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Biomedical subjects

J Machida

Publications and source records attributed to J Machida.

At least 19 recordsLinked to original sources

Complete sequencing shows a role for MSX1 in non-syndromic cleft lip and palate.

MSX1 has been proposed as a gene in which mutations may contribute to non-syndromic forms of cleft lip and/or cleft palate. Support for this comes from human linkage and linkage disequilibrium studies, chromosomal deletions resulting in haploinsufficiency, a large family with a stop codon mutation that includes clefting as a phenotype, and the Msx1 phenotype in a knockout mouse. This report describes a population based scan for mutations encompassing the sense and antisense transcribed sequence of MSX1 (two exons, one intron). We compare the completed genomic sequence of MSX1 to the mouse Msx1 sequence to identify non-coding homology regions, and sequence highly conserved elements. The samples studied were drawn from a panethnic collection including people of European, Asian, and native South American ancestry. The gene was sequenced in 917 people and potentially aetiological mutations were identified in 16. These included missense mutations in conserved amino acids and point mutations in conserved regions not identified in any of 500 controls sequenced. Five different missense mutations in seven unrelated subjects with clefting are described. Evolutionary sequence comparisons of all known Msx1 orthologues placed the amino acid substitutions in context. Four rare mutations were found in non-coding regions that are highly conserved and disrupt probable regulatory regions. In addition, a panel of 18 population specific polymorphic variants were identified that will be useful in future haplotype analyses of MSX1. MSX1 mutations are found in 2% of cases of clefting and should be considered for genetic counselling implications, particularly in those families in which autosomal dominant inheritance patterns or dental anomalies appear to be cosegregating with the clefting phenotype.

Amino Acid Sequence↗

Development of Dunn osteosarcoma inoculated into subcutaneous air pouch in mice.

Dunn osteosarcoma cell growth was observed in C3H/Hej mouse subcutaneous air pouch. Inoculation with Dunn osteosarcoma cell suspension was performed by injection into air pouches of 70 mice on Day 7 after the initial injection of air. The developmental pattern of the tumor cell colonies was classified histologically into five stages. In Stage 0 (stage of no tumor cells), there were no tumor cells. In Stage 1 (focal stage), the colonies were limited to the lining-cell layers. In Stage 2, (segmental stage) the colonies protruded into the cavity of the pouch. In Stage 3 (annular stage), the total surface of the inner wall of the pouch was occupied by tumor cells. In Stage 4 (occlusive stage), the cavity of the pouch was fully occupied by tumor cells. The tumor was observed to develop to stage 1 in all mice 7 days after inoculation. The colonies were all found to be settled initially in the deeper layer of lining cells. Thus, the mouse air-pouch model is useful for direct observation of osteosarcoma cell growth.

Animals↗

[Clinical features of patients with osteoarthritic knees followed by development of rheumatoid arthritis].

We investigated clinical features of patients with osteoarthritic knees followed by development of rheumatoid arthritis (RA) after several year's interval. The subjects were 16 knees of 8 patients with osteoarthritis (OA) including one man and 7 women. The mean age at development of OA knee was 62.8 years (range; 45-73). The mean age at later development of RA was 66.0 years (range; 52-79). The mean follow-up period was 96.4 months (range; 28-191). We evaluated clinical features using the 1987 revised Criteria of the American College of Rheumatology (ACR), laboratory dates including RF, CRP, ESR, the number of joints with RA, and femorotibial angle (FTA). The mean number of features of patients which was fulfilled with the ACR criteria was 3.3 +/- 1.6 at the onset of RA. Only four patients were seropositive through the total follow-up period. The serum level of RF, CRP, and ESR were reduced at the follow-up period. The mean number of the joints involved in RA was 11.0 +/- 5.1 (range; 4-22) and wrist and shoulder joints were involved more frequently than other joints except knees. High tibial osteotomy (HTO) was performed on 5 knees of 3 patients and the mean degree of FTA was 168.8 +/- 1.9 degrees just after surgery. However, 36 months after development of RA, joint destruction and valgus deformity occurred on 3 knees and the mean degree of FTA of 5 knees was ended up to 159.6 +/- 11.3 degrees. Our experiences suggested that RF, CRP, ESR and lesions of other joints should be carefully evaluated in the OA patients with seropositivity or knee hydrarthrosis and that histological analysis for synovium should be assessed by the biopsy at time of HTO or arthroscopic surgery to improve accuracy of diagnosis.

Aged↗

Transforming growth factor-alpha (TGFA): genomic structure, boundary sequences, and mutation analysis in nonsyndromic cleft lip/palate and cleft palate only.

Transforming growth factor-alpha (TGFA) has been proposed as a candidate gene in the etiology of nonsyndromic cleft lip with or without cleft palate (NS-CL/P) and of nonsyndromic cleft palate only (NS-CPO). Biologic support for a role of TGFA arises from its presence at high levels in the epithelial tissue of the medial edge of the palatal shelves at the time of shelf fusion in mice. Genetic support for the role of TGFA in clefting comes from the reported association of TGFA alleles with human NS-CPO and NS-CL/P. In this study we report the sequence and structure of human genomic TGFA and the search for causal TGFA mutations in 250 individuals with NS-CL/P or NS-CPO by conformational analysis of the coding sequence, splice junctions, and a portion of the 3' untranslated region strongly homologous between human and mouse. We confirm that human TGFA is composed of six exons and here report several new sequence substitutions and their frequencies. Five variants in conserved segments may represent rare causes for clefting in humans and provide support for the role of TGFA in facial morphogenesis.

Animals↗

Highly activated matrix metalloproteinase-2 secreted from clones of metastatic lung nodules of nude mice injected with human fibrosarcoma HT1080.

The promoting effects of matrix metalloproteinase-2 (MMP-2) on lung metastasis of human fibrosarcoma cells (HT1080) were studied using nude mice. The fourth generation of HT1080 was established by consecutive clonal selection of metastatic lung nodules formed by intravenous transplantation. MMP-2 and MMP-9 in the culture supernatants of the first and fourth generation cells were analyzed by gelatin zymography and Western blotting, and quantified by scanning densitometry. In gelatin zymograms, mean ratios of values for the 59-kDa band (the active form of MMP-2) to those for the 72-kDa band (the inactive form of MMP-2) for optical density; area, and volume measured by densitometry were 1.44 +/- 0.12, 0.93 +/- 0.05, and 1.27 +/- 0.20, respectively, in the culture supernatant of fourth generation cells isolated from metastatic lung nodules. These values were significantly (P < 0.01) higher than those of first generation cells (0.70 +/- 0.04, 0.48 +/- 0.01, and 0.57 +/- 0.42). Three weeks after intravenous transplantation of HT1080 cells into nude mice, the incidence of lung metastasis and mean number and diameter of metastatic nodules formed by injection of first generation cells were 20% (2 of 10 mice), 2.9 +/- 0.2 and 2.0 +/- 0.2 mm, respectively; while they were 100%, 99.8 +/- 7.2 and 4.3 +/- 0.3 mm following injection of fourth generation cells. These findings suggest that the active MMP-2 produced by human fibrosarcoma cells is important for the cells to form lung metastatic lesions in nude mice.

Animals↗

Effects of lipo prostaglandin E1 on distribution of cis-diamminedichloroplatinum in lung metastasis derived from Dunn osteosarcoma cell-inoculated mouse foot-pad.

This study was designed to determine the effect of lipid microspheres containing prostaglandin E1 (lipo PGE1) on cis-diamminedichloroplatinum (CDDP) accumulation in primary and lung metastatic lesions. Sixty mice were divided into four groups, depending on whether or not an intra-foot-pad injection of Dunn osteosarcoma cells had been administered and on whether or not an intraperitoneal injection of lipo PGE1 had been administered. CDDP was injected intraperitoneally into all the mice 6 weeks after the inoculation. Tumor colonies of spontaneous metastases in the left lung were found in 21 out of 30 tumor-inoculated mice at autopsy. Tissue platinum concentrations in the lungs with metastatic colonies and in the foot-pad tumors were significantly higher in the lipo PGE1-administered mice than in those without treatment. Terminal deoxytransferase-mediated dUTP nick end labeling (TUNEL) assay showed marked localization of dying cells in the lung metastatic lesions of the lipo PGE1-administered mice. The results of this study showed that pretreatment with lipo PGE1 may augment the antitumor effects of CDDP at the tumor site.

Alprostadil↗

Associations of medical status and physical fitness with periodontal disease.

To determine the possible associations of medical status and physical fitness with periodontal disease, a cross-sectional study was conducted. The subjects were 517 males and 113 females aged 23 to 83 years who participated in a multiphasic health test at the Aichi Prefectural Center of Health Care, Japan, from 1992 to 1997. Their periodontal status was assessed by means of the CPITN scoring system. To assess the strength of associations between the examined factors and the score, odds ratios were computed using ordinal logistic models. Conventional risk factors such as old age, smoking habits, and higher fasting plasma glucose and simplified debris index increased the risk of periodontal disease. Hypertension, hematuria, leucocytosis or thrombocytosis, positive C-reactive protein and higher serum alkaline phosphatase were positively associated with the score, whereas higher serum high-density lipoprotein cholesterol was related to a lower risk. Poor physical fitness affecting aerobic capacity, foot balance and reaction was associated with a higher CPITN score. These associations were independent of the conventional risk factors. Although these new potential risk factors should be further investigated for their causal relationship, our findings suggested a close relationship of oral health to medical status and physical fitness.

Adult↗

Characterization of a novel gene disrupted by a balanced chromosomal translocation t(2;19)(q11.2;q13.3) in a family with cleft lip and palate.

Cleft lip with or without cleft palate is a common birth defect that is genetically complex. The nonsyndromic forms have been studied genetically using linkage and candidate-gene association studies with only partial success in defining the loci responsible for orofacial clefting. Loci for nonsyndromic cases have been suggested on 2p13, 4q31, 6p24, 17q21-q24, and 19q13.2. Recently, we identified a family in which cleft lip and palate segregated in two of three generations with a balanced chromosomal translocation t(2;19)(q11. 2;q13.3). We used a positional-cloning strategy to identify a novel gene disrupted by the translocation on chromosome 19. Eight rare (q < 0.01) and nine common (q > 0.01) variants of this gene were detected in the DNA of 74 unrelated cases of cleft lip and/or cleft palate; no variants associated significantly with clefting, suggesting that this gene is not a major contributor to abnormal craniofacial development. This gene, CLPTM1, was ubiquitously expressed on Northern blots containing RNA from adult tissues and in whole-mount in situ hybridization of day 10 to 12 mouse embryos. CLPTM1 encodes a transmembrane protein and has strong homology to two Caenorhabditis elegans genes, suggesting that CLPTM1 may belong to a new gene family.

Alleles↗

Association of MSX1 and TGFB3 with nonsyndromic clefting in humans.

Nonsyndromic cleft lip with or without cleft palate (CL/P) and nonsyndromic cleft palate only (CPO) are common congenital anomalies with significant medical, psychological, social, and economic ramifications. Both CL/P and CPO are examples of complex genetic traits. There exists sufficient evidence to hypothesize that disease loci for CL/P and CPO can be identified by a candidate-gene linkage-disequilibrium (LD) strategy. Candidate genes for clefting, including TGFA, BCL3, DLX2, MSX1, and TGFB3, were screened for LD with either CL/P or CPO in a predominantly Caucasian population, with both case-control- and nuclear-family-based approaches. Previously reported LD for TGFA with both CL/P and CPO could not be confirmed, except in CL/P patients with a positive family history. Also, in contrast to previous studies, no LD was found between BCL3 and either CL/P or CPO. Significant LD was found between CL/P and both MSX1 and TGFB3 and between CPO and MSX1, suggesting that these genes are involved in the pathogenesis of clefting. In addition, a mutation search in the genes DLX2, MSX1, and TGFB3 was performed in 69 CPO patients and in a subset of the CL/P patients. No common mutations were found in the coding regions of these genes; however, several rare variants of MSX1 and TGFB3 were found that may alter the latters' normal function. These results form the basis for future research, including (a) mutation searches in the MSX1 and TGFB3 genes in Caucasian CL/P patients and (b) extension of the search for MSX1 mutations in CPO patients to the noncoding regions.

Amino Acid Substitution↗

Natural course of untreated symptomatic temporomandibular joint disc displacement without reduction.

In some patients with disc displacement without reduction, the symptoms of pain and decreased range of motion have been observed to resolve spontaneously over time without treatment. The natural history of this condition, however, is not well-understood. Thus, to study the natural course of disc displacement without reduction, we followed 40 patients without treatment for a period of 2.5 years. The diagnosis was established by history and physical examination and confirmed with magnetic resonance (MR) imaging. After 2.5 years, 43% of the patients were asymptomatic, 33% had decreased symptoms, and 25% of the patients showed no improvement or had required treatment. MR evidence of osteoarthritis and advanced stages of internal derangement at the initial evaluation was associated with a poor prognosis. The result of this prospective cohort study indicated that approximately 40% of patients with symptomatic disc displacement without reduction will be free of symptoms within 2.5 years, one-third will improve, whereas one-quarter will continue to be symptomatic. This knowledge should be valuable for the treatment planning and evaluation of prognosis of patients with non-reducing symptomatic disc displacement.

Adolescent↗

Effect of cyclosporin A on isolated rabbit bladder and urethral smooth muscle.

BACKGROUND: The effects of Cyclosporin A (CsA) on vascular smooth muscle inducing hypertension have been established. However, there is no information available concerning the effects of CsA on urinary smooth muscle. Therefore, the effects of CsA were investigated on the bladder and urethral smooth muscle in rabbits. METHODS: CsA (10 mg/kg/day, intravenously) was given to rabbits for 14 days. Rabbit bladder and urethral smooth muscles were then isolated and evaluated using a muscle bath technique. RESULTS: 10(-10)-5 x 10(-6)M CsA alone had no direct effect on bladder or urethral smooth muscles. The Emax (maximum contractile response) and ED50 values for acetylcholine-induced contractions and the Emax (maximum relaxation response) for isoproterenol-induced relaxation in bladder smooth muscles were not significantly different between CsA-treated and control groups. The ED50 for isoproterenol-induced relaxation was significantly lower in the CsA group (P < 0.05). The Emax for phenylephrine- and clonidine-induced contractions in urethral smooth muscle was significantly higher in the CsA-treated group (P < 0.05). The ratio of the maximum response of the urethral smooth muscle to phenylephrine and clonidine in Ca+2-free solution to the normal solution in the CsA group (13.42% and 4.40%, respectively) was significantly higher than the maximal response ratios in the control group (6.34% and 3.00%, respectively; P < 0.05). CONCLUSIONS: CsA treatment augments the relaxation response of the bladder to isoproterenol and the contractile response of the urethra to phenylephrine and clonidine. In addition, CsA increases the filling of intracellular stores of releasable Ca+2, and also increases the permeability of Ca+2 in rabbit urethral smooth muscle. Thus, it is suggested that CsA may cause a urinary disturbance in patients treated with CsA via increased urethral resistance.

Acetylcholine↗

Subcapsular hematoma and hypertension following percutaneous needle biopsy of a transplanted kidney.

Percutaneous needle biopsy of a transplanted kidney is indicated as a diagnostic procedure in the evaluation of unexplained deteriorated renal function after renal transplantation. This procedure is not always without complications. We report a case of a subcapsular hematoma and hypertension following a percutaneous needle biopsy of a transplanted kidney. The cause of hypertension in this patient and the management of the subcapsular hematoma are discussed.

Adult↗

Establishment of two rat osteosarcoma cell lines (YROS-1 and YROS-2) induced by radioactive phosphorus.

Two rat osteosarcoma cell lines, YROS-1 and YROS-2, were established from two experimental osteosarcomas and induced by internal irradiation with radioactive phosphorus. Both cell lines formed a monolayer cell sheet in vitro with focal piling. The YROS-1 cells were refractile and spindle or polygonal in shape, whereas the YROS-2 cells were flat, spread and polygonal in shape. Ultrastructurally, the YROS-1 cells had well-developed rough-surfaced endoplasmic reticulum with focal pericellular deposition of calcified matrix, whereas YROS-2 had abundant polysomes and intracytoplasmic filaments. Both cell lines grew stably with population doubling times of 23 and 39 h, respectively. Flow cytometry revealed that YROS-1 was rich in proliferating cells compared to YROS-2, with a higher colony-forming efficiency. YROS-1 showed high alkaline phosphatase activity, while YROS-2 possessed low activity. When subcutaneously transplanted into lumbodorsal area of athymic nude mice, only YROS-1 formed tumors with frequent lung metastasis.

Animals↗

Seroepidemiology of hepatitis C virus infection in Japan and HCV infection in haemodialysis patients.

Since January 1990, Japanese Red Cross Blood Centres have introduced hepatitis C virus screening with a first-generation ELISA. From April to December 1992, approximately 0.98% among 10,905,489 blood donations screened by a second-generation assay were anti-HCV-positive in all Japan. Seropositivity of anti-HCV increased with the age and serum transaminase value in both sexes. In blood donors having a history of transfusion, the anti-HCV reactive rate was 7.4%. The results of the study made by the Japanese Red Cross Non-A, Non-B Hepatitis Research Group show the effectiveness of implementation of HCV screening to prevent posttransfusion hepatitis. Consecutive haemodialysis patients with chronic renal failure are at risk for infection by a variety of blood-borne agents transmitted within dialysis units. Because of their immunocompromised state, they frequently also have an unusual susceptibility to a variety of nosocomial infections, such as HBV, HCV, and HTLV-I. We tested the prevalence of anti-HCV in 1423 (848 males and 575 females) haemodialysis patients from 18 hospitals in Kumamoto Prefecture, Japan, using the Ortho first generation anti-HCV screening assay. There were 316 patients (22.2%) positive for HCV antibodies. The second-generation test was positive in most haemodialysis patients who were reactive to the first-generation assay. The prevalence of HCV infection increased with the duration of haemodialysis, yet there was a high frequency of HCV seropositivity even without blood transfusion. Acquisition of HCV in dialysis patients could be explained by HCV infection within the unit other than by blood (all haemodialysis are done with disposable kits, syringes, and needles), by secondary HCV infection after the immunodeficiency of haemodialysis, or by HCV infection of the kidney or glomerular deposition of immune HCV/anti-HCV complexes leading to chronic renal failure (as with HBV infection of the liver and kidney.

Blood Donors↗

Vascular response to vasoactive agents in dialysed patients with chronic renal failure with and without diabetes mellitus.

Using a muscle bath technique the vascular response to KCl, noradrenaline, angiotensin II, acetylcholine, and sodium nitroprusside were evaluated in 13 patients with diabetes mellitus (DM group) and 15 nondiabetic (non-DM group) chronic renal failure patients treated with haemodialysis. There were no differences in age, duration of haemodialysis, blood pressure and the levels of plasma renin activity, noradrenaline, and parathyroid hormone between groups. After informed consent was obtained, a small piece of forearm vein was resected during the blood access surgery. The ring preparation of the blood vessel was sustained in the muscle bath filled with Krebs-Henseleit solution and the isometric tension development was recorded. All drugs produced concentration dependent responses in the ring preparations of both groups. Although there were no significant differences in Emax values for KCl- and angiotensin-II-induced contractions between groups, the value for noradrenaline in the DM group was significantly less than that in the non-DM group. Sodium nitroprusside completely relaxed the ring preparation precontracted by 10(-5) M noradrenaline. However, the response to acetylcholine in the DM group was significantly weaker than that in the non-DM group. These results suggest a reduced vascular response to noradrenaline and acetylcholine in dialysed diabetic renal failure patients, which may relate to the autonomic nervous system dysfunction.

Adult↗

Masticatory function in a patient with mandibular midline cleft.

Bone grafting of a midline cleft is required to allow reasonable masticatory function. However, changes in the rhythm of mastication have not been investigated before and after such bone grafting procedures. We performed bone grafting in a patient with multiple craniofacial anomalies including a midline mandibular cleft. The patient showed a marked improvement in the rhythm of mastication after repair of the midline mandibular defect. Bone grafting proved to be an effective method for securing stable occlusion with better mandibular movement.

Bone Transplantation↗

Grading system of articular cartilage degeneration in osteoarthritis of the knee.

Degenerative findings observed in the articular cartilage of the knee during high tibial osteotomy with arthrotomy, unicompartmental arthroplasty, or total replacement were classified into 9 grades: Grade 0--normal, Grade I--yellow discoloration, Grade II--softening and unevenness, Grade III--fasciculation and attrition, Grade IV-a--erosion, Grade IV-b--ulcer down to subchondral bone, Grade IV-c--eburnation of subchondral bone, Grade V-a--bone destruction in an area less than 5 mm in depth, and Grade V-b--bone destruction in an area 5 mm or more. This series included 63 osteoarthritic knees with varus deformity in 56 Japanese patients (forty nine women and seven men) with an average age of 66.8 years (range: 49-83 years). One knee was Grade II, three were Grade III, ten Grade IV-a, eighteen Grade IV-b, twenty seven Grade IV-c, and four Grade V-b. There was a tendency toward positive correlation between the grade of articular cartilage degeneration and the radiographic grade seen in a standing radiograph.

Aged↗

Effects of phorbol ester on lower urinary tract smooth muscles in rabbits.

The contractile effects of phorbol 12,13-dibutyrate (PDBu) on rabbit urinary bladder dome and urethra were investigated using muscle bath techniques. PDBu caused concentration-dependent contractions in both tissues, and these responses were not affected by pretreatment with atropine, phentolamine, hexamethonium or indomethacin. In both tissues, 1-(5-isoquinolinyl-sulfonyl)-2-methylpiperazine (H-7), a potent inhibitor of protein kinase C (PKC), inhibited PDBu-induced contractions in a concentration-dependent manner. The maximum PDBu-induced contractions in bladder dome and urethra were 33.5 +/- 3.4 and 33.3 +/- 4.1% of KCl-induced maximum contractions, respectively. In Ca(2+)-free solution or after pretreatment with nifedipine (10(-6) M), PDBu-induced contractions were reduced but not completely abolished. Although pretreatment with PDBu (10(-8) M) did not have a significant effect on the contractile responses induced by carbachol (in bladder dome) and phenylephrine (in urethra), pretreatment with H-7 (100 microM) had an inhibitory effect on carbachol- and phenylephrine-induced contractions; tonic phase contractions were more sensitive than phasic contractions. These results indicate that PDBu has significant contractile effects in rabbit bladder dome and urethra, and that the effects may be partly mediated by activation of PKC. PKC activation might also contribute to agonist-induced contractile responses in these tissues.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗