[Bronchial asthma related to the mechanism of allergy--1) Genetic immunology of atopic factors--abnormality of the regulation of IGE antibody production].
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Biomedical subjects
Publications and source records attributed to K Joh.
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The complete nucleotide sequence of the rat aldolase A isozyme gene, including the 5' and 3' flanking sequences, was determined. The gene comprises ten exons, spans 4827 base-pairs and occurs in a single copy per haploid rat genome. The genomic DNA sequence was compared with those of three species of rat aldolase A mRNA (mRNAs I, II and III) that have been found to differ from each other only in the 5' non-coding region and to be expressed tissue-specifically. It revealed that the first exon (exon M1) encodes the 5' non-coding sequence of mRNA I, while the second exon (exon AH1) encodes those of mRNAs II and III and the following eight exons (exons 2 to 9) are shared commonly by all the mRNA species. These results allowed us to conclude that mRNA I and mRNAs II, III were generated from a single aldolase A gene by alternative usage of exon M1 or exon AH1 in addition to exons 2 to 9. S1 nuclease mapping of the 5' ends of their precursor RNAs suggested that these three mRNA species were transcribed from three different initiation sites on the single gene.
Three species of aldolase A mRNA (mRNAs I, II, and III) only differing in the structure of the 5'-terminal noncoding region were detected in rat tissues. The cDNA clones for mRNAs II and III were prepared from ascites hepatoma AH60C and sequenced. The mRNA II is 1393 nucleotides long excluding poly(A) tail, while the mRNA III is 1440 nucleotides long, some 50 nucleotides longer than the mRNA II. The mRNAs II and III differ in the sequence between -25 and the 5' termini from the previously reported skeletal muscle aldolase A mRNA (mRNA I, 1343 nucleotides long). By contrast, the residual 5' noncoding sequence (-24 to -1) and the coding and 3' noncoding sequences are common to all the mRNAs. By dot spot hybridization and S1 mapping the distribution of these mRNAs in the various tissues was determined. The mRNA I appears exclusively in a skeletal muscle and some in heart and hepatoma AH60C, whereas the mRNAs II and III appear more or less in all the tissues examined, implying that their appearances are under tissue-specific control. Furthermore, partial nucleotide sequence analysis of the fetal liver aldolase A mRNA supports that aldolase A mRNA that reappeared in hepatoma is really a resurgence of the gene product expressed in the fetus.
The Nuclear Body Appearance Rate in seventeen human prostatic cases was statistically analyzed according to three lesion areas--the hyperplastic nodule, non-nodule and atrophic--in the secretory epithelium and basal cells. It was meaningfully high in the secretory epithelium of the hyperplastic nodule, but not in the other two lesion areas. There was no meaningful result in basal cells. Though there is a wide variety of reported data on the human prostate, particular care must be taken in analysis at the electron microscopic level, keeping in mind that even in a single specimen this organ has a notable variety of tissue changes. The Nuclear Body Appearance Rate reflects cellular hyperactivity although it does not have a specific known function at present.
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Fundamental and clinical studies were carried out on cefixime (CFIX) 5% granules, and the results are summarized below. Antimicrobial activity Antimicrobial activities of CFIX, cefaclor, cefroxadine, cephalexin and amoxicillin (AMPC) were studied against clinical isolates. CFIX showed greater activities than all the other antibiotics against E. coli, K. pneumoniae, H. influenzae, P. mirabilis, E. cloacae and S. marcescens, but it was slightly less active than AMPC against S. pyogenes. Absorption and excretion Serum concentrations and urinary excretions of CFIX were determined following single or repeated oral administration. In 8 patients given single dose of CFIX 1.5 or 3.0 mg/kg, mean serum concentrations were 1.27 and 1.09 micrograms/ml at 2 hours, 1.27 and 1.35 micrograms/ml at 4 hours, 0.85 and 1.10 micrograms/ml at 6 hours, 0.17 and 0.24 micrograms/ml 12 hours after administration, respectively. Mean serum half-lives were 2.54 hours for the dose of 1.5 mg/kg and 2.60 hours for 3.0 mg/kg. Urinary recovery rates in the 12-hours urine varied 6.7 to 33.6%, with an average of 13.5%. In 3 patients given a repeated dose of CFIX 3.0 or 5.6 mg/kg b.i.d., the serum concentrations were 0.23-1.01 micrograms/ml at 0 hour, 1.91-2.80 micrograms/ml at 2-4 hours and 1.13-2.07 micrograms/ml at 6-8 hours after administration. Clinical study The CFIX was given orally by mainly b.i.d. at a daily dose of 4.4-11.6 mg/kg for 4-15 days to a total of 33 patients consisting of 3 patients with pneumonia, 3 with bronchitis, 9 with tonsillitis, 15 with UTI, one each with scarlet fever, lymphadenitis and colitis. Clinical responses were excellent in 24 patients, good in 8 and fair in 1, with an effectiveness rate of 97.0%. All of the 21 bacterial isolates examined were eradicated after CFIX treatments including 3 beta-lactamase producing strains. No side effects of abnormal laboratory findings were observed in these patients.
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The nucleotide sequence of aldolase A mRNA in rat skeletal muscle was determined using recombinant cDNA clones and a cDNA synthesized by primer extension. The sequence is composed of 1343 nucleotides (nt) except for the poly(A) tail. Based on the sequence analysis we have deduced an open reading frame with 363 amino acids (aa) (Mr 39134). The sequence suggests several nt polymorphisms in the mRNA population, one of which causes an aa change. The determined sequence of rat aldolase A mRNA was compared with the published ones of rabbit aldolase A or rat aldolase B mRNAs. The homology between rat and rabbit aldolase A mRNA sequences is greater than that between rat aldolase A and B mRNA sequences. Multiple aldolase A mRNAs having different Mrs were detected in the various tissues, and appeared to be expressed in a tissue-specific manner. Further analysis suggests that differences in mRNA length are due to differences in the 5'-noncoding terminal region.
A muscle-type aldolase gene known to be a tumor marker enzyme was analyzed. It was found that three different molecular species of the mRNA exist and are expressed in a tissue-specific manner. These mRNAs have identical coding and 3' noncoding sequences and differ only at the 5' end of the sequence. Genomic DNA analysis indicated that a single aldolase gene for one muscle type specifies three different mRNAs by organizing as a leader sequence a region corresponding to each distinct 5' end of the mRNA followed by a shared common structural gene in the genome.
Rat aldolase A mRNAs from the skeletal muscle and ascites hepatoma cells are compared by RNA blot hybridization. The hepatoma AH60C and AH130 mRNA are about 1600 bases long, some 50 bases longer than the skeletal muscle transcript. Aldolase A enzymes encoded by these mRNAs, however, are indistinguishable in electrophoresis on cellulose polyacetate strips and two-dimensional polyacrylamide gel. In addition, comparison of the thermal stability of hybrids between the mRNAs from the skeletal muscle and hepatoma cells and a cDNA probe (pRAAM83) made from the aldolase A mRNA of the skeletal muscle indicate that the mRNAs have extremely high sequence homology, at least, within the most part of coding region in addition to the 3' noncoding region. These findings indicate that these mRNA may be transcribed by the use of different promoter sites and the common structural gene.
Endotoxin induced necrosis of the Meth A mouse tumour has been investigated using macroscopic, histological and ultrastructural examination methods. On the 8th day after tumour cell transplantation, the animals received a relatively non-toxic dose of the Salmonella abortus equi endotoxin intravenously. The natural history of the tumour necrosis took the following course: The earliest morphological changes could be seen with the electron microscope 90 min after administration of the endotoxin, and were seen as an interstitial oedema with separation of the tumour cells. Haemorrhagic necrosis of the tumour was complete 4 hours after injection, and could be easily recognized with the naked eye. Rejection of the necrotic malignant tumour was complete two weeks after LPS administration. Only minor residual scarring of the belly-wall remained. Haemorrhagic tumour necrosis due to endotoxin can be compared with the localized Shwartzman reaction and probably involves tumour necrotizing factor (TNF). For complete destruction of a tumour by haemorrhagic necrosis the size of the tumour is critical. Certain regression after endotoxin administration depends upon additional T-cell-mediated immunity (provided the tumour is immunogenic). In contrast to the haemorrhagic necrosis, BCG-induced tumour regression is accompanied by granulomatous inflammation, which may be responsible for destruction of the tumour.
Two- and three-dimensional electron microscopic observations by a serial sectioning method revealed endothelial sprouts with intracytoplasmic vacuolization in rapidly growing human juvenile hemangioma. A large vacuole bounded by a single unit membrane was enclosed in the cytoplasm and on the inner aspect of the vacuolar membrane several short microvilli were demonstrated. These appearances have not been reported before. The presence of microvilli in the vacuole indicates that the endothelium has reached the point of differentiation when a vascular lumen forms. In the cytoplasm adjacent to the vacuolar membrane, a significant number of 7 to 10 nm microfilaments were identified. These intracytoplasmic microfilaments are assumed to play a mechanical role in the development of the cytoplasmic vacuole and/or the sprout. The formation of a vacuole observed in the endothelial sprout is similar to the findings of Sabin (1920) by light microscopy in endothelial sprouts in the blood island of the chick embryo. The active endothelial sprout in juvenile hemangiomas is considered to be at least partially responsible for the capillary proliferation and enlargement of the tumor.
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The Niemann-Pick mice which received bone marrow transplants showed a decreased accumulation of sphingomyelin and cholesterol quantitatively in their spleen. The sphingomyelin deposit in the bone marrow was also reduced histochemically. However, the neurological manifestations were not improved by the bone marrow graft.
Local or systemic prostaglandin (PG) administration leads to the known softening and dilatation of the cervix uteri. Lysosomal enzymes are involved in connective tissue degradation. The question arises whether the effect of PG on the cervix uteri is mediated by lysosomes. Five pregnant women (volunteers after informed consent) in the first trimester received 500 micrograms of PGE2-derivative (Nalador) i.m. at 12 and 8 h before termination by curettage. Five pregnant women without PG-treatment served as controls. Small biopsies were obtained from the endocervical canal and were immediately immersed in cold 2.5% glutaraldehyde and after further preparations examined under a Zeiss electron microscope 9S-2. A second portion of tissue was sliced and prepared for histochemical analysis of the acid phosphatase on lysosomes. Examination of the ultrastructure of the cervix uteri showed vesicles in the extracellular matrix. These were surrounded by a single membrane and contained either fine granular material of myelin-like whorls of membranes. These vesicles lay between collagen fibers, showed the reaction product of acid phosphatase and were often surrounded by an electron-lucent halo. We conclude that these matrix vesicles were "matrix lysosomes" extruded from the cervical myo-fibrocytes into the extracellular space as a result of the PG-E2-administration. Here they are not under cellular control and can initiate the proteolytic degradation of connective tissue. This might be the crucial step in cervical dilatation which, on ultrastructural examination, can be seen as decreasing electron density of the extracellular ground substance near the matrix lysosomes. The relationship between PGE2 and collagenase production is generally accepted. If one believes that lysosomal cathepsin D and cathepsin B act synergistically with collagenase, it can be assumed that PGE2 is involved in a lysosomal degradation of the connective tissue. The morphological sign of this occurrence is the release of matrix lysosomes by PGE2 as described in the present study. Extracellular lysosomes and their physiological significance in cervical function are discussed in detail.
A case of a 66 years old female who had a nephrogenic adenoma recurring three times during a 6 months course of her illness is reported. With each recurrence, the lesion moved toward the urethral orifice, suggesting an intimate relationship between the recurrent process and physical stimuli to the mucosa. Electron microscopy revealed merely non-specific morphological features of epithelial cells in the lesion. There were no positive morphological clues in the lesion for supporting the lesion to resemble mesonephric and/or mesodermal tissue. The diagnostic term of tubular metaplasia for the lesion is preferable instead of nephrogenic adenoma.