Massage therapy for home care patients using the health insurance system in Japan.

OBJECTIVES: To clarify the status of home care massage services provided to patients. This will help in understanding how many patients utilize this service and the circumstances under which treatment is provided. DESIGN: A retrospective study. SETTING: Fifty-four acupuncture, moxibustion, and massage clinics. Participants were patients who had received home care massage for six months or more. We collected a total of 1587 responses from these 54 massage clinics; of these, 1415 responses (mean age = 79.1 ±â€¯11.5 years) were valid (valid response rate 89.2%). MAIN OUTCOME MEASURES: Actual patients and actual care services. RESULTS: The most common disorder observed among patients who utilized home care massage services was cerebrovascular disease (at approximately 36%), while the second most common were arthropathy-related disorders (16.3%). Although most patients received massage, approximately 30% received manual therapy (e.g. manual correction) and hot fomentation as part of thermotherapy. Notably, only around 10% of patients received massage alone; the majority received treatment in combination with range of motion and muscle-strengthening exercises. CONCLUSIONS: This study helped to clarify the actual state of patients receiving home care massage and the details of the massage services provided. This study clearly showed the treatment effectiveness of massage, which can be used by home medical care stakeholders to develop more effective interventions.

Structure and expression of the glycine cleavage system in rat central nervous system.

The glycine cleavage system (GCS) is a mitochondrial multienzyme system consisting of four individual proteins, three specific components (P-, T-, and H-proteins) and one house-keeping enzyme, dihydrolipoamide dehydrogenase. Inherited deficiency of the GCS causes nonketotic hyperglycinemia (NKH), an inborn error of glycine metabolism. NKH is characterized by massive accumulation of glycine in serum and cerebrospinal fluids and severe neuronal dysfunction in neonates. To elucidate the neuropathogenesis of NKH, we cloned cDNAs encoding three specific components of the GCS and studied the gene expression in rat central nervous system. P-, T-, and H-protein cDNAs encoded 1024, 403, and 170 amino acids, respectively. In situ hybridization analysis revealed that P-protein mRNA was expressed mainly in glial-like cells, including Bergmann glias in the cerebellum, while T- and H-protein mRNAs were detected in both glial-like cells and neurons. T- and H-protein mRNAs, but not P-protein mRNA, were expressed in the spinal cord. Primary astrocyte cultures established from cerebral cortex had higher GCS activities than hepatocytes whereas those from spinal cord expressed only H-protein mRNA and had no enzymatic activity. An important role of glycine as inhibitory neurotransmitter has been established in the brainstem and spinal cord and another role of glycine as an excitation modulator of N-methyl-D-aspartate receptor is suggested in the hippocampus, cerebral cortex, olfactory bulbus, and cerebellum. Our results suggest that the GCS plays a major role in the forebrain and cerebellum rather than in the spinal cord, and that N-methyl-D-aspartate receptor may participate in neuropathogenesis of NKH.

Drug design, synthesis, and evaluation of a non-sugar-based selectin antagonist.

We have designed a series of simple rigid compounds (2) having a phenyl ring attached to three essential groups necessary for selectin binding, i.e., a fucose unit, a carboxylic acid, and the hydrophobic part. In this series of compound 2, 2a exhibited strong inhibitory activity in in vitro P-selectin mediated cell adhesion assay. The novel type of compound 2a would be a potential lead compound for selectin antagonist.

Studies on selectin blocker. 9. SARs of non-sugar selectin blocker against E-, P-, L-selectin bindings.

As a part of study of selectin blockers, we have already reported that a non-sugar selectin antagonist (3) was successfully discovered using a computational screening (Hiramatsu, Y.; Tsukida, T.; Nakai, Y.; Inoue, Y.; Kondo, H. J. Med. Chem. 2000, 43, 1476). To investigate the SARs of compound 3 against E-, P-, and L-selectins, we synthesized the derivatives of compound 3 and evaluated their inhibitory activities toward selectin bindings. The structural diversity of compound 3 contained the following: (1) a modification of the spacer unit (4--7), (2) a modification of the tail unit (8--11), (3) a modification of the head unit (12--18). As a result, it was found that a non-sugar based selectin blocker (3) could be a potential lead compound for E-, P-, and L-selectin blockers and some of the derivatives showed broad and/or selective inhibitory activities toward the E-, P-, and L-selectins. In addition, it was found that the experimental evidence well supported that the computational screening using 3D-pharmacophore model could be useful methodology to find out a new lead for the several type of selectin blockers, which included a broad and/or a selective inhibitor.

Reversible thermal transition of soluble branched chains from slightly acid-treated potato starch.

The reversible thermal transition of soluble branched starch chains prepared from slightly acid-treated potato starch granules (ATS) was investigated. Potato starch was immersed in 15% sulfuric acid to obtain ATS with a 1% hydrolysis rate. About half of the molecules of ATS, which spontaneously formed large aggregates with a mass of a few million daltons in aqueous solution, was fractionated and soluble branched starch chains with a relative molecular weight (Mr) of 8.91 x 10(4) were obtained. Structural analysis indicated that the soluble branched starch chains consisted of three unit chains with Mr 7,900 and 21 unit chains with Mr 2,700. DSC and FT-IR measurements showed that the soluble branched starch chains underwent a reversible thermal transition, which is considered to be a helix-coil transition, during heating and cooling, but a debranched sample and beta-limit dextrins showed substantially different thermal behavior, indicating the contribution of the ordered structure of the branched chains.

Fertilizability and developmental capacity of individually cultured bovine oocytes.

Culture of single oocytes throughout in vitro maturation (IVM), fertilization (IVF) and culture (IVC) provides detailed information on maturity, fertilizability and developmental capacity of individual bovine oocytes and embryos. In the present study, effects of sperm concentration (Experiment 1), microdrop size (Experiment 2), and the addition of hypotaurine (HT) or glutathione (GSH; Experiment 3) during IVF were investigated. In Experiment 4, in vitro maturity and developmental capacity of bovine oocytes cultured for IVM in a medium supplemented with fetal calf serum (FCS), bovine serum albumin (BSA) or polyvinyl alcohol (PVA) during IVM were investigated. In Experiments 1 to 3, the percentages of normal (2 pronuclei with a spermtail) and polyspermic fertilization in singly cultured oocytes were similar to those of group IVF culture (5 oocytes/drop). The addition of GSH during single oocyte IVF significantly increased the proportion of normal fertilization and decreased the polyspermic fertilization compared with addition of HT or of the control. The rates of mature oocytes (62.4 and 67.7%) and blastocyst development (12.9 and 15.2%) for single oocyte IVM cultures (Experiment 4) were also similar compared with the group culture; PVA supplementation significantly increased the matured oocyte rate, but decreased blastocyst development significantly (7.1%) as compared with FCS (19.5%) or BSA (15.6%). These results indicate that a single oocyte culture system throughout in vitro production of bovine embryos provides similar maturity, fertilizability and developmental capacity to oocytes cultured in groups.

DNA sensing on a DNA probe-modified electrode using ferrocenylnaphthalene diimide as the electrochemically active ligand.

Naphthalene diimide derivative 1 carrying ferrocenyl moieties at the termini of imide substituents binds intact calf thymus DNA 4 times more strongly than the denatured DNA, and its complex with the intact DNA dissociates 80 times more slowly than that with the denatured DNA. On the basis of these observations, ligand 1 was applied to a probe of electrochemical DNA sensing. A thiol-linked single-stranded DNA probe was immobilized through the S-Au bonding to 20-30 pmol/mm2 on a gold electrode. Following hybridization with the complementary DNA, the electrode was soaked in a solution containing 1 (intercalation step) and then washed with buffer for 5 s. The cyclic voltammogram and differential pulse voltammogram for this electrode gave an electrochemical signal due to the redox reaction of 1 that was bound to the double-stranded DNA on the electrode. Thus, dA20 and the yeast choline transport gene were quantitated at the subpicomole level. The sensitivity of DNA detection was improved to 10 zmol by reducing the amount of immobilized DNA probe and protecting the uncovered surface of the electrode with 2-mercaptoethanol.

Distribution and intracellular localization of a mouse homologue of Ca2+/calmodulin-dependent protein kinase Ibeta2 in the nervous system.

Ca2+/calmodulin-dependent protein kinases (CaMKs) are believed to play important roles in the development and function of the nervous system. We report here the identification and expression of mouse CaMKIbeta (mCaMKIbeta), in particular mCaMKIbeta2, an isoform of mCaMKIbeta. During embryogenesis, the mCaMKIbeta2 gene is expressed mainly in the nervous system, including brain, spinal cord, trigeminal ganglion, and retina. Within the CNS, the expression of mCaMKIbeta2 is detected in the mantle zone, but not in the ventricular zone, suggesting its possible involvement in the differentiation of neurons. In the adult brain, mCaMKIbeta2 transcripts are detected at high levels in the anterior olfactory nuclei, piriform cortex, septal nuclei, bed nuclei of the stria terminalis, hippocampal pyramidal cells, dentate granule cells, amygdala, hypothalamic nuclei, parabrachial nucleus, and nucleus of the solitary tract. The distinct gene expression pattern suggests that mCaMKIbeta2 may also be involved in different mature neuronal functions from other CaMKs. In addition, mCaMKI/beta2 proteins are localized to the cytoplasm and nuclei, but not to nucleoli, suggesting that mCaMKIbeta2 proteins might be involved in the cytoplasmic and nuclear signal transduction of the nervous system.

Attenuation of monosodium urate crystal-induced arthritis in rabbits by a neutralizing antibody against interleukin-8.

Accumulating evidence implicates interleukin-8 (IL-8) as an essential mediator in neutrophil-mediated acute inflammation. Neutrophils have also been shown to have a crucial role in the pathogenesis of acute gouty arthritis. Thus, we investigate the pathophysiological role of IL-8 in an experimental model of acute gout, monosodium urate (MSU) crystal-induced arthritis in rabbits. The injection of MSU crystals into knee joints caused a marked swelling of joints. Concomitantly, the infiltration ofleukocytes, mostly neutrophils, was observed in synovial membrane and synovial fluids. The injection of MSU crystals also induced an elevation in synovial fluid IL-8 levels preceding neutrophil infiltration into synovial fluids, without an accompanying increase in plasma IL-8 levels. Immunoreactive IL-8 protein was detected in synovial lining cells at 12-24 h after the injection. IL-8 protein was also observed in infiltrated leukocytes in synovium as early as 3-24 h after the injection. Finally, the intraarticular injection of a neutralizing anti-IL-8 antibody significantly attenuated the crystal-induced joint swelling that occurred at 12 h, and neutrophil infiltration into arthritic joints at 12 and 24 h after the induction. These results provide evidence on the pathogenic roles of locally produced IL-8 in MSU crystal-induced gouty arthritis.

Cloning and expression localization of cDNA for rat homolog of TRP protein, a possible store-operated calcium (Ca2+) channel.

A 3.2 kbp cDNA clone encoding a possible candidate for the store-operated Ca2+ channel was isolated from a rat brain cDNA library. The deduced amino acid sequence was 51.8% identical to TRP encoded by a Drosophila trp (transient receptor potential) gene and contained ankyrin motifs, a coiled-coil structure and six transmembrane segments similar to the previously identified TRP family and named as TRP-R (rat TRP). By in situ hybridization histochemistry of rat body on embryonic day 15, no significant expression signal for TRP-R was detected. On embryonic day 20 and postnatal day 1, the expression signals were most evident in the septum, cerebral cortical plate and hippocampal neuronal layers. On postnatal day 7 and thereafter the expression in the cerebral cortex and the septum decreased progressively, and weak expression remained only in the CA1 and CA2 neuronal layers of the hippocampus in the brain on postnatal day 21 and 49. This limited spatiotemporal expression of this novel molecule. TRP-R, suggests that it is involved in some specific functions related to the neuronal differentiation.

Influence of dietary fiber on the bioavailability of zinc in rats.

Young male albino rats were fed ad libitum semipurified diet supplemented with or without 5% of purified dietary fiber (cellulose, agar-agar, pectin, chitin or chitosan) for 31 days. Each test diet was carefully prepared in order to contain zinc at the level of 8 ppm from zinc acetate. In rats fed diets containing 5% of dietary fiber except chitosan, food consumption was higher than the control. The body weight gain of rats fed diets containing cellulose, pectin or chitin was higher than the control. However, food intake and body weight gain of rats fed the diet containing 5% of chitosan were definitely lower than not only the other fiber including diet groups but also the control group. When dietary fiber was added at 5% level to diet, zinc absorption was not changed to a considerable degree. But, the apparent zinc absorption of rats fed the agar-agar diet was 70%. On the other hand, zinc absorption in the pectin diet group was about 10% higher than the control. The total amount of zinc in tibia or femur of rats fed non-fiber diets was a little higher than that of rats fed fiber diets.

Localization of mRNAs of voltage-dependent Ca(2+)-channels: four subtypes of alpha 1- and beta-subunits in developing and mature rat brain.

The heterogeneous gene expression for four subtypes of alpha 1 (A,B,C,D)- and beta (beta 1,beta 2,beta 3,beta 4)-subunits of voltage-dependent calcium channels was demonstrated in developing and adult rat brain by in situ hybridization histochemistry. In the adult rat brain the gene expression for A- and B-subtypes was predominant in the cerebellar cortex and hippocampal neuronal layers, with the A-subtype expressed most intensely in the Purkinje cells, while the expression for C- and D-subtypes was predominant in the olfactory mitral and granule cells and the dentate granule cells. The expression of beta 1-mRNA was prominent in the olfactory mitral cells and dentate granule cells whereas that of beta 2-mRNA was evident in the hippocampal neuronal layers and cerebellar Purkinje cells. The expression of beta 3-mRNA was prominent in the olfactory mitral and internal granule cells and medial habenula, whereas that of beta 4-mRNA in the olfactory mitral cells and cerebellar Purkinje and granule cells. Comparison between the expression patterns for individual alpha- and beta-subunits suggests that the beta 4-subunit contributes to P-type channel, whereas the beta 1- and beta 3-subunits contribute respectively to D- and C-subtypes of L-type channels, although dissociation in the expression patterns were also noted in several brain regions. In addition to neuronal populations, the gene expression for the C-subtype of L-type channel was detected at substantial level in glial cells. In developing brains, the genes for the all subtypes of alpha 1- and beta-subunits were expressed in the mantle zones, but not the ventricular zones, of the entire neuraxis and the expression was more or less attenuated during early postnatal periods in most of the brain regions except for the olfactory bulb, hippocampus and cerebellar cortex, suggesting that the Ca(2+)-channels are intimately involved in the neuronal differentiation.

Cloning and expression of a cytoskeleton-associated diacylglycerol kinase that is dominantly expressed in cerebellum.

A third species of diacylglycerol kinase (EC 2.7.1.107) cDNA was cloned from a rat brain cDNA library. The isolated cDNA encoded a 788-amino acid, 88-kDa polypeptide. This isozyme shared 58% identity with the previously isolated rat 80-kDa and 90-kDa diacylglycerol kinases. EF hand motifs, cysteine-rich zinc finger-like sequences, and putative ATP-binding site were all conserved among these isozymes. The 88-kDa diacylglycerol kinase was expressed specifically in brain and localized predominantly in cerebellar Purkinje cells. This isozyme was associated equally with particulate and supernatant fractions in cDNA-transfected COS-7 cells and dominantly with the particulate fraction in the brain. After Triton X-100 extraction, this isozyme remained in the detergent-insoluble cytoskeletal fraction of the brain and transfected COS-7 cells.

Molecular cloning of rat cDNAs for the zeta and theta subtypes of 14-3-3 protein and differential distributions of their mRNAs in the brain.

We isolated from the rat brain two cDNA clones encoding the zeta and theta subtypes of the 14-3-3 protein. Both clones encoded 245 amino acid sequences, which share a high sequence homology with each other and also with other subtypes of the 14-3-3 protein. The distribution of their mRNAs was determined in the developing brain, by in situ hybridization with subtype-specific oligonucleotide probes. At embryonic day 18, the zeta and theta subtype mRNAs were expressed at high levels throughout the brain and the spinal cord. Distribution patterns of the two mRNAs were distinct in the brain at postnatal day 21. The zeta subtype mRNA was distributed widely in the brain gray matter, and high levels of the transcripts were detected in various brain regions, including the neocortex, hippocampus, caudate-putamen, thalamus, cerebellar cortex, and several brainstem nuclei. On the other hand, high signal levels of the theta subtype mRNA in the gray matter were restricted to the cerebellar cortex and the hippocampus. In addition, significant signals for the theta subtype mRNA were found over the white matter, where cell bodies of glial cells are populated. The wide gene expression of the zeta and theta subtypes suggests their fundamental and essential role in the brain function, but the degrees of functional involvement by the respective subtypes would be heterogeneous between neuron and glia, and also among neuron types.

Molecular cloning of a novel isotype of Mg(2+)-dependent protein phosphatase beta (type 2C beta) enriched in brain and heart.

Two complementary DNA (cDNA) clones (pTK-1 and -2) encoding two distinct isotypes of mouse Mg(2+)-dependent protein phosphatase beta (MPP beta-1 and -2, respectively) were isolated from a melanocyte cDNA library. Although mouse pTK-1 is orthologous to the rat cDNA (JW5) reported previously [Wenk, J., Trompeter, H.I., Pettrich, K.G., Cohen, P.T.W., Campbell, D.G., and Mieskes, G. (1992) FEBS Lett. 297, 135-138], pTK-2 is a novel cDNA clone. It was strongly suggested that the pTK-1 and -2 cDNAs are splicing variants of a single pre-mRNA. The difference in the amino acid sequences between MPP beta-1 and -2 was observed only at the carboxy-terminal regions. Both the recombinant MPP beta-1 and -2 expressed in Escherichia coli cells were immunoreactive to an anti-MPP beta antibody and exhibited Mg(2+)-dependent and okadaic acid-insensitive protein phosphatase activities with similar substrate specificities. Although the mRNA of MPP beta-1 was expressed ubiquitously in various mouse tissues, that of MPP beta-2 was expressed exclusively in brain and heart. These results suggest the difference in the physiological roles of these two enzyme isotypes.

[Megakaryoblastic leukemia which developed from therapy-related MDS with myelofibrosis].

A 56-year-old man had a leiomyosarcoma of the small intestine in 1987. After surgery, he received cyclophosphamide for 2 years. In December, 1990, he exhibited severe pancytopenia. His hematological data were as follows: Hb 7.4g/dl, ret. 0.8%, WBC 1,700/microliters with leukoerythroblastosis and 2.8 x 10(4)/microliters platelets. A bone marrow aspiration was a dry tap. A bone marrow biopsy specimen showed a hypercellular marrow with myelofibrosis, leukemic infiltration (10.2%) and slight dyserythropoiesis. Both PPO and GPIIb/IIIa reaction were positive for blast cells and atypical megakaryoblasts. A diagnosis of MDS with an abnormality in megakaryocytic lineage was made. The patient was treated with 1,25-dihydroxy-vitamin D3, however this therapy was temporary and he developed into acute megakaryoblastic leukemia (M7). This report suggested that some cases of therapy-related leukemia (TRL) mainly involve megakaryocytic lineage and are diagnosed as MDS with myelofibrosis which transform to M7. The fact that PAS stain of erythroblasts in the patient reported here was positive may suggest involvement of development of more precise immunological markers of differentiation and EM study will permit better diagnosis of TRL and may therefore facilitate new therapeutic approaches.

Inhibition of herpes simplex virus infection by pine cone antitumor substances.

Several antitumor substances extracted from cones of various pine trees inhibited the plaque formation of Herpes simplex virus types 1 and 2 (HSV-1, HSV-2) strains in African green monkey kidney cells and human adenocarcinoma cells. The 50% effective dose of the most active fraction, Fr. VI (0.3 micrograms/ml) was 0.001 times its 50% cytotoxic dose (greater than 300 micrograms/ml). The anti-HSV activity of various pine cone extracts increased with their acidity. Identification of the polyphenol groups as donors of the acidity was further supported by the observation that the anti-HSV activity of the natural polyphenolic products, such as tannin and lignin, significantly exceeded that of other natural or chemically modified antitumor polysaccharides. An experiment using radiolabeled virus particles indicated that the anti-HSV effect of both Fr. VI and lignin was attributable to interference with virus adsorption to these cells rather than to inhibition of virus penetration into the cells.

Cobalamin-dependent replication of L1210 leukemia cells and effects of cobalamin analogues.

L1210 cell line cells were made deficient in Cbl by propagation in a medium devoid of CNCbl in which fetal calf serum had been replaced by bovine serum albumin. These Cbl-deficient cells gradually ceased to multiply when the medium contained 5-CH3THF, although cell growth was resumed following the addition of CNCbl, OHCbl or folic acid. The results of this study provide experimental proof for the 'methyl trap' hypothesis. In contrast to the above effect of CNCbl, cobinamide and Cbl analogues which were produced by a reaction of OHCbl with ascorbic acid did not have any growth-inducing effect on the cells which had been cultured in a 5-CH3THF-supplemented medium and had ceased to multiply. Nor did these analogues have an inhibitory effect on CNCbl-dependent growth.

Serum transferrin receptor as a new index of erythropoiesis.

Serum transferrin receptors were measured by a sandwich radioimmunoassay procedure in patients with iron deficiency anemia, autoimmune hemolytic anemia and aplastic anemia. The mean circulating transferrin receptor concentration of normal subjects and patients with iron deficiency anemia, autoimmune hemolytic anemia and aplastic anemia are 253 +/- 82 ng/mL, 730 +/- 391 ng/mL, 1,426 +/- 1,079 ng/mL, and 182 +/- 39 ng/mL, respectively. The values for those with iron deficiency anemia and autoimmune hemolytic anemia were significantly higher than that of normal controls and the values for those with aplastic anemia were lower than that of normal controls. After iron supplementation in iron deficiency anemia, the serum transferrin receptor values increased twofold over those of pretreatment values. This increase parallels an increase in peripheral reticulocytes. Therefore, the number of circulating transferrin receptors in anemic patients may reflect the level of bone marrow erythropoiesis and is a potentially useful new index for red cell production.

On RNA-polymerases of leukemia L 1210 origin and an enzymatic method to screen antitumor antibiotics.

Four DNA-dependent RNA-polymerases were separated from the cell homogenate of moust leukemia L1210 cell by DEAE-cellulose column chromatography and tentatively designated as Peaks I, II, III and IV in the elution order. Peak II was inactivated by the addition of alpha-amanitin and effects of antibiotics and enzymes on the RNA-polymerase activity using Peaks, I, II and a mixture of Peaks I and II were examined. The RNA-polymerases were used to screen for enzyme inhibitors produced by microbes. This enzymatic method was successfully proved to select antitumor antibiotics.