Preparation of monoclonal antibodies to chicken bone phosphoproteins

Summary Monoclonal and polyclonal antibodies were raised against 150k, 60k, 32k, and 15k phosphoprotein components of 14-week chicken bone. Hybridomas were prepared by immunizing Balb/c mice with 150k, 60k, or 32k phosphoproteins followed by fusion of their spleen cells with X63-Ag8.653 myeloma cells. Polyclonal antibodies to the 60k, 32k, and 15k phosphoprotein components were produced in immunized rabbits. Immunological cross-reactivity between antigen and antibody was examined by enzyme-linked immunosorbent assay (ELISA) and dot-blotting. There was cross-reactivity between the 150k, 32k, and 15k phosphoprotein components and between the 150k, 90K, and 60k components. The antibodies raised against 60k component do not bind 32k and 15k antigens.     

Relationship between cell proliferation activity and morphological characteristics of papillary microcarcinoma in the thyroid of Graves' disease

Three hundred and seven lesions of papillary microcarcinoma of the thyroid ( PMT ), found in 6830 cases of Graves' disease, were reviewed to evaluate the morphological characteristics. The prevalence rate and incidence of multiplicity of PMT in Graves' disease were higher ( P < 0.05 and P < 0.005, respectively) in females (4.03% and 0.73%, respectively) than in males (2.68% and 0.12%, respectively). All lesions of PMT were classified histologically into three types: sclerosing non-encapsulated (SNE), non-sclerosing encapsulated (NSE) and non-sclerosing nan-encapsulated (NSNE). Comparison of the three types of PUT revealed that the mean age and tumor size of NSNE were less than those of NSE and SNE (P < 0.001 and P < 0.005, respectively), and 13 of 19 lesions less than 1 mm belonged to NSNE. In addition, cell proliferation and expression of estrogen and progesterone receptors were examined immunohisto-chemically in 257 lesions of PMT. The Ki-67 labeling index of NSE was lower than that of NSNE and SNE ( P < 0.005). None of the cases of PMT was positive for estrogen or progesterone receptors. The present study indicated that NSNE may be an early stage in tumor progression, and that fibrous encapsulation has the potential to modify neoplastic cell proliferation.     

Age-dependent Alteration of the Serum-unbound Fraction of Nicardipine,a Calcium-channel Blocker,in Man

To determine whether the age-dependent increase in the pharmacological effect of calcium-channel blockers is a result of age-dependent alteration of the unbound fraction the drug in serum, the unbound fraction of the nicardipine was investigated in the serum of 38 adults. The unbound concentration of nicardipine in serum to which nicardipine (205.4 ng mL^?1) had been added was determined by ultracentrifugation to range from 0.49 to 4.01% (mean±s.d., 1.55 ± 0.78%). Non-glycosylated albumin was most strongly correlated with age (r = 0.901). Total bilirubin was weakly correlated with age whereas levels of α-1-acid glycoprotein, triglycerides and glycosylated albumin were not correlated with age. A significant (P < 0.01) linear correlation was obtained between the unbound fraction of nicardipine and parameters such as age, albumin, albumin/globulin ratio, albumin/glycosylated albumin ratio, non-glycosylated albumin and total bilirubin. To assess the relative effect of each variable on the unbound fraction of nicardipine, stepwise multiple linear regression was performed using age and biochemical parameters. The three variables (non-glycosylated albumin, total bilirubin and age) were entered into the regression equation. The results of this study showed that the major ligand of nicardipine in serum was non-glycosylated albumin, which decreased with age. It was, moreover, shown that the serum-unbound concentration of nicardipine increased with age. This finding would be one factor accounting for the increase in the pharmacological effect of nicardipine with age. In addition, our predicted model for the unbound fraction of nicardipine might be useful in determining the appropriate nicardipine dose for the elderly.     

Reliability and validity of the Childhood Autism Rating Scale-Tokyo version (CARS-TV)

A Japanese translation of the Childhood Autism Rating Scale (CARS) (the Tokyo version of the CARS, CARS-TV) was used with 167 developmentally disabled children under age 16. Cronbach's coefficient alpha was 87. The interrater reliability (r) for each of the 15 scales based on 128 children ranged from .43 to .77 with an average of .62. Based on the 167 children, the total CARS-TV score demonstrated a satisfactory level of taxonomic validity (Thorndike, 1982) on DSM-III diagnostic groups. The total score discriminated infantile autism and other pervasive developmental disorders more efficiently from mental retardation without an additional diagnosis of pervasive developmental disorder than an IQ. The total score also showed a satisfactory concurrent validity on the overall rating of autism.This study was supported in part by a grant from the Ministry of Education, Science and Culture of Japan (No. 63480263). The authors thank Dr. E. Schopler for his permission to use a Japanese translation of the CARS in this study.     

Characteristics of antinociception induced by noncatecholic phenylethylamine derivatives: the relation of endogenous norepinephrine to phenylethylamine analog-induced antinociception

Characteristics of the antinociceptive action of phenylethylamine derivatives, amphetamine, beta-phenylethylamine (PEA) and beta-hydroxyphenylethylamine (OHPEA), were examined. The pain threshold of mice was measured by using the hot plate method. Intraperitoneal administration of alpha-methyl-p-tyrosine inhibited antinociception induced by PEA and OHPEA, and intracisternal administration of norepinephrine increased antinociception induced by PEA and OHPEA. Intracisternal administration of phentolamine inhibited the antinociception induced by PEA derivatives. The levels of norepinephrine and normetanephrine in the brain were determined by using HPLC. PEA derivatives decreased norepinephrine in the brain and tended to increase normetanephrine at 15 min after the administration of PEA derivatives. These findings indicate that PEA derivatives cause the release of norepinephrine in the central nervous system, and the released norepinephrine induces antinociception.     

Plasma Cyclic AMP Level in Psychiatric Diseases of Childhood

Abstract: Determinations were made of the plasma cyclic AMP level to examine its relationship with hyperkinesis (Werry-Weiss-Peters Activity Scale, WWPAS) and other features of mental disorders in 80 children, of whom 21 had early infantile autism, 15 hyperkinetic mental retardation, 12 minimal brain dysfunction and 32 Down's syndrome. In autistic and hyperkinetic mentally retarded children, the plasma cyclic AMP levels were higher than in normal children and were positively correlated with the WWPAS score. In children with minimal brain dysfunction, the plasma cyclic AMP level was significantly lower than in normal children and was not correlated with the WWPAS score. In children with Down's syndrome, the plasma cyclic AMP level was somewhat higher than in normal children.     

Brachial plexus injuries. Prognosis of postganglionic lesions

One hundred ninety-eight cases of postganglionic brachial plexus injury with no signs of combined damage to preganglionic sites were treated conservatively. Among them, 50 whole, 32 upper and 13 lower type injuries were followed up for at least 2 years and the course of the progression of the individual muscle was observed. In the whole type, 44% showed good functional recovery, although some of them needed reconstructive surgery. Thirty-eight percent recovered usefully only in the muscles innervated by upper roots. Three out of 50 cases did not recover at all. There were no particular differences between the whole and upper types with regard to the recovery rate of the arm muscles. However, the upper type showed better functional recovery than the whole type. If the muscles innervated by upper roots could gain a strength of M1 before 9 months after injury, and muscles innervated by lower roots reached M1 before 12 months, their final results were more than M3. But muscles with a strength of less than M2 at 18 months did not recover to more than M3. The muscular power at 24 months was almost the same as the final results. About half of the postganglionic brachial plexus injuries which did not combine preganglionic lesions showed favorable spontaneous recovery with conservative treatment. At the exploration of the brachial plexus, if the somatosensory evoked potential test is positive, which negates the possibility of root avulsion, and also if there is no definite discontinuity of the postganglionic nerves, it should be treated conservatively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Changes in polypeptide pattern of rat liver cells during chemical hepatocarcinogenesis

Administration of 2-acetylaminofluorene to rats for 12 weeks induces hyperplastic nodules (HPNs) and later well-differentiated hepatocellular carcinomas (HCCs) in the liver. Total cellular proteins from normal liver, HPN, and HCC were analyzed by two-dimensional gel electrophoresis with a high resolution. Several hundred polypeptides were well resolved as seen by Coomassie blue staining, forming a reproducible and characteristic pattern for each tissue. The polypeptide patterns were very similar among normal liver, HPN, and HCC. Especially the proteins of HPN and HCC were almost indistinguishable. These neoplastic lesions, however, were clearly different from control liver in that a new spot p35-6.6 (designated by molecular weight X 10(-3) and pl) appeared, and five polypeptides, p57-6.9, p57-6.7, p26-6.9, p26-6.6, p26-6.4, increased dramatically in amount as compared with normal liver. These last three spots were found to be a new type of glutathione S-transferase as judged from the specific binding to the antibody. The same changes in polypeptide pattern were found in HCCs induced by other chemical carcinogens, diethylnitrosamine and 3'-methyl-4-dimethylaminoazobenzene, but not in regenerating and neonatal livers. Fetal liver showed a rather different pattern than adult liver, but only p26-6.6 was increased among the spots characteristic of HPN and HCC. Protein phosphorylation was also examined for these cells by incubating tissue slices with 32PO4. After alkali treatment of the gels to eliminate serines phosphorylation, several dozens of phosphoproteins were clearly detected. The patterns of the labeled spots were again very similar among control liver, HPN, and HCC. Only the intensity of a spot designated p57-6.6 increased markedly in both HPN and HCC. This spot was further resolved by an expanded pH gradient into four distinct spots, the major one of which contained phosphothreonine. Similar changes in phosphorylation were noted in hepatomas induced by diethylnitrosamine and 3'-methyl-4-dimethylaminoazobenzene but not in regenerating, fetal, and neonatal livers. These changes are discussed in terms of gene expression relevant to malignant transformation of hepatic cells.     

Early Symptoms of Autistic Children and Its Diagnostic Significance

Abstract. In order to make a checklist for the early diagnosis of autism, the abnormalities of mental development before the age of two were examined retrospectively in autistic children. Additionally, the correlation between these early symptoms and the prognosis of autistic children were investigated. The subjects were 85 autistic children, 64 mentally retarded children and 150 normal children. The parents of these children were interviewed by child psychiatrists based on original questionnaires. The 27 early symptoms indicating disorders in personal relations, perception and sleeping as well as the setback course were found in autistic children. The onset ages of imitating others, pointing at objects and saying Yes were retarded in the poorly developed group as compared with the highly developed group.