Genetic epidemiology of tooth agenesis in Japan: a population- and family-based study.

Tooth agenesis is one of the most common congenital anomalies in humans. However, the etiology of tooth agenesis remains largely unclear, as well as evidence base useful for genetic counseling. Therefore, we estimated the prevalence and sibling recurrence risk, and investigated agenetic patterns systematically. Tooth agenesis was classified into two subtypes: hypodontia (one to five missing teeth) and oligodontia (six or more missing teeth). The prevalence of these two subtypes were 6.8% [95% confidence interval (CI): 6.1-7.7%] and 0.1% (95% CI: 0.04-0.3%), respectively, and sibling recurrence risk of these were 24.5% (95% CI: 13.8-38.3%) and 43.8% (95% CI: 26.4-62.3%), respectively. This result suggests that the severe phenotype, oligodontia, might be mostly transmitted in a dominant fashion. Using a simple statistical modeling approach, our data were found to be consistent with a bilateral symmetry model, meaning that there was equal probability of missing teeth from the right and left sides.

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.

New radiological parameters for the assessment of atlantoaxial instability in children with Down syndrome: the normal values and the risk of spinal cord injury.

AIMS: To determine the normal values and usefulness of the C1/4 space available for spinal cord (SAC) ratio and C1 inclination angle, which are new radiological parameters for assessing atlantoaxial instability in children with Down syndrome. PATIENTS AND METHODS: We recruited 272 children with Down syndrome (including 14 who underwent surgical treatment), and 141 children in the control group. All were aged between two and 11 years. The C1/4 SAC ratio, C1 inclination angle, atlas-dens interval (ADI), and SAC were measured in those with Down syndrome, and the C1/4 SAC ratio and C1 inclination angle were measured in the control group. RESULTS: The mean C1/4 SAC ratio in those requiring surgery with Down syndrome, those with Down syndrome not requiring surgery and controls were 0.63 (standard deviation (sd) 0.1), 1.15 (sd 0.13) and 1.29 (sd 0.14), respectively, and the mean C1 inclination angles were -3.1° (sd 10.7°), 15.8° (sd 7.3) and 17.2° (sd 7.3), in these three groups, respectively. The mean ADI and SAC in those with Down syndrome requiring surgery and those with Down syndrome not requiring surgery were 9.8 mm (sd 2.8) and 4.3 mm (sd 1.0), and 11.1 mm (sd 2.6) and 18.5 mm (sd 2.4), respectively. CONCLUSION: The normal values of the C1/4 SAC ratio and the C1 inclination angle were found to be about 1.2° and 15º, respectively. Cite this article: Bone Joint J 2016;98-B:1704-10.

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.

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.

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.

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.

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.

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.

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.

[Renovascular hypertension. Changes and prognosis of the surgical treatment].

We investigated 663 patients with renovascular hypertension (349 males and 314 females), who were surgically treated at 73 institutions of Urology or Surgery in Japan from 1963 to 1988. There were 294 cases with fibromuscular dysplasia (FMD), 178 with atherosclerosis (SC), 104 with aortitis syndrome (AO) and 87 with other causes. The mean age was 27 years old for FMD, 52 years for SC and 29 years for AO. Thirteen % of FMD, 19% of SC and 38% of AO had stenoses in both renal arteries. Since percutaneous transluminal angioplasty (PTA) has begun in early 1980's, the number of operative cases has been decreased, especially in the case of FMD due to a good result by PTA. Nephrectomy was done in 38% of 704 operations. Aortorenal bypass and autotransplantation were performed in 49% and 26% of the rest, respectively. Recently, autotransplantation has increasingly been applied for complicated vascular lesions. As regards prognosis of the surgical treatment, 77% of FMD, 59% of SC and 50% of AO presented normal blood pressure with or without antihypertensive drugs at least for 10 years after an operation. However, among the patients with normal blood pressure who were observed for more than 5 years, 30% of FMD, 56% of SC and 44% of AO had taken antihypertensive drugs. Eleven patients (1.7% of all) died within one month after the operation, 5 patient of whom had bilateral stenoses. In conclusion, PTA is the first choice for renovascular hypertension and surgery has been done for more difficult and complicated cases in which PTA is ineffective.(ABSTRACT TRUNCATED AT 250 WORDS)

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.

Mutation of keratin 4 gene causing white sponge nevus in a Japanese family.

White sponge nevus (WSN) is a rare autosomal dominant disorder characterized by white plaques of oral mucosa; it is benign condition with no effective treatment. The disorder usually manifests during early childhood or adolescence. Mutations of keratin 4 or 13 gene have been identified as causing WSN. The aim of this study is to determine whether keratin 4 or 13 gene mutation was the molecular basis of WSN in a Japanese family. The proband in this family was an 11-year-old boy, with three other people affected by WSN. Genomic DNA was extracted from two affected members and an unaffected member. Segments of keratin 4 and 13 genes were amplified by PCR, and direct DNA sequencing was carried out. Sequence analysis revealed a heterozygous 3bp deletion (N160Del) localized in the helix-initiation motif at the beginning of alpha-helical domain 1A of keratin 4 gene from affected members. One member lacking the phenotype was genetically tested normal. The authors identified a mutation of the keratin 4 gene recurrent in a family affected by WSN. Further investigation of the multifunctional role of keratin genes is warranted in the group of inherited epithelial disorders that may result in identification of effective treatment for this genetic disease.

A novel mutation in the C-terminal region of RUNX2/CBFA1 distal to the DNA-binding runt domain in a Japanese patient with cleidocranial dysplasia.

Cleidocranial dysplasia (CCD) is an autosomal dominant inherited skeletal disease with high penetrance and variable expressivity. Although many mutations in RUNX2/CBFA1, an osteoblast-specific transcription factor, have been identified as causes of CCD, it is unclear whether these mutation genotypes relate to various symptoms. Heterogeneous mutations of RUNX2/CBFA1 result in disease characterized by abnormal skeletal genesis and dental disorders. There are few reports describing the relation between detailed orofacial pathology and the RUNX2/CBFA1 genotype. The case of a Japanese patient with severe orofacial dysplasia who was clinically thought to have CCD is described here. The authors performed mutation analysis on the RUNX2/CBFA1 gene and identified a novel frameshift mutation (722delT), which produces a mutant RUNX2/CBFA1 with a truncating C-terminus distal to the runt domain.

Massive immature teratoma in a neonate.

Teratomas are embryonal tumours composed of diverse tissues from three germinal layers with variable levels of maturity. The authors report a female patient with a large immature epignathus. Prenatal diagnosis permitted a caesarean section and tracheotomy to be planned under ex utero intrapartum treatment for airway obstruction. The tumour was successfully controlled, even though it was impossible to distinguish from normal tissue because it lacked a pedicle and capsule, using multidisciplinary therapy, including a series of surgical treatments and adjuvant chemotherapy. This case suggested that the level of serum alpha fetoprotein might be a useful indicator after surgery. At present, after 4 years, no regrowth has been observed and the patient has no problems with respiration or swallowing.