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Biomedical subjects

S Kawai

Publications and source records attributed to S Kawai.

At least 55 records · Page 3Linked to original sources
PubMed2001

Vascular complication in free tissue transfer to the leg.

The lower-extremity free flap has a high incidence of vascular complication. A retrospective study of 70 free flap transplants in 70 patients (1987-2000) was conducted to investigate factors leading to vascular complication and free flap failure. The overall success rate was 92% (64 of 70 flaps); the incidence of vascular complications was 22% (16 of 70 flaps). Among 16 complicated flaps, 7 were transferred in severely crushed legs, and 4 were in multioperated legs attributable to chronic osteomyelitis. Venous thrombosis occurred in 12 flaps (86%). The success rate of the patients operated on at Yamaguchi University Hospital was higher (96%) than at other hospitals tested. The key factors contributing to improved outcome in free tissue transplantation in the leg were careful preoperative planning for highly traumatized legs and proper selection of the recipient vein and of the hospital at which immediate reexploration can be performed for vascular complicated flaps.

Adolescent↗
PubMed2001

Activated Ras modifies the proliferative response of rheumatoid synovial cells to TNF-alpha and TGF-alpha.

OBJECTIVE: To study the role of the Ras/mitogen-activated protein kinase (MAPK) pathway in the proliferative response of rheumatoid synovial fibroblast (RSF) to tumor necrosis factor (TNF)-alpha and transforming growth factor (TGF)-alpha. METHODS: V-Ki-ras gene was introduced into RSF using a retrovirus and the proliferative response of these cells to TNF-alpha or TGF-alpha was estimated by measuring the uptake of 3H-thymidine. The effect of a mitogen-activated protein kinase kinase (MEK) inhibitor, PD98059, was also investigated. RESULTS: Consistent with previous reports, TNF-alpha and TGF-alpha stimulated the proliferation of RSF. When the v-Ki-ras gene was expressed, the basal growth rate of these cells was increased, but their growth was suppressed by TNF-alpha or TGF-alpha. The latter effect was abolished when the cells were exposed to a relatively low concentration of PD98059. CONCLUSION: Ras modulates the proliferative response of RSF to TNF-alpha and TGF-alpha.

Apoptosis↗
PubMed2001

Invasive forms of Toxoplasma gondii, Leishmania amazonensis and Trypanosoma cruzi have a positive charge at their contact site with host cells.

We examined the surface charges of invasive forms of Toxoplasma gondii, Leishmania amazonensis, and Trypanosoma cruzi by atomic force microscopy and surface potential spectroscopy. We found that the specific part of the protozoan which makes initial contact with the host cell is positively charged. This indicates that the positive charge at the site of contact facilitates binding of the invasive protozoan to negatively charged host cells.

Animals↗
PubMed2001

Endoscopic carpal tunnel release and nerve conduction studies.

We investigated the outcome of endoscopic carpal tunnel release (ECTR) for patients with carpal tunnel syndrome (CTS) in comparison with the results of preoperative nerve conduction studies. The compound muscle action potential (CMAP) of the abductor pollicis brevis muscle (APB) and the second lumbrical muscle (L2) was recorded following median nerve stimulation at the wrist. A total of 38 hands in 35 patients were classified into four categories. Hands with a similarly prolonged distal motor latency for the APB and L2 were classified as type I (n=25), while those with a more prolonged distal motor latency for the APB than for the L2 (>0.7 ms) were classified as type 2 (n=10). Hands with a CMAP for the APB, but not L2, were classified as type 3 (n=1), and hands with no CMAP for either the APB or L2 were classified as type 4 (n=2). After ECTR, all of the type 1 and 2 hands were improved. Patients with type 3 and type 4 hands did not show satisfactory improvement, which may have been due to anatomical variation of the recurrent motor branch of the median nerve.

Action Potentials↗
PubMed2001

Two novel Pao-like retrotransposons (Kamikaze and Yamato) from the silkworm species Bombyx mori and B. mandarina: common structural features of Pao-like elements.

To characterize the structural features common to Pao-like retrotransposons, we analyzed two lambda phage clones which contain the Pao-like elements from the silkworm species Bombyx mori and B. mandarinia, and copies of Pao itself and ninja of Drosophila simulans, amplified by PCR. We previously identified two randomly amplified polymorphic DNAs (RAPDs), W-Kamikaze and W-Yamato, from B. mori and B. mandarina, which are part of two novel Pao-like retrotransposons, Kamikaze and Yamato, respectively. Complete characterization of these and other elements of this group reported here shows that Pao-like elements have common features that distinguish them from the other groups of LTR-retrotransposons. While the elements of the Ty1-copia group encode only one cysteine and histidine (Cys) motif in their gag-like region, the Pao-like elements specify three Cys motifs. The highly conserved D(35)E motif in the integrase domain of the retrotransposon polyprotein seems to be conserved in Pao-like elements, but the number of amino acid residues between D and E varies and is greater than 35. A comparison of the deduced amino acid sequences of the reverse transcriptase domain revealed the Pao-like elements are members of neither the Ty1-copia nor the gypsy-Ty3 groups. Therefore, we confirmed that the long-terminal-repeat (LTR) retrotransposons should be divided into three major groups (or families), namely the Ty1-copia, gypsy-Ty3, and Pao-like groups.

Amino Acid Sequence↗
PubMed2001

Aortic and mitral valve replacement in a patient with acute febrile neutrophilic dermatosis (Sweet's syndrome): report of a case.

A 29-year-old woman was admitted to our hospital with severe orthopnea, fever, and acute dermatosis. She had a 5-year history of episodic acute neutrophilic dermatosis and peripheral leukocytosis following a high fever, which were symptoms consistent with a diagnosis of Sweet's syndrome. Echocardiography revealed remarkable dysfunction of the left ventricle due to severe aortic regurgitation, which had not been present at a previous admission when mild mitral regurgitation was detected. The aortic and mitral valves were replaced with prosthetic valves on an emergency basis. The leaflets of the aortic valve were very thin and appeared fragile. The anterior leaflet of the mitral valve showed severe prolapse due to the torn chordae and hypoplasia of the posterior strut chordae. Her postoperative course was uneventful. Microscopic examination revealed fibrosal degeneration and the infiltration of lymphocytes and macrophages into both heart valves. This may be the first case report of valvulitis and Sweet's syndrome occurring simultaneously.

Adult↗
PubMed2001

Possible involvement of vitamin D receptor gene polymorphism in male patients with ossification of spinal ligaments.

Ossification of spinal ligaments (OSL) is a common form of myelopathy characterized by heterotopic bone formation in the spinal ligaments, predominantly in men. Although the etiology of OSL is not fully understood, previous studies have strongly suggested the involvement of genetic factors in this disease. To investigate the possible involvement of vitamin D receptor (VDR) gene polymorphism in Japanese male patients with OSL, we analyzed: (a) the VDR genotype defined by BsmI polymorphism in patients with obvious OSL and controls; and (b) the effect of 1,25-dihydroxyvitamin D3 on alkaline phosphatase (ALP) activity of spinal ligament cells derived from patients without OSL. With regard to the VDR genotype, of the patients with OSL (n = 27), none had the BB genotype (0%), one had the Bb genotype (4%), and 26 had the bb genotype (96%). In the control group (n = 97) three had the BB genotype (3%), 18 had the Bb genotype (19%), and 76 had the bb genotype (78%). As a result, the B allele frequency in patients with OSL (2%) was significantly lower than in controls (12%). 1,25-Dihydroxyvitamin D3, at concentrations of 10-9 and 10-8 M, significantly increased ALP activity of the ligament cells (n = 8), suggesting that 1,25-dihydroxyvitamin D3 is able to promote osteogenic differentiation of normal ligament cells. Among the Japanese, sensitivity to vitamin D has been reported to vary between the alleles of the VDR; i.e., bone mineral density (BMD) in patients without the B allele is increased by vitamin D treatment, whereas patients with the B allele do not show such an increase in BMD. The present investigation is a small preliminary study, but the findings suggest, for the first time, that the B allele of the VDR acts as an inhibitor in the pathogenesis of human male OSL.

24,25-Dihydroxyvitamin D 3↗
PubMed2001

Carboxylation of osteocalcin may be related to bone quality: a possible mechanism of bone fracture prevention by vitamin K.

Vitamin K is essential for the carboxylation of glutamic acid residues, such as osteocalcin. Recent studies have reported that vitamin K reduces vertebral and hip fractures without increasing bone mass in patients with osteoporosis, suggesting that vitamin K could affect bone quality. However, the mechanism is unknown. To investigate the involvement of the carboxylation of osteocalcin in bone quality, the present preliminary study examined serum bone markers and ultrasound velocity, a possible indicator of bone quality, in 14 healthy prepubertal children (eight boys and six girls) aged between 7 and 12 years. Venous blood was collected between 0800 and 0900 h after an overnight fast, and serum levels of intact, carboxylated and undercarboxylated osteocalcin, bone-specific alkaline phosphatase and type I procollagen carboxyl extension peptide were measured. Speed of sound in the right tibia was measured using a SoundScan 2000 Compact (Myriad Ultrasound System, Rehovot, Israel). As a result, there was no significant correlation between the serum bone markers and the Z score for the speed of sound. In contrast, the ratio of serum carboxylated osteocalcin to serum intact osteocalcin was positively correlated with the Z score for the speed of sound (r = 0.621, P = 0.016). These findings suggest, for the first time, that carboxylation of osteocalcin is related to bone quality. Further studies are needed to clarify the role of carboxylation of osteocalcin in bone, and this will provide a new insight into the mechanism of vitamin K treatment in osteoporosis.

Alkaline Phosphatase↗
PubMed2001

Molecular characterization of a 2-Cys peroxiredoxin from the human malaria parasite Plasmodium falciparum.

We have identified the 2-Cys peroxiredoxin (PfPrx-1) from the human malaria parasite Plasmodium falciparum. The PfPrx-1 showed the highest identity at amino acid level to the type II Prx among the currently known six subfamilies of mammalian Prx. The sequence identity between the PfPrx-1 and the previously reported 1-Cys Prx of P. falciparum (PfPrx-2), which corresponded to mammalian type VI Prx, was 25%. This suggests that the parasite possesses two Prx subfamilies. The PfPrx-1 showed significant sequence similarities with those of 2-Cys peroxiredoxins of plants in the BLASTX search. This may reflect the consequences of a genetic transfer from an algal endosymbiont to the parasite nucleus during evolution. The recombinant PfPrx-1 protein (rPfPrx-1) was expressed as a histidine fusion protein in Escherichia coli and purified with Ni chromatography. The rPfPrx-1 existed as dimers under non-reducing conditions and dissociated into monomers in the presence of dithiothreitol. The PfPrx-1 protein also exists as a dimer in the parasites themselves. The reduction of the oxidized enzyme by the donation of electrons from E. coli thioredoxin (Trx)/Trx reductase system was demonstrated in its reaction with H(2)O(2), using the rPfPrx-1 protein. These results suggested that the PfPrx-1 can act as a terminal peroxidase of the parasite Trx system. An elevated expression of the PfPrx-1 protein seen in the trophozoite, the stage with active metabolism, suggests an association of the parasite Trx system with its intracellular redox control.

Amino Acid Sequence↗
PubMed2001

A new rabbit model for the study on cervical compressive myelopathy.

Development process and pathology of myelopathy due to chronic spinal cord compression have not been fully elucidated. This study was conducted in order to establish an experimental model which can efficiently produce myelopathy and be useful in the studies on myelopathy due to chronic spinal cord compression. Under electrophysiological monitoring of the spinal cord, anterior compression was produced on C5 using a plastic screw. Two weeks later, a plastic plate was inserted under the C5 arch. For the subsequent 10 months on average, walking pattern and MR images were periodically monitored. Before the sacrifice, electrophysiological test was performed and then histopathological examination was done. Palsy appeared at 5 months on average after the addition of posterior compression. Mean compression ratio of the spinal cord calculated on MR images was 34%. All animals with compression showed a high intramedullary signal intensity, and the mean contrast-to-noise ratio (CNR) in the compressed area was 49%. Electrophysiological test showed a significant decrease in the amplitude of spinal cord evoked potentials (SCEPs) at the given compression level. Histology showed flattening of the anterior horn, disappearance and necrosis of anterior horn cells in the gray matter; and demyelination and axonal degeneration in the white matter. The antero-posterior compression produces the condition of spinal canal stenosis. Repeated antero-posterior compression to the spinal cord is important in establishing myelopathy. The present animal model was evaluated to be useful in the studies on myelopathy.

Animals↗
PubMed2001

Mechanism of prolonged central motor conduction time in compressive cervical myelopathy.

OBJECTIVE: To investigate the mechanism of prolonged central motor conduction time (CMCT) in compressive cervical myelopathy, we compared the calculated CMCT following transcranial magnetic stimulation (TCM) and evoked spinal cord potentials (ESCPs) following transcranial electric stimulation (TCE). METHOD: Motor evoked potentials following TCM were recorded from abductor digiti minimi and abductor hallucis brevis muscles in 16 patients with compressive cervical myelopathy. CMCT was calculated by subtracting peripheral conduction time using peripheral nerve stimulation from MEP latency. ESCPs following TCE were recorded intraoperatively from posterior epidural space. RESULTS: CMCT was prolonged and significant attenuation of the ESCP amplitude following TCE was observed in all patients with cervical myelopathy. In 8 of 16 patients CMCT was significantly prolonged but ESCPs were recorded at the C6-7 level with normal negative peak latency. CONCLUSIONS: Prolonged CMCT may occur with only a minor amount of conduction slowing in the corticospinal tract in compressive cervical myelopathy. Impaired temporal summation of multiple descending potentials following TCM produced delays of motor neuron firing that contribute to the mechanism of prolonged CMCT.

Aged↗
PubMed2001

Activation of mitogen-activated protein kinase cascades is involved in regulation of bone morphogenetic protein-2-induced osteoblast differentiation in pluripotent C2C12 cells.

Bone morphogenetic protein (BMP)-2, a member of the transforming growth factor-beta (TGF-beta) superfamily, is able to induce osteoblastic differentiation of C2C12 cells. Both Smad and mitogen-activated protein kinase (MAPK) pathways are essential components of the TGF-beta superfamily signaling machinery. Although Smads have been demonstrated to participate in the BMP-2-induced osteoblastic differentiation of C2C12 cells, the role of MAPK has not been addressed. This report shows that BMP-2 activates ERK and p38, but not JNK, in C2C12 cells. Pretreatment of cells with the p38 inhibitor, SB203580, dramatically reduced BMP-2-induced expression of the osteoblast markers alkaline phosphatase (ALP) and osteocalcin (OC). Nevertheless, overexpression of MKK3, a protein kinase that phosphorylates and activates p38, failed to induce ALP or OC expression in the absence of BMP-2, indicating that p38 activation is necessary but not sufficient for the acquisition of the osteoblast phenotype by these cells. Although ALP induction was increased slightly in the presence of PD-98059, a selective inhibitor of the ERK cascade, this compound significantly inhibited both steady-state and BMP-2-induced OC RNA levels. Our results indicate that p38 and ERK cascades play a crucial role in the osteoblast differentiation of C2C12 cells mediated by BMP-2.

Alkaline Phosphatase↗
PubMed2001

Characterization of human bone morphogenetic protein (BMP)-4 and -7 gene promoters: activation of BMP promoters by Gli, a sonic hedgehog mediator.

Among the bone morphogenetic protein (BMP) family, which plays a crucial role not only in bone formation but also in development, BMP-2, -4, and -7 participate predominantly in various aspects. To undertake complex tasks, their expression is strictly controlled. In this study we isolated and analyzed the 5'-flanking regions of the human BMP-4 and -7 genes to elucidate the mechanism of their temporally and spatially specific expression. As for BMP-4 expression, a reverse transcription-polymerase chain reaction (RT-PCR) assay with specially designed sets of primers demonstrated that osteoblastic SaOS-2 and Hos cells expressed two types of transcripts comprising one of the 5'-untranslated first exons, whereas MG63 cells displayed only the transcript with the BMP-4 proximal first exon. Likewise, RT-PCR revealed that Hos and MG63 cells expressed BMP-7. Subsequent 5'-RACE confirmed an alternative usage of the BMP-4 first exons with clustered multiple transcription start sites in the distal exon and the sole start site in the proximal exon. The transcription start site of the BMP-7 gene was found to be far upstream (764 bp) of the initiation ATG codon. We constructed a series of deletion mutants of fusions between these BMP promoters and the luciferase gene and examined their activity by transient transfection into osteoblastic Hos and renal COS-7 cells. The degree of distal and proximal BMP-4 promoter activity was in accordance with the expression level of the corresponding transcripts. Both distal and proximal BMP-4 promoters possessed suppressor elements that are operative only in Hos cells. The positive and negative elements identified in the BMP-7 promoter were more remarkably effective in Hos cells. The activities of the respective BMP-4 promoters and BMP-7 promoter were all stimulated upon the cotransfection of a potential sonic hedgehog (SHH) mediator, Gli1 or Gli3 into COS-7 cells, providing direct evidence that the Gli proteins are capable of inducing the BMP expression. Our systems are helpful for assessment of the complicated interactions of molecules involved in the skeletogenesis and developmental processes.

Animals↗
PubMed2001

Opposite effects of bone morphogenetic protein-2 and transforming growth factor-beta1 on osteoblast differentiation.

Several members of the transforming growth factor-beta (TGF-beta) superfamily have been demonstrated to play regulatory roles in osteoblast differentiation and maturation, but the mechanisms by which they act on different cells at different developmental stages remain largely unknown. We studied the effects of TGF-beta1 and bone morphogenetic protein-2 (BMP-2) on the differentiation/maturation of osteoblasts using the murine cell lines MC3T3-E1 and C3H10T1/2. BMP-2 induced or enhanced the expression of the osteoblast differentiation markers alkaline phosphatase (ALP) and osteocalcin (OC) in both cells. In contrast, TGF-beta1 was not only unable to induce these markers, but it dramatically inhibited BMP-2-mediated OC gene expression and ALP activity. In addition, TGF-beta1 inhibited the ability of BMP-2 to induce MC3T3-E1 mineralization. TGF-beta1 did not sensibly modify the increase of Osf2/Cbfa1 gene expression mediated by BMP-2, thus demonstrating that the inhibitory effect of TGF-beta1 on osteoblast differentiation/maturation mediated by BMP-2 was independent of Osf2/Cbfa1 gene expression. Finally, it is shown that TGF-beta1 does not affect BMP-2-induced Smad1 transcriptional activity in the mesenchymal pluripotent cells studied herein. Our data indicate that in vitro BMP-2 and TGF-beta1 exert opposite effects on osteoblast differentiation and maturation.

Alkaline Phosphatase↗
PubMed2001

Allele frequency of HLA-B39 in the Japanese population and identification of a novel B39 allele, B*3923.

A new HLA-B39 allele, B*3923, was found in the Japanese population. Compared with B*39022, the new allele had a single point mutation at position 503 in exon 3 with an amino acid substitution, Gln144Arg. To determine B39 allele frequency in Japanese, 275 B39-positive samples from 3277 Japanese individuals were examined by polymerase chain reaction using microtitre plate hydribization (PCR-MPH) and single-strand conformation polymorphism (PCR-SSCP). The frequency of B*3923 was estimated to be 0.4% in B39 alleles of the Japanese population. B*3923 was found in samples from four Japanese individuals, all of whom have DRB1*1406.

Asian People↗
PubMed2001

Selective recruitment of CCR6-expressing cells by increased production of MIP-3 alpha in rheumatoid arthritis.

Infiltration of various types of leucocytes has been shown to play a crucial role in the pathogenesis of rheumatoid arthritis (RA). Macrophage inflammatory protein-3 alpha (MIP-3 alpha) is a recently identified chemokine which is a selective chemoattractant for leucocytes such as memory T cells, naïve B cells and immature dendritic cells. In this study, we investigated the expression of MIP-3 alpha and its specific receptor CCR6 in the inflamed joints of patients with RA. Increased amounts of MIP-3 alpha were found by ELISA in synovial fluids (SF) of patients with RA. MIP-3 alpha was apparently detected in all synovial tissue specimens of RA patients (n = 6), but it could not be detected in that of osteoarthritis (OA) patients (n = 4). Expression of MIP-3 alpha was detected especially in the sublining layer, and infiltrating mononuclear cells in RA synovial tissue. Gene expression of MIP-3 alpha was also found in six out of 11 RA-synovial fluid cells by RT-PCR. Cultured synovial fibroblasts derived from either RA or OA patients were capable of producing MIP-3 alpha in response to IL-1 beta and TNFalpha in vitro. Furthermore, expression of CCR6 was found in infiltrating mononuclear cells in the cellular clusters and around the vessels of RA synovial tissue. These findings indicate that increased production of MIP-3 alpha may contribute to the selective recruitment of CCR6-expressing cells in RA.

Arthritis, Rheumatoid↗
PubMed2001

Deduced amino-acid sequence of a calcium-free alpha-amylase from a strain of Bacillus: implications from molecular modeling of high oxidation stability and chelator resistance of the enzyme.

Alkaline alpha-amylase (AmyK38) from the alkaliphilic Bacillus sp. strain KSM-K38 is a unique enzyme in that it is highly chelator-resistant and oxidatively stable [Hagihara, H., Igarashi, K., Hayashi, Y., Endo, K., Ikawa-Kitayama, K., Ozaki, K., Kawai, S. & Ito, S. (2001) Appl. Environ. Microbiol. 67, 1744-1750]. This enzyme was found to contain no Ca and require Na (or monovalent cations) for manifestation of activity. The nucleotide sequence of the gene for the novel enzyme was determined, and it harbored an ORF of 1503 bp encoding the enzyme of 501 amino acids, including a 21-amino-acid signal peptide. The deduced amino-acid sequence of the mature enzyme (55 097 Da) showed moderate homology to those of alpha-amylases from Bacillus licheniformis, Bacillus stearothermophilus and Bacillus amyloliquefaciens, with approximately 63% identity. A methionine residue, which is conserved and susceptible to chemical oxidation, was replaced with leucine in AmyK38. Moreover, many conserved residues that are crucial ligands for Ca were replaced with other amino acids, thereby leading to loss of the Ca coordination geometries. By building a molecular model, we showed the calcium-independent, oxidatively stable active-site topology and structural integrity of AmyK38.

Alkalies↗