Bilateral Kienbock's disease.

We conducted a retrospective review of 11 patients with bilateral Kienbock's disease from our series of 251 patients with Kienbock's disease. There were no significant differences in radiographic parameters, including ulnar variance and carpal bone angle, between those with unilateral and those with bilateral Kienbock's disease. None of the patients with bilateral disease had been treated with corticosteroids or had a systemic disease that predisposed to osteonecrosis. Thus, this study failed to demonstrate any risk factor for bilateral, as opposed to unilateral Kienbock's disease.

Radiographic morphology of the ulnar head.

Posteroanterior radiographs of 102 normal wrists were studied to determine the morphological characteristics of the ulnar head associated with the value of ulnar variance. Ulnar variance, ulnar styloid length, ulnar head diameter, ulnar seat inclination, and the distance of ulnar head peak to the distal radioulnar joint were measured together with the third metacarpal length as a reference of the size of the wrist. Moderate correlation was confirmed not only between ulnar variance and ulnar seat inclination but also between ulnar variance and the distance of the ulnar head peak. Furthermore, we found a statistically significant correlation between ulnar variance and ulnar head diameter. The results showed that whole ulnar head configurations are affected by ulnar variance although there are considerable variations.

Inhibition of mutant p53 phosphorylation at serine 15 or serine 315 partially restores the function of wild-type p53.

The tumor suppressor protein p53 is a phosphoprotein and has growth and transformation suppression functions. Phosphorylation of wild-type p53 is known to modulate its function. To investigate the role of phosphorylation in modulating the functions of mutant p53, we constructed a series of phosphorylation site mutants based on mutant p53 Ala143 (p53-143) and p53 His175 (p53-175). When transfected into p53-negative Saos-2 cells, parental mutant p53-143 and p53-175 abolished both growth suppression and induction of apoptosis. However, DNA-activated protein kinase (DNA-PK) or cyclin-dependent kinase (cdks) phosphorylation site double mutants partially restored the growth suppression and induction of apoptosis and recovered the p53-specific DNA binding activity. We also observed a difference in sensitivity to calpain from parental mutants p53-175 and p53-175/15 or p53-175/315. These results suggest that the lack of phosphorylation at either the DNA-PK or cdks site in p53 mutants partially restores the wild-type functions by altering their conformation.

Mutant p53 mediated induction of cell cycle arrest and apoptosis at G1 phase by 9-hydroxyellipticine.

Wild-type p53 causes cell-cycle arrest at late G1 phase and induction of apoptosis by up- regulation of waf1 and bax, respectively. Although in many cancer cells p53 is frequently mutated and loses its functions, we have proposed that mutant p53 may be involved in the anticancer mechanism of 9-hydroxy ellipticine (9HE). It was shown using flow cytometry that 9HE (10 microM) caused induction of apoptosis in G1 phase of the cell cycle in mutant p53 (p53ala143, p53his175, orp53his273) transfected Saos-2 cells, but not in p53-deficient parental Saos-2 cells. Similar induction of apoptosis was observed 24-48 h after treatment with 9HE in mutant p53-containing SW480, SK-BR-3 and MKN-1, but not in p53-deficient KATO m cells. Using G1 phase cells isolated by centrifugal elutriation, it was confirmed that 9HE caused cell cycle arrest at G1 phase and subsequently induced G1 phase-restricted apoptosis. In accordance with the induction of arrest and apoptosis in G1 phase, 9HE caused up-regulation of waf1 and bax mRNA in mutant p53-containing cells, but not in p53-deficient parental Saos-2 cells. In control experiments, adriamycin (ADR) showed neither G1-restricted apoptosis nor elevation of bax mRNA. It is suggested that 9HE may cause G1 arrest and induction of G1 phase-restricted apoptosis by restoring the wild-type function of mutant p53 protein.

Severe bleeding tendency in a patient with Bernard-Soulier syndrome associated with a homozygous single base pair deletion in the gene coding for the human platelet glycoprotein Ibalpha.

PURPOSE: The genetic basis of Bernard-Soulier syndrome (BSS) was studied to clarify a relationship between severe clinical manifestations and gene abnormality. PATIENT AND METHODS: A patient with BSS had a severe bleeding tendency that was sometimes life threatening. Flow cytometric analysis of the patient's and normal control platelets was performed to study which glycoprotein (GP) was impaired in glycoprotein Ib/V/IX complex. The genes encoding GPIbalpha from the patient's and control genomic DNA were amplified and directly sequenced. RESULTS: Flow cytometric analysis revealed a defect of GPIbalpha on the surface of the patient's platelets. A homozygous single base pair deletion was identified in seven repeats of adenine at positions 1932 to 1938 in the GPIbalpha gene. This mutation, which has been previously reported, results in a frameshift and predicts a premature stop codon leading to a truncated peptide that cannot fix on the platelet membrane. CONCLUSION: This patient's severe clinical phenotype would be explained by this mutation in the GPIbalpha gene.

Pharmacological profile of T-0632, a novel potent and selective CCKA receptor antagonist, in vivo.

The pharmacological profile of a new CCKA receptor antagonist, T-0632 [sodium (S)-1-(2-fluorophenyl)-2,3-dihydro-3-[(3-isoquinolinylcarbonyl) amino]-6-methoxy-2-oxo-1H-indole-3-propanoate], was examined in in vivo studies and compared with those of L-364, 718 [3S(-)-N-(2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1, 4-benzodiazepine-3-yl)-1 H-indole-2-carboxamide] and loxiglumide [D.L-4-(3,4-dichlorobenzoylamino)-5-(N-3-methoxypropyl-pentylam ino)-5- oxopentanoic acid]. In rats, intravenously administered T-0632, L-364,718 and loxiglumide dose dependently inhibited cholecystokinin octapeptide (CCK-8)-stimulated pancreatic exocrine secretion with estimated ED50 values of 0.025, 0.016 and 1.8 mg/kg, respectively. The ED50 values for intraduodenal administration of these compounds were 0.040, 0.26 and 3.0 mg/kg, respectively. In mice, orally administered T-0632 prevented caerulein-induced pancreatitis, CCK-8-induced inhibition of gastric emptying and CCK-8-induced gallbladder emptying in dose-dependent manners with ED50 values of 0.028, 0.04, and 0.12 mg/kg, respectively. The effect of T-0632 for caerulein-induced pancreatitis was 4-fold more potent than that for gallbladder emptying. In contrast, the effects of L-364,718 and loxiglumide for caerulein-induced pancreatitis were 2-4-fold weaker than those for gallbladder emptying. In dogs, T-0632 and loxiglumide maximally inhibited CCK-8-stimulated pancreatic amylase secretion at doses of 0.01 and 10 mg/kg, respectively. At these doses, the effect of T-0632 on CCK-8-induced increase in the gallbladder intraluminal pressure was weaker than that of loxiglumide. These results suggest that T-0632 has a potent antagonistic action on CCKA receptors in several animal species and the effects of T-0632 are more selective for the pancreas over the gallbladder compared with L-364,718 and loxiglumide.

Pharmacological profile of T-0632, a novel potent and selective CCKA receptor antagonist, in vitro.

The pharmacological profile of a new CCKA receptor antagonist, T-0632 [sodium (S)-3-[1-(2-fluorophenyl)-2,3-dihydro-3-[(3-isoquinolinyl)-carbonyl] amino-6-methoxy-2-oxo-1-H-indole]propanoate], was examined in in vitro studies and compared with those of L-364,718 [3S(-)-N-(2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1, 4-benzodiazepine-3-yl)-1H-indole-2-carboxamide] and loxiglumide [D,L-4-(3,4-dichlorobenzoylamino)-5-(N-3-methoxypropyl-pentylam ino)-5- oxopentanoic acid]. T-0632 inhibited the specific binding of [125I]CCK-8 to rat pancreatic CCKA receptor in a concentration-dependent and competitive manner. The Ki value of T-0632 for the CCKA receptor was estimated to be 0.24 nM, which was 23 000-fold less than the Ki value (5600 nM) for guinea pig CCKB receptor. L-364,718 and loxiglumide were 1500- and 64-fold selective for CCKA over CCKB receptor, respectively. T-0632, L-364,718 and loxiglumide inhibited CCK-8 (100 pM)-stimulated amylase release from rat pancreatic acini in a concentration-dependent manner with IC50 values of 5.0 nM, 5.0 nM and 3.0 microM, respectively. In the isolated rabbit gallbladder smooth muscle, T-0632 and loxiglumide competitively inhibited CCK-8-induced contraction with pA2 values of 8.5 and 7.0, respectively. However, L-364,718 showed an apparent non-competitive antagonism. The IC50 values of T-0632, L-364,718 and loxiglumide for CCK-8 (30 nM)-induced contraction were 31 nM, 4.9 nM and 1300 nM, respectively. The inhibitory effects of T-0632 and loxiglumide in gallbladder smooth muscle were readily reversible, but L-364,718 showed a long-lasting inhibition. These results suggest that T-0632 is a potent, reversible and more selective CCKA receptor antagonist compared with L-364,718 and loxiglumide.

Longitudinal investigation of epidemiologic feature of adenovirus infections in acute respiratory illnesses among children in Yamagata, Japan (1986-1991).

Using human embryonic fibroblast (HEF) and HEp-2 cell cultures, adenoviruses were isolated from 989 (3.7%) out of 26,793 pediatric patients with ARI in Yamagata, Japan from January, 1986 to December, 1991. All isolates were identified as types 1 (Ad1)-6 and no other serotypes were identified. Epidemiologic feature was different depending on the subgenus group. Ad1, 2, 5 and 6 (group C) were endemic and the infections occurred frequently in the summer season. Ad3 (group B) was epidemic in the autumn to winter season, although the virus was isolated every month in non-epidemic season. No seasonal distribution of Ad4 (group E) could be determined because the number of patients was limited. Neutralizing antibody positive ratio for group C were more than 40% at 1-2 years of age and almost 100% by 10 years of age but those for Ad3 (group B) were 40% by 10 years of age. The neutralizing antibodies for Ad4 (group E) or Ad7 (group B) became negative by 10 years of age. With group C infections, most cases were infants and young children less than 2 years of age, but Ad3 infections were older children with the peak at 4 and 5 years of age.

Differential expression patterns of mRNAs for members of the fibroblast growth factor receptor family, FGFR-1-FGFR-4, in rat brain.

We have examined the region-specific expression of mRNAs for four members of rat FGF receptor family, FGFR-1, FGFR-2 FGFR-3, and FGFR-4, in rat brain by in situ hybridization. The FGFR-1, FGFR-2, and FGFR-3 mRNAs were expressed widely but differentially in the brain. However, the FGFR-4 mRNA was not expressed in the brain. The FGFR-1 mRNA was strongly expressed in several regions including the hippocampus, cerebellum, and pedunculopotine tegmental nucleus. The FGFR-2 mRNA expression was high in the choroid plexus, and moderate in the fiber-rich regions (the corpus callosum, external capsule, and internal capsule) and the olfactory bulb. The FGFR-3 mRNA was expressed diffusely in the brain. We have also examined the cellular localization of these mRNAs in the brain. Although the FGFR-1 mRNA was expressed preferentially in neurons, the FGFR-2 and FGFR-3 mRNAs were expressed preferentially in glial cells. The present findings that the FGFR-1, FGFR-2, and FGFR-3 mRNAs were expressed widely but with region- and cell-specificity in the brain indicate that these receptors have different roles in the brain.

Rat FGF receptor-4 mRNA in the brain is expressed preferentially in the medial habenular nucleus.

The fibroblast growth factor (FGF) receptor family consists of four members, FGFR-1, FGFR-2, FGFR-3 and FGFR-4, that are closely related receptor tyrosine kinases. We examined the expression of rat FGFR-4 mRNA in the brain by in situ hybridization and compared it with that of the mRNAs for other FGF receptors. In contrast with FGFR-1, FGFR-2 and FGFR-3 mRNAs which are expressed widely in the brain, the FGFR-4 mRNA in the brain is expressed preferentially in the medial habenular nucleus neurons. The present finding indicates that FGFR-4 has a function specific to the medial habenular nucleus.

Six-year longitudinal analysis of adenovirus type 3 genome types isolated in Yamagata, Japan.

Five hundred eighty-seven adenovirus type 3 (Ad3) isolates were established from children with acute respiratory infections (ARI) from 1986 to 1991, in Yamagata, Japan. Ad3 could be found in almost all the months during the 6 years when two epidemics occurred, in 1987 and 1989. A molecular epidemiological study was done on 346 of the 587 isolates, using restriction endonucleases; BamHI, HindIII, SmaI, and BgIII were used. The Ad3 isolates were classified into seven genome types. The genetic differences among the seven genome types were < 0.9%, and their phylogenetic tree, estimated by the neighbor-joining method, correlated highly with their monthly distribution. One genome type predominated for 56 months, while the other six related genome types cocirculated for a short period. These results suggested that the predominant genome type of Ad3 might have been endemically perpetuated in the Yamagata area with minor genomic variations. Furthermore, the outbreaks of Ad3 may have been due not to the appearance of a new genome type but rather to the endemic genome type.

The structure and expression of the FGF receptor-1 mRNA isoforms in rat tissues.

In this paper, we describe the structure of rat FGF receptor-1 mRNA isoforms and their expression in a variety of rat tissues. The rat FGFR-1 has the characteristics of FGFR-1 as well as mouse, human and chicken homologs. FGFR-1 mRNA was detectable in all the tissues examined by Northern analysis or polymerase chain reaction, indicating that FGFR-1 mRNA is widely expressed in rat tissues. The rat FGFR-1 mRNA has isoforms in both the extracellular and intracellular regions. The extracellular isoforms which have two or three immunoglobulin-like domains, are expressed almost equally in the tissues except for brain. However, the large form is a major form in the brain. Furthermore, in the brain, half of FGFR-1 mRNAs have the six nucleotides, which encode a potential serine-threonine kinase phosphorylation site in the intracellular juxta-membrane region, deleted. In contrast to the brain, the deletion isoform is a minor form in the other tissues. The tissue-specific expression of the isoforms indicates that they have different physiological functions. Although other isoforms of FGFR-1 mRNA in tumor cell lines have been reported, the isoforms were undetectable in all rat tissues examined, indicating the isoforms are products of abnormal alternative splicing in tumor cell lines.

Regression in schizophrenia and its therapeutic value.

Using the regression evaluation scale, 25 schizophrenic patients were classified into three groups of Dissolution/autism (DAUG), Dissolution----attachment (DATG) and Non-regression (NRG). The regression of DAUG was of the type in which autism occurred when destructiveness emerged, while the regression of DATG was of the type in which attachment occurred when destructiveness emerged. This suggests that the regressive phenomena are an actualized form of the approach complex. In order to determine the factors distinguishing these two groups, I investigated psychiatric symptoms, mother-child relationships, premorbid personalities and therapeutic interventions. I believe that these factors form a continuity in which they interrelatedly determine the regressive state. Foremost among them, I stressed the importance of the mother-child relationship.

[Systolic time intervals in mitral valve prolapse syndrome].

Systolic time intervals (STI) were measured in 135 patients with mitral valve prolapse syndrome (MVP). These patients were categorized as Group II (no or trivial mitral regurgitation) and Group III (moderate or severe regurgitation). The controls consisted of 120 normal subjects (Group I). The Group II patients tended to have increased ETc and shortened PEP. The Group III patients tended to have prolonged Q-I and decreased QIIc and ETc, reflecting mitral regurgitation. It was suggested that autonomic nervous system, especially sympathetic nerve, may play a role in changes of STI of the Group II patients.

Effect and follow-up study on varicella vaccine.

Attenuated liver varicella vaccine (Oka strain) was used to vaccinate 242 children and 5 adults between August 1976 and December 1982; namely emergency vaccinations were given to 163 cases, including 35 high risk children, on 17 occasions, and non-emergency vaccinations were given to 84 cases including 7 high risk ones in remission. The viral doses varied from 250 to 3,000 PFU. Vaccinations prevented subsequent infection in all cases. Emergency vaccinations were given within 100 h after contact of the subjects with cases of varicella. Humoral and/or cellular immunity was acquired in 97.6% (40/41) of the high risk group and 91.8% (179/195) of the non-high risk group. As clinical reactions, rashes and fever developed in 43.9% (18/41) and in 17.0% (7/41) of high risk patients, and 7.8% (16/204) and 1.0% (2/204) of the non-high risk patients respectively. Reactions were generally slight, but were severe or atypical in 3 immunocompromized patients. Follow-up studies were carried out every year since 1980. Among the 41 high risk patients, herpes-zoster developed in 4, and varicella in 5 patients. Among the 179 non-high risk patients, there were no cases of herpes-zoster but 21 cases (12.3%) of varicella, which were mostly extremely mild. Six patients were revaccinated because of their humoral and/or cellular immunity decreased, and as a result acquired an immune response again. Criteria for varicella vaccination and details of the results of vaccination and follow-up studies are described.

Changing RNA patterns in rotaviruses of human origin: demonstration of a single dominant pattern at the start of an epidemic and various patterns thereafter.

The RNA of strains of rotavirus obtained from patients hospitalized with diarrhea during two winter epidemics of rotaviral infection in successive years (November 1981 through April 1982 and December 1982 through April 1983) was analyzed by polyacrylamide gel electrophoresis. A single dominant electropherotype was found during the first two or three months of each epidemic. In contrast, various electropherotypes were identified during the latter portion of each epidemic. RNA patterns of the rotaviral strains that were dominant during the early phase of the two epidemics were different from each other.