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Biomedical subjects

K Nakashima

Publications and source records attributed to K Nakashima.

At least 37 records · Page 2Linked to original sources
PubMed2003

Bone morphogenetic protein-2 enhances osterix gene expression in chondrocytes.

Osterix is a recently identified zinc-finger-containing transcription factor, which is required for skeletogenesis as no bone formation was observed in osterix-deficient mice. Osterix was first cloned as a gene whose expression was enhanced by BMP in C2C12 cells. As BMP induces ectopic bone formation in vivo via a pathway reminiscent to endochondral bone formation, BMP may also regulate osterix gene expression in chondrocytes. However, no information was available regarding the BMP actions on osterix gene expression in chondrocytes. We therefore examined the effects of BMP-2 on osterix gene expression in chondrocytes in culture. RT-PCR analysis indicated that osterix mRNA was expressed in the primary cultures of chondrocytes derived from mouse rib cartilage. The treatment with BMP-2 enhanced the levels of osterix transcripts within 24 h and the enhancement was still observed at 48 h based on RT-PCR analysis. This BMP effect was specific to this cytokine, as TGF-beta did not alter osterix gene expression. BMP effects on the osterix mRNA levels were also confirmed by Northern blot analysis. The enhancing effect of BMP on osterix gene expression was observed in a dose-dependent manner starting at 200 ng/ml. The BMP enhancement of the osterix gene expression in chondrocytes was blocked in the presence of a protein synthesis inhibitor, cycloheximide, while it was still observed in the presence of 5,6-dichloro-1-beta D-ribofuranosylbenzimidazol (DRB) suggesting the involvement of post-transcriptional events, which require new protein synthesis. These results indicated that osterix gene is expressed in the primary cultures of chondrocytes and its expression is under the control of BMP-2.

Animals↗
PubMed2003

Production of dopaminergic neurons for cell therapy in the treatment of Parkinson's disease.

Dopaminergic cell therapy is a potential viable treatment for Parkinson's disease. However, lack of a well-characterized cell preparation of known phenotypic composition containing a high percentage of dopaminergic neurons, has prevented a definitive, controlled, pilot clinical trial from being conducted. We report the successful in vitro expansion of rat E12 mesencephalic progenitors to produce 5-fold the normal number of dopaminergic neurons. The expanded neurons (MAP2+) were detached, resuspended, and formed into small aggregates of 10-200 neurons containing 25-50% of dopaminergic neurons (TH+) that will likely be optimal for use in successful cell therapy. After storage in DPBS, in 0 mM Ca(2+) for up to 24 h at room temperature, aggregated cells were still 90% viable. These results demonstrate that it might be feasible to use a similar protocol to expand human dopaminergic progenitors in vitro. If successful, the requisite large numbers of dopaminergic neurons required to conduct a pilot clinical trial for Parkinson's disease will be produced in vitro. Indications are that the cells can be maintained at optimal viability for the duration of the neural transplantation procedure, under real operating conditions.

Animals↗
PubMed2003

Axonal and demyelinating forms of the MPZ Thr124Met mutation.

OBJECTIVE: We report on a Japanese family with Charcot Marie Tooth disease (CMT) with the Thr124Met mutation in the peripheral myelin protein zero (MPZ) gene. MATERIAL AND METHODS: Based on the clinical study, we investigated MPZ gene by direct sequence analysis and polymerase chain reaction restriction fragment length polymorphism analysis. RESULTS: Genotyping of four symptomatic family members showed that one family member with severe disease symptoms was homozygous, while the other three were heterozygous. The heterozygous cases were clinicopathologically determined to be the axonal type, which is characterized by late-onset and slow progression associated with Adie's pupil and deafness. The homozygous case was the demyelinating type, which showed earlier onset, rapid progression, sural nerve demyelination, and cranial nerve demyelination at autopsy. CONCLUSIONS: We suggest that axonal and demyelinating forms of CMT are not two distinct classes, but rather parts of a spectrum of genotypically related conditions, particularly with some MPZ mutations.

Adult↗
PubMed2003

NRAMP1 polymorphisms, susceptibility and clinical features of tuberculosis.

OBJECTIVES: To study whether NRAMP1 gene polymorphisms in a Japanese population affect susceptibility to tuberculosis and clinical features of tuberculosis. METHODS: Polymerase chain reaction and restriction fragment-length polymorphism analyses were used to type NRAMP1 polymorphisms in 95 patients with pulmonary tuberculosis and 90 controls. Clinical features of patients also were investigated. RESULTS: No association was seen between susceptibility to tuberculosis and NRAMP1 polymorphisms. Patients with the D543N A allele were significantly more likely than others to develop a cavitary lesion; by logistic regression analysis with adjustment for gender, age, and presence of diabetes, the odds ratio in patients with an A allele was 5.16 (95% confidence interval, 1.30-20.45). CONCLUSIONS: Genetic variation in the human NRAMP1 gene may be associated with cavitation in patients with tuberculosis.

Adult↗
PubMed2003

Glutathione S-transferase polymorphisms: susceptibility to migraine without aura.

Migraine is considered to be a polygenic multifactorial disease with various environmental and genetic etiologies. We investigated glutathione S-transferase (GST) P1 Ile(105)Val, T1 and M1 polymorphisms in 174 Japanese headache sufferers and 372 Japanese controls. The headache group consisted of 38 cases of migraine with aura, 95 migraine without aura (MWOA) and 41 tension-type headache sufferers. The M1 homozygous deletion genotype was significantly higher in MWOA (64%) compared with controls (46%; p < 0.01; odds ratio = 2.18, 95% confidence interval: 1.32-3.61, adjusted for age and gender). In a comparison of the current smokers, the M1 null frequencies in MWOA were further increased. GSTM1 may be one of the genetic risk factors for MWOA in the Japanese population.

Adult↗
PubMed2003

[The isolated congenital mitral regurgitation due to a partial defect of the anterior leaflet].

The isolated congenital mitral regurgitation (MR) is a rare anomaly. We have successfully treated a 7-year-old boy who had MR due to a partial defect of the anterior leaflet. Echocardiogram revealed moderate MR at the anterior leaflet as a result of its partial defect. Another MR was also detected at the coaptation zone of the 2 leaflets due to mild prolapse of the anterior leaflet. The mitral valve was approached via atrial septostomy and a partial defect (8 x 6 mm) was found near the postero-medial commissure of the anterior leaflet. Any chordae did not develop around the defect or subvalvular lesions. Other parts of the leaflet was seemed normal and any inflammatory or degenerative changes were not detected in the rest of the leaflets. We have done direct closure of the defect of the leaflet and also added annuloplasty at the posteromedial commissure by modified Kay method.

Cardiac Surgical Procedures↗
PubMed2002

Three-dimensional fast recovery fast spin-echo imaging of the inner ear and the vestibulocochlear nerve.

The aim of this study was to assess the performance of three-dimensional fast recovery fast spin-echo (3DFRFSE) for imaging of the inner ear as well as the facial and vestibulocochlear nerves. We evaluated 3DFRFSE sequences, comparing it with 3D fast spin-echo (3DFSE) in a water phantom and in 12 normal volunteers. We also examined 66 patients using 3DFRFSE sequence and assessed the visualization of their pathologies. In a water phantom study, signal intensity (SI) on 3DFRFSE was higher than that on 3DFSE at the same TR ranging from 1,500 to 6,000 ms. In normal volunteers, 3DFRFSE with TR of 2,800 ms showed comparable SI, and signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) superior to those on 3DFSE with TR of 5,000 ms. In clinical setting, 3DFRFSE was useful in demonstrating anatomic details in the labyrinth and pathologic findings of inner ear. The 3DFRFSE can provide high-resolution heavily T2-weighted images (T2WI) with shorter scan time than 3DFSE without significant disadvantage. The 3DFRFSE is a beneficial technique for evaluation of lesions in the inner ear as well as the facial and vestibulocochlear nerves.

Ear, Inner↗
PubMed2002

Salivary cystatins influence ingestion of capsaicin-containing diets in the rat.

Dietary capsaicin consumed by rats over several days induces cystatin-like substances in submandibular saliva. Yet the physiological role of these salivary proteins has not been thoroughly investigated. Salivary cystatins in the rat submandibular glands are known to be induced by chronic treatment with the sympathetic beta-agonist, isoproterenol. In the present study, the possible roles of the salivary proteins on food intake were examined by comparing consumption of a capsaicin-adulterated (0.05%) diet in rats with and without isoproterenol pretreatment (0.1 and 5.0 mg/kg, 5 days). Electrophoretic analysis performed prior to feeding trials revealed that the group pretreated with 5 mg/kg isoproterenol had large amounts of cystatin in the saliva compared with the group pretreated with 0.1 mg/kg isoproterenol and control group. The group treated with 5 mg/kg isoproterenol showed greater consumption of the capsaicin-adulterated diet than the other groups until the 3rd day of trials. Bilateral removal of the submandibular and sublingual glands neutralized the effects of isoproterenol. Induction of salivary cystatins by isoproterenol treatment was not mimicked by systemic and intragastric administration of capsaicin. These results suggest that cystatins are included in the salivary proteins induced by capsaicin and that they contribute to enhanced ingestion of the capsaicin diet. Induction of salivary cystatins may be triggered by irritation of the oral mucosa by capsaicin.

Analysis of Variance↗
PubMed2002

Room temperature phosphorescence of 6-bromo-2-naphthol in micelles of poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) block copolymers and in their mixed aggregates with sodium dodecyl sulfate.

Room temperature phosphorescence (RTP) of 6-bromo-2-naphthol has been investigated in aqueous micellar solutions of poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) block copolymers as well as in their mixed aggregates with sodium dodecyl sulfate. RTP of the phosphorophor was enhanced to some extent in the micelles of the block copolymers. However, marked enhancement of RTP was observed in the mixed aggregates. The enhancement of RTP is attributed to effective incorporation of the phosphorophor into the micelles and the aggregates, resulting in suppression of nonradiative deactivation of the phosphorescent state.

Micelles↗
PubMed2002

New endo-beta-1,4-glucanases from the parabasalian symbionts, Pseudotrichonympha grassii and Holomastigotoides mirabile of Coptotermes termites.

Abstract. An endo-beta-1,4-glucanase (EG) was purified from the hindgut of an Australian mound-building termite, Coptotermes lacteus. The hindgut extract had a peak separate from those for extracts obtained from the salivary glands and the midgut based on sephacryl S-200 gel chromatography, and also demonstrated an origin different from the endogenous EGs of the termite itself. The recovery was further purified by SDS-PAGE, and its N-terminal amino acid sequence analyzed. This showed high homology to EGs from glycoside hydrolase family (GHF) 7. PCR-based cloning methods were applied to the hindgut contents of C. lacteus and individual protozoan symbionts from C formosanus. cDNAs encoding putative EGs homologous to GHF7 members were then identified. The functionality of one of the putative proteins was confirmed by its expression in Escherichia coli.

Amino Acid Sequence↗
PubMed2002

Dual cellulose-digesting system of the wood-feeding termite, Coptotermes formosanus Shiraki.

The distribution of endo-beta-1,4-glucanase (EG) components in the digestive system of the wood-feeding termite, Coptotermes formosanus Shiraki, was investigated by zymogram analysis using polyacrylamide gel electrophoresis, followed by N-terminal protein sequencing. EG components similar to glycoside hydrolase family (GHF) 9 members were restricted to the salivary glands, the foregut, and the midgut, whereas components similar to GHF7 members were confined to the hindgut where numerous cellulolytic flagellates were harbored. RT-PCR experiments revealed that five GHF9 EG mRNAs (1348 bp) homologous to other termite EGs were expressed in the salivary glands and the midgut. The crude extract prepared from the midgut as well as that from the hindgut produced glucose from crystalline cellulose. These data suggest that C. formosanus has two independent cellulose-digesting systems: one in the midgut where cellulose digestion is accomplished by endogenous cellulases and the other in the hindgut which makes use of other cellulases possibly from symbiotic flagellates.

Acrylic Resins↗
PubMed2002

Zolmitriptan is effective and well tolerated in Japanese patients with migraine: a dose-response study.

This phase II study investigated the efficacy, tolerability and dose-response relationship of oral zolmitriptan in the treatment of a single migraine attack in Japanese patients. A bridging analysis then assessed the validity of extrapolating western clinical data to these Japanese patients. In this multicentre, randomized, double-blind, placebo-controlled study, patients received a single dose of placebo or zolmitriptan 1, 2.5 or 5 mg. The primary endpoints were 2-h headache response and the tolerability of zolmitriptan. A statistically significant dose-response relationship was observed for the 2-h headache response (P=0.003). The 2.5 mg group had significantly greater 2-h headache response than the placebo group (P=0.032). The adverse event profile was similar to that reported in western patients, and no adverse events unique to the Japanese population were observed. The bridging analysis report confirmed similar efficacy and tolerability of zolmitriptan in Japanese and western populations. In the Japanese patients, the estimated response rates were 34.3%, 45.2%, 57.7% and 66.2% for placebo, and zolmitriptan 1, 2.5 and 5 mg, respectively, while in the western population the corresponding rates were 39.9%, 49.6%, 61.2% and 71.7%. Zolmitriptan is effective and well tolerated in the acute treatment of migraine in Japanese patients. The optimal dose was 2.5 mg, although the 5 mg dose may provide further benefit for some patients. The bridging analysis supports extrapolation of data from western to Japanese patients.

Adult↗
PubMed2002

Increased cellular infiltrate in inflammatory synovia of osteoarthritic knees.

OBJECTIVE: To determine the nature and origin of synovial inflammation in knees with osteoarthritis (OA). METHOD: Synovial samples were obtained from 21 medial compartmental knee OAs from 19 patients. First, using 11 medial knee synovial samples from 9 patients, a quantitative estimation of synovitis was made with both ordinary and immunohistochemical staining. Second, from the other 10 knees, tissue samples were taken from both the medial and the lateral compartments to quantify cells that infiltrated into the synovium. Medial synovium was immunostained using antibodies to anti-type II collagen, CD68, CD2, CD4, CD8, CD15, CD19, CD25, HLA-DR, CD1a and LN5. The lateral synovium was immunostained with anti-type II collagen, CD68, HLA-DR and CD4 antibody as a control. RESULT: Denatured cartilaginous detritus was found captured by synovial lining cells with a strong immunoreactivity to CD68 antibody, and whose phagocytic potential was activated. The number of anti-type II collagen-positive fragments in the medial compartment of the knee was larger than that found in the lateral compartment. Moreover, the population of CD68-positive cells in synovial tissue and HLA-DR-positive cells in the lining layer was larger in the medial compartment than in the lateral compartment. The number of CD4-positive cells (defined as helper/inducer T lymphocytes) was greater in medial synovium than in lateral synovium. CONCLUSION: Overall, this study strongly supports the concept that the synovitis observed in patients with knee OA might be induced by an immunological mechanism involving, to some extent, a macrophage/helper T cell interaction.

Aged↗
PubMed2002

Distraction lengthening by callotasis in the hand.

The clinical results of 26 digits (18 patients) lengthened by distraction callotasis were evaluated and the factors which influenced healing were analysed. There were 14 men and four women, with a mean age of 39 years. All digits had suffered traumatic amputation. There were eight thumbs and 18 fingers. The level of the site of the osteotomy was at the proximal metaphysis in ten, the middle diaphysis in 13 and the distal metaphysis in three. Although the proposed length was achieved in 23 of the 26 digits, five required additional bone grafts. The rate of healing was 96 days/cm in the digits without complications such as callus fracture or poor callus formation, and 158 days/cm in those with complications. Lengthening at the proximal metaphysis gave a better result than at the diaphysis or distal metaphysis.

Adolescent↗
PubMed2002

Molecular cloning, identification and characterization of four distinct receptor subtypes for insulin and IGF-I in Japanese flounder, Paralichthys olivaceus.

Insulin receptor (IR) and IGF-I receptor (IGF-IR) are structurally and functionally related and belong to the tyrosine kinase receptor family. In teleosti such as salmonids and turbot, occurrence of multiple IR and IGF-IR members has been reported, but the structures of a complete set of both IR and IGF-IR members in a single teleost species have not yet been characterized. In this study, we cloned and analysed four distinct cDNA clones for IR and IGF-IR members from the liver and kidney of the Japanese flounder (Paralichthys olivaceus). Deduced amino acid sequence analyses and phylogenetic analysis have revealed that two of them (fIR-1 and fIR-2) belong to IR members and the other two (fIGF-IR-1 and fIGF-IR-2) are IGF-IRs. fIR-1 and fIR-2 comprised 1369 and 1368 amino acid residues respectively, and fIGF-IR-1 and fIGF-IR-2 comprised 1412 and 1418 residues respectively. All the receptor proteins contained cysteine-rich domains in their alpha-subunits, and conserved each transmembrane and tyrosine kinase domains in their beta-subunits. The amino acid sequences of fIRs and fIGF-IRs showed more than 90% sequence identity with turbot IR and IGF-IR respectively. When compared with their mammalian homologues, fIGF-IR-1 and fIGF-IR-2 proteins contained large insertions at their C-termini, as was observed in the corresponding region of turbot IGF-IR. Occurrence of multiple species of mRNA for each IR and IGF-IR was suggested by Northern blot analyses. A ribonuclease protection assay revealed diverse expressions of four receptor mRNAs in a wide range of tissues including heart, liver, ovary, testis, brain, gill arch, kidney, skeletal muscle, intestine, stomach, spleen and eye of the flounder.

5' Untranslated Regions↗
PubMed2001

Traumatic injury-induced BMP7 expression in the adult rat spinal cord.

It has been reported that bone morphogenetic proteins (BMPs) are involved in the generation of the central nervous system during development. However, the roles of BMPs in mature spinal cord have not been clarified. We examined the expression of BMP7 mRNA before and after traumatic injury of the adult rat spinal cord. BMP7 mRNA was already detectable at a relatively low level in uninjured spinal cord, but was dramatically increased after injury. Semiquantitative RT-PCR study further confirmed upregulation of BMP7 mRNA in injured spinal cord. In situ hybridization indicated that expression of BMP7 mRNA was present only in glial cells in uninjured spinal cord. After injury, the number of BMP7-expressing glial cells was increased, BMP7 expression also became apparent in motor neurons. It has been suggested that BMPs promote survival of subventricular zone cells in adult rats. Thus, our results suggest that increase in the expression of BMP7 promotes survival of neurons and glial cells after acute traumatic injury. In contrast, there is increasing evidence that BMPs inhibit neurogenesis and alternatively promote gliogenesis of neural progenitors, which are also present in adult spinal cord, suggesting that injury-upregulated BMP7 may regulate differentiation of glial cells from neural progenitors and may induce gliosis after central nervous system injury.

Age Factors↗
PubMed2001

Structure of the beta-chain (B29) gene of the chicken B-cell receptor and conserved collinearity with genes for potential skeletal muscle sodium channel and growth hormone.

Abstract. Reported here is the cloning of the gene for the beta-chain (Ig beta) of the chicken B-cell receptor for antigen. A cDNA library made from B lymphocyte-derived DT40 cells was first screened with a beta-chain cDNA clone from rat. A full-length clone (1556 bp) was obtained encoding a 226-amino-acid beta-chain having 40% sequence identity with the highly conserved beta-chains present in mammals and producing mRNA of 1.7 kb. Clones containing the full B29 gene encoding chicken Ig beta were isolated and fully sequenced. As in mammals, the chicken B29 gene consists of six exons. The chicken B29 gene is somewhat larger (4336 bp) than that of rat (3.1 kb) and human (3.6 kb). Genome collinearity was found in the region of the chicken B29 gene. Genes for potential skeletal muscle sodium channel (2.2 kb upstream) and for growth hormone (2.3 kb downstream) were found in the same order and transcriptional orientation as in mammals, though the genes are more closely positioned in the chicken.

Animals↗