Molecular analysis of digenic inheritance in Bartter syndrome with sensorineural deafness.

BACKGROUND: Bartter syndrome (BS) is a genetic disorder accompanied by hypokalaemic metabolic alkalosis. BS with sensorineural deafness (SND, OMIM602522) is a newly identified phenotype caused by mutations in the BSND gene that encodes barttin, a beta-subunit for chloride channel ClC-Ka and ClC-Kb and classified as type IV BS. Type IV BS features the most severe phenotype entailing life-threatening neonatal volume depletion and chronic renal failure developing during infancy. A recent report described a case of BS with SND from a consanguineous family who showed homozygous mutations in the CLCNKA and CLCNKB genes. This case indicated the possibility of the occurrence of digenic inheritance in BS with SND resulting from double mutations in the CLCNKA and CLCNKB genes. SUBJECT AND RESULTS: The current report concerns a 2-year-old girl from a non-consanguineous family with BS accompanied by SND. In our case, four loss-of-function mutations, consisting of mutations in both parental alleles in both CLCNKA and CLCNKB, were identified. The paternal allele had a nonsense mutation (Q260X) in CLCNKA and a splicing site mutation (IVS17+1 g>a) in CLCNKB. The maternal allele had a large deletion mutation (about 12 kbp) extending from CLCNKA to CLCNKB. Our case provides clear evidence that loss-of-function alleles in both alleles of both CLCNKA and CLCNKB results in a phenotype indistinguishable from that of mutations in BSND (type IV BS). CONCLUSIONS: Recent advances in genetics have resulted in a better understanding of many human inherited diseases, but most of them are monogenic disorders and more complex inheritance patterns remain unresolved. Our case provides clear evidence of digenic inheritance outside the scope of Mendelian inheritance disorders.

Adriamycin-lipiodol suspension for i.a. chemotherapy of hepatocellular carcinoma.

Physicochemical properties of two types of adriamycin preparation, suspensions and emulsions prepared for i.a. chemotherapy of hepatocellular carcinoma, were investigated. A suspension was prepared by dispersing adriamycin directly into the lipid contrast medium, Lipiodol, whereas an emulsion was obtained by emulsifying an aqueous solution of adriamycin into Lipiodol. The dispersibility of the drug in each preparation was examined microscopically. The chemical stability of and drug release from the preparation were determined by high-performance liquid chromatography and spectrophotometry, respectively. The suspension was then given to ten patients with primary hepatocellular carcinoma. The suspension maintained good dispersibility without coagulation of drug particles, whereas coalescence of aqueous droplets and the resultant phase separation occurred 4 h after preparation of the emulsion. Both preparations maintained the initial drug content for at least 1 week at room temperature. The release of adriamycin was more prolonged in the suspension than in the emulsion. After i.a. administration of the suspension, a selective accumulation of Lipiodol in the tumor and decrease in serum alpha-fetoprotein (AFP) levels were found in most patients. A significant amount of adriamycin was still detected in hepatic specimens resected from two patients 1 and 2 months after treatment. These findings suggest that the adriamycin-Lipiodol suspension may be a useful preparation for targeting chemotherapy to hepatocellular carcinoma.

Biological, serological, and molecular variabilities of clover yellow vein virus.

ABSTRACT A comparative study was made on the host reactions, serological properties, and nucleotide sequences of the coat protein (CP) gene of 10 clover yellow vein virus (C1YVV) isolates and one bean yellow mosaic virus (BYMV) isolate collected from different host plant species and locations in Japan. Two strains of C1YVV isolates, grouped on the basis of host reactions on Chenopodium amaranticolor, C. quinoa, Nicotianaclevelandii, N. benthamiana, Vicia faba, and Trifolium repens, corresponded to two serotypes determined by double-antibody sandwich- and triple-antibody sandwich-enzyme-linked immunosorbent assay using three polyclonal and nine monoclonal antibodies. These results were also confirmed by nucleotide sequence analysis of the CP gene. The CP gene of C1YVV isolates of strain 1, including the Australian isolate C1YVV-B, had 93 to 98% nucleotide identities and 97 to 99.6% amino acid identities. The CP of C1YVV isolates of strain 2, including the New Zealand isolate C1YVV-NZ, had 92 to 98% nucleotide identities and 95 to 98% amino acid identities. The nucleotide identities and the amino acid identities between the two C1YVV strains were 82 to 84%, and 90 to 94%, respectively. When compared with the CP sequences of 12 C1YVV isolates, the CP sequence of the BYMV isolate had 71 to 73% nucleotide identity and 73 to 77% amino acid identity. Amino acid sequence differences among C1YVV isolates from strains 1 and 2 were located mostly at the N-terminal regions of the CP. Our results indicated that the C1YVV isolates studied could be separated into two strains on the basis of host reactions, serology, and the nucleotide sequence of the CP gene.

[Epidemiological study on respiratory effects of smoking in college students].

To study the influence of smoking on the respiratory system in adolescents, we conducted a prospective study to look into the respiratory symptoms, changes in ventilatory function, and levels of expired CO of both smoking and nonsmoking college students. The subjects were 56 male students who entered Akita University in 1986. Thirty of the participants were smokers at entrance whereas the rest who were nonsmokers, served as controls. The smokers started smoking at an average age of 17.6 +/- 0.9 years. Physical examinations at 8 points during the study were performed periodically in May and November for four years, from 1986 to 1989. From these studies, the following conclusions were obtained. 1) The prevalence of respiratory symptoms, especially phlegm, was higher among the smokers compared with the nonsmokers; the differences not being significant. Among both the smokers and nonsmokers, no prevalence of persistent cough and phlegm was observed during the study period. 2) There were no differences in the levels of %FVC and %V50 between the smokers and the nonsmokers at any point during the study period. The FEV1% levels for smokers were decreased progressively, and the differences became significant at the 7th and 8th tests (p < 0.05). After the second test, %V25 levels were lower in the smokers than in the nonsmokers; the differences being significant at the 7th test (p < 0.05). 3) The dose-response relationships between smoking and pulmonary function were not statistically significant in any levels at the 7th and 8th tests. However, %V25 levels of the heavy smokers (15 < or = per a day) were lower than the those of the nonsmokers; the differences being significant at the 7th and 8th tests (p < 0.05). 4) Average expired CO concentrations of the smokers were 2.5-3.5 times higher than those of the nonsmokers during the study period, confirming exposure to tobacco smoke. These observations suggest that young smokers may develop obstructive changes, especially narrowed small airways during the first 5 years of exposure to tobacco smoke.

[A review of studies on school-based smoking prevention programs].

Research papers on school-based programs designed to prevent adolescent smoking were reviewed to determine trends in the development of smoking prevention programs as well as measures and methods of evaluation. In the USA and Europe, evaluation studies have demonstrated that the most promising smoking prevention approaches are those that focus primary attention on the psychosocial factors promoting smoking initiation. In Japan, however, most programs have shown little success. The following recommendations are made for improving the methods for evaluating program effectiveness and the future of research orientation of educational program on adolescent smoking. 1) Refinement and standardization of evaluation methods to be valid and feasible for use in studies of Japanese adolescents. 2) Improved study designs for evaluation of program effectiveness which include control groups and long term follow-up. 3) Applying smoking prevention programs which were demonstrated to be effective in the USA and Europe. 4) Development of effective cessation programs for adolescent smokers.

[Knowledges of foodstuffs in relation to adult diseases in Japanese students--results from Japan Know Your Body Study].

As part of developing the Know Your Body (KYB) program for Japan, a questionnaire survey was administered assessing knowledge, attitudes, and behavior regarding cigarette smoking, alcohol drinking, dietary lifestyle and exercise. The subjects, about 13,000 in total, were students in elementary, junior and senior high schools in 9 prefectures. The main results concerning knowledge of foods that cause or prevent adult diseases are as follows: 1) Correct responses on the relationship of excessive salt intake and hypertension were made by 39% of males and 41% of female in the fifth grade of elementary school. In the third grade of junior high school, the percentage of correct answer for the same question increased to 80% of males and females, and in the third grade of senior high school, it rose to 90% of males and 91% of females. No clear sex difference was seen in all the grades of school. 2) Less than 23% of the students from the first to fourth grade of elementary school knew the word "cholesterol" and could distinguish high cholesterol foods. In the sixth grade of elementary school, more than 50% of students knew the word "cholesterol". In the third grade of junior high school, more than 50% of students could distinguish high cholesterol foods correctly and recognized the relationship of excessive cholesterol intake to heart disease. These results provide a basis for determination of the age for introducing nutrition education. 3) The percentage of correct answers for distinguishing green-yellow vegetables, showed a clear sex differential. The percentage of correct answers concerning dietary fiber was 75% for males and 81% for females in senior high school.

[Standardization of definitions concerning smoking behavior among Japanese adolescents--results from the Japan Adolescent Smoking Survey (JASS)].

The aim of the Japan Adolescent Smoking Survey (JASS) is to standardize the definitions of smoking behavior among Japanese adolescents together with measurement methods and questionnaires concerned. A study of 9921 school children aged 9 to 18 years in 17 elementary schools, 16 junior high schools and 18 senior high schools in 9 prefectures, was conducted between June and July of 1989, to determine the relationship between actual smoking behavior during the past month or year and the "self-referent label" concerning their smoking status. The main results were as follows: 1) The percentage of those who smoked at least one cigarette during the past month showed a high degree of similarity to the percentage of those who referred to themselves as "regular smokers" or "occasional smokers". 2) Less than 10% of students who identified themselves as "never smokers" or "ex-smokers" smoked at least one cigarette during the past month, while more than 80% of students who referred to themselves as "regular smokers" or "occasional smokers" smoked during the past month. 3) On the whole, those who smoked 20 or more cigarettes during the past month referred to themselves as "regular smokers"; those who smoked between 2 and 19 cigarettes as "occasional smokers"; those who smoked 1 cigarette as "ex-smokers"; those who did not smoke during the past month as "never smokers". 4) The definition where students who smoked one cigarette during the past month are classified as a category of smoker, displayed a high level of sensitivity and specificity for all school-level groups and for both sexes.(ABSTRACT TRUNCATED AT 250 WORDS)