bases; 5'-CTGGAAGTGGCCA/GGGAGGTTCAAAG-3'. 7) SNP, MPJ6_1I2013; GENE NAME, NR1I2; ACCESSION NUMBER, AF364606; LENGTH, 25 bases; 5'-TCTTCCTCTCGCC/TCCCAACTTCTGG-3'. 8) SNP, MPJ6_1I2017; GENE NAME, NR1I2; ACCESSION NUMBER, AF364606; LENGTH, 25 bases; 5'-ATTGAATGCAATC/TGGCCCCAGCCTG-3'. 9) SNP, MPJ6_1I2018; GENE NAME, NR1I2; ACCESSION NUMBER, AF364606; LENGTH, 25 bases; 5'-GGTGAGCACAGCA/GGGGGGTGAGGAC-3'. 10) SNP, MPJ6_1I2019; GENE NAME, NR1I2; ACCESSION NUMBER, AF364606; LENGTH, 25 bases; 5'-GAGCTCCGCAGCA/GTCAATGCTCAGC-3'. 11) SNP, MPJ6_1I2021; GENE NAME, NR1I2; ACCESSION NUMBER, AF364606; LENGTH, 25 bases; 5'-GGTGACACCTCCG/AAGAGGCAGCCAG-3'. The frequencies were 0.0293 for MPJ6_1I2021, 0.0073 for MPJ6_1I2011, and 0.0024 for the other 9 SNPs. All SNPs were found as heterozygous. Among these SNPs, MPJ6_1I2007, MPJ6_1I2010, MPJ6_1I2017 and MPJ6_1I2019 induce non-synonymous amino acid alterations (R98C, R148Q, R381W and I403V, respectively, in PAR1).     

A major genotype in UDP-glucuronosyltransferase 2B15

A novel single nucleotide polymorphism (SNP) was found in exon 6 of the UDP-glucuronosyltransferase (UGT) 2B15 gene from healthy Japanese populations. The SNP was as follows: SNP, 020228Toide001; GENE NAME, UGT2B15; ACCESSION NUMBER, U08854, AF180322, and NM_001078; LENGTH, 25 base; 5'-AGCTTGCCAAAAC/AAGGAAAGAAGAA-3'. This SNP was expected to cause a change of an amino acid residue at the position 523 (Thr to Lys) located in a putative co-factor binding region. The allele frequency of this SNP was 79% in Japanese, suggesting this polymorphism to be a major genotype in Japanese people.     

Chinese medicine induces neurite outgrowth in PC12 mutant cells incapable of differentiation

During continuous culture of neural PC12 cells, we obtained a drug-hypersensitive PC12 mutant cell that showed high stimulation of neurite outgrowth by various drugs. When several Chinese medicines such as shu-jing-huo-xie-tang and Wu-Ling-San were provided to these PC12 mutant cells, the frequency of nerve growth factor (NGF)-induced neurite outgrowth increased approximately 30-fold compared to NGF alone. Neurite outgrowth induced by NGF in PC12 cells is accompanied by sustained activation of mitogen-activated protein kinase (MAPK); however, these Chinese medicines did not induce MAPK activity. The findings thus indicate that certain Chinese medicines may induce neurite outgrowth by a novel mechanism which is distinct from the NGF-activated pathway in PC12 mutant cells.     

Functional anatomy of the tensor veli palatini muscle and Ostmann's fatty tissue

This study examined morphological features of the tensor veli palatini muscle (TVPM) and Ostmann's fatty tissue that may be important for eustachian tube (ET) ventilation. Histologic sections through the midcartilaginous ET from 17 human temporal bone-ET specimens (age range. 3 months to 88 years) were used to assess 1) the presence or absence of attachment of the TVPM fibers to either the perichondrium of the ET cartilage lateral lamina (LL) or a tendinous membrane along the medial margin of the TVPM, 2) the angular relationship between the TVPM fibers and the vertical axis of the ET lumen, and 3) the location of the TVPM and Ostmann's fatty tissue. The TVPM fibers were attached to the LL perichondrium in 14 cases; an attachment was absent in 3 cases because of fatty atrophy of the TVPM. However, the TVPM fibers were inserted into the tendonlike membrane in all cases. The angle of insertion of TVPM fibers into the membrane was significantly more acute (relative to the vertical ET axis) in the inferior aspect than in the superior aspect of the membrane both in young children (3 months to 4 years; mean +/- SD, 39.0 degrees +/- 15.1 degrees superiorly to 23.8 degrees +/- 17.0 degrees inferiorly) and in older subjects (8 to 88 years, 30.4 degrees +/- 11.6 degrees superiorly to 15.7 degrees +/- 11.2 degrees inferiorly; t-test, p < .001).The location of Ostmann's fatty tissue accompanied the TVPM throughout the cartilaginous ET. These data suggest that contraction of the TVPM moves the LL inferolaterally to open the superior aspect more than the inferior aspect of the lumen and that Ostmann's fatty tissue will limit the opening of the ET lumen, especially that of its inferior aspect.