Changing patterns of cancer and diet in Japan.
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Biomedical subjects
Publications and source records attributed to I Kato.
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A simple and reliable method is described for direct sequencing of material generated by the polymerase chain reaction. The protocol is based on the purification of the amplified double-stranded product by polyethylene glycol precipitation, annealing of primer with template by a "snap-cooling" procedure and sequencing by the dideoxy chain termination method with the use of Klenow fragment or Taq polymerase. The limit of the size of PCR products that can be sequenced is also discussed.
Recently, the two Mr forms of caldesmon (Mr's in the range of 120-150kDa and 70-80kDa as judged by SDS-PAGE) have been identified. h-Caldesman (high Mr 120-150kDa caldesmon) is predominantly expressed in smooth muscles, and l-caldesmon (low Mr 70-80kDa caldesmon) in non-muscle cells. In this paper, we report the nucleotide sequence of chick embryo gizzard h-caldesmon cDNA and its translation into amino acid sequence. This sequence predicts a protein of 771 amino acids with a Mr of 88,743. The central portion of this sequence is composed of a 10-fold repeat of conserved amino acid sequence containing 13-15 amino acids. Further, a recombinant protein produced in Escherichia coli containing the full-length h-caldesmon cDNA has been characterized. Although the Mr of h-caldesmon predicted from amino acid sequence is 88,743, native and recombinant proteins show the same mol. wt. with 150kDa as measured by SDS-PAGE. This discrepancy may be due to the acidic amino acid-rich sequences at the N-terminal and central portions. A recombinant protein produced in E. coli possesses calmodulin-, F-actin- and tropomyosin-binding abilities in common with the native h-caldesmon.
A tetrapeptide Arg-Gly-Asp-Ser (RGDS) has been shown to be a versatile cell recognition signal of extracellular matrix components for the interaction with cells. We introduced the RGDS tetrapeptide into a truncated form of protein A, a staphylococcal immunoglobulin-binding protein, by inserting an oligonucleotide cassette encoding the tetrapeptide into the coding region of the protein A expression vector pRIT2T. The mutagenized protein was capable of not only binding to immunoglobulin G but also mediating cell attachment and spreading onto an inert substrate. Cell adhesion mediated by the mutagenized protein was inhibitable by a synthetic peptide Gly-Arg-Gly-Asp-Ser but not by a related peptide Gly-Arg-Gly-Glu-Ser, confirming that the inserted RGDS tetrapeptide served as a recognition signal for cell adhesion. Furthermore, the RGDS-containing protein was capable of adhering cells onto an immunoglobulin-coated surface which could not by itself support cell adhesion. Thus, the cell adhesive and immunoglobulin binding activities of the mutagenized protein appear to function coordinately. The protocol described here is essentially applicable to any protein and, therefore, provides a general principle in tailoring novel multifunctional proteins having cell adhesive activity.
Staphylococcal alpha-toxin resulted in ADP-ribosylation of the 37 and 41 kDa proteins of a membrane preparation from rabbit erythrocytes. In the presence of 100 microM GTP, the toxin ADP-ribosylated proteins of 54 and 59 kDa and potentiated ADP-ribosylation of the 37 and 41 kDa forms. GTP had no effect on ADP-ribosylation of membrane proteins in the absence of alpha-toxin. Incubation of a membrane preparation of rabbit polymorphonuclear leukocytes with the S and F components of staphylococcal leukocidin resulted in ADP-ribosylation of the 37 and 41 kDa proteins, respectively. Furthermore, the 37, 41, 54 and 59 kDa proteins were ADP-ribosylated by leukocidin in the presence of GTP. The ADP-ribosylation of these proteins was observed to be dependent on the incubation time and toxin dose and was abolished by prior boiling. Addition of agmatine did not attenuate ADP-ribosylation of these proteins. These results demonstrate that staphylococcal alpha-toxin and leukocidin possess ADP-ribosyltransferase activities which are potentiated by GTP and suggest that ADP-ribosylation reactions are responsible for development of the cytolytic activities of these staphylococcal toxins.
Using a tropomyosin-coupled affinity column, we have demonstrated a direct association between the chymotryptic 35 kDa fragment of h-caldesmon, which is located at the C-terminal of the parent molecule, and gizzard tropomyosin. We have subsequently determined the nucleotide sequence of cDNA clones encoding the 35 kDa fragment from the cDNA library prepared from chick embryo gizzards, and have deduced the amino acid sequence. Calculating from the predicted sequence, the 35 kDa fragment is composed of 306 amino acid residues. In agreement with the tropomyosin-binding ability, the 35 kDa fragment conserves two consensus sequences of the tropomyosin-binding domain in troponin T. These results suggest that the 35 kDa fragment of h-caldesmon, at least in part, has a common property to the striated muscle troponin T.
Two monoclonal anti-fibronectin antibodies that inhibit fibronectin-mediated cell adhesion have been established and characterized. One antibody, FN12-8, inhibited attachment of rat kidney fibroblasts on the fibronectin-coated substrate in a concentration-dependent manner, attaining a maximal inhibition of greater than 85% at 850 micrograms/ml. Another antibody, FN30-8, caused about 70% inhibition at a concentration as low as 0.85 microgram/ml, although further increase of the antibody concentration did not significantly augment the inhibitory effect. Immunoblot analysis with defined proteolytic fragments revealed that both antibodies are directed to the cell-binding domain of fibronectin. The epitopes for these antibodies were further narrowed down using recombinant cell-binding fragments expressed in Escherichia coli. FN12-8 recognized the 11.5-kDa cell-binding fragment previously characterized by Pierschbacher et al. (1981, Cell 26, 259-267), suggesting that FN12-8 blocks the Arg-Gly-Asp (RGD) cell adhesion signal. FN30-8 could not bind this fragment but did recognize a longer cell-binding fragment containing additional greater than 111 amino acid residues upstream of the 11.5-kDa fragment. Since the RGD-dependent cell adhesion seems to require another signal located at a region 50-160 residues upstream of the 11.5-kDa fragment for full activity, FN30-8 may exert its inhibitory effect by blocking the latter signal.
Three examples of malignant neoplasms primary to the oral cavity and associated with paraneoplastic syndromes are presented. The first case is a squamous cell carcinoma of the maxilla associated with leukocytosis. The second case is a mandibular squamous cell carcinoma associated with hypercalcemia in the absence of bony metastases. The third case is a squamous cancer of the tongue that metastasized to the lumbar vertebrae and right second rib and was associated with both hypercalcemia and leukocytosis. There was no evidence of acute infection or leukemia that could be expected to account for leukocytosis. Hypercalcemia in the second case was defined as humoral hypercalcemia of malignancy by biochemical and clinical evaluations. To our knowledge, this is the first definitive report of a carcinoma primary to the oral cavity associated with humoral hypercalcemia of malignancy. In each case, the severity of hypercalcemia, leukocytosis, or both very closely correlated with tumor growth. Surgical excision of the tumors or regression of tumor mass due to aggressive anticancer drug administration resulted in decreases in leukocyte number, serum calcium level, or both. In contrast, recurrence or regrowth of tumors induced further development of hypercalcemia, leukocytosis, or both. It is therefore likely that humoral factors released by these oral carcinomas are responsible for the hypercalcemia, leukocytosis, or both.
Major categorical diagnosis by International Classification of Diseases and type-specific diagnosis for cardiovascular diseases in death certificates were compared to the diagnosis made at autopsy in 864 consecutive autopsy cases aged 20 or over, among the Japanese residents in Hisayama town. Cerebral stroke was correctly diagnosed in 84%, malignant neoplasms in 78% and cardiac disease in 66%. Cerebral stroke and cardiac disease tended to be overdiagnosed, while malignant neoplasms were underdiagnosed. The validation of certified diagnosis was less reliable in the aged population, and in type-specific diagnosis of cardiovascular diseases. Cerebral hemorrhage with false negative or false positive diagnoses was usually classified into type unspecified stroke or different categories of cerebral stroke, while those misdiagnosed as cases of cerebral infarction frequently had no significant lesions in the autopsied brain. Finally, the relationship between the validation of diagnosis on the death certificates and the secular trend in cardiovascular disease in the Japanese vital statistics was discussed.
Milia developed in two patients with different chronic dermatoses after 2 weeks of treatment with clobetasol propionate ointment. Routine histologic and immunohistochemical analysis showed the cysts to be of eccrine gland origin.
Chicken, human(alpha) CGRPs and their analogues were synthesized to investigate the relationship between structure and serum calcium and phosphate lowering effects. The native hormones contain 37 amino acid residues with only four positions (3, 14, 15 and 23) that differ. Chicken CGRP exhibits stronger and longer lasting activities than the human hormone. A chicken CGRP analogue with replacement of Asp at position 14 by Gly (as in human CGRP) showed great reduction of activity. The converse replacement of Gly at position 14 by Asp in human CGRP enhanced this analogue activity. Among the synthetic analogues, des-1-Ala-des-alpha-amino chicken CGRP, exhibited the most potent and long lasting biological activity.
We studied characteristics of past smokers according to the duration of cessation of smoking based on data from a population-based survey. Lifestyle, prevalences of various symptoms and diseases and other factors were compared among current smokers (8507 males and 2012 females), past smokers (4423 males and 684 females) and non-smokers (2431 males and 12,859 females) aged 40 years and over. Compared to current smokers, past smokers consumed more bread, milk, vegetables, fruit and black tea, and less rice, pickles, instant noodles, coffee and alcohol, had lower prevalences of cough, sputum and anorexia, participated more in cancer screening tests, weighed more, included more professional and administrative workers and had more non-smoking spouses. These characteristics resembled those of non-smokers. But past smokers had high prevalences of several cardiovascular and respiratory diseases compared to current smokers. Daily intake of coffee was inversely associated and daily intakes of fruit and milk were positively associated with the duration of abstinence from smoking after adjusting for other factors in both sexes. These results suggest that lifestyle of past smokers may contribute to risk reduction for several diseases.
Two overlapping cloned cDNAs encoding the entire amino acid sequences of the subunits of acylamino acid-releasing enzyme (AARE) [EC 3.4.19.1] have been isolated from porcine liver cDNA lambda gt10 cDNA libraries and sequenced. Sequence analyses of the cDNA and several Achromobacter protease I-digested peptides of the purified protein revealed that porcine liver AARE consists of four identical subunits, and each comprising a single chain of 732 amino acids with acetylmethionine at the N-terminus.
A case-control study of biliary tract cancer was conducted in Niigata prefecture where the mortality of the cancer is the highest in Japan. The cases were 109 patients with gallbladder cancer and 84 with bile duct cancer, and the controls were 386 sex- and age-matched neighborhood controls. For gallbladder cancer, a past history of biliary tract disease, a positive family history of cholelithiasis and a taste for oily foods were high risk factors. Intakes of animal proteins and fats such as fish, eggs, meat, etc., ingestion of vegetables and fruits, and taking snacks were low risk factors for gallbladder cancer. For bile duct cancer, a past history of biliary tract disease, a family history of cerebral vascular accident, a thin constitution and taking a small amount of foods were high risk factors, and a family history of heart disease, obesity, intakes of alcohol, animal proteins and fats, or frequent intakes of vegetables and fruits were low risk factors.
The relationship between marital status and cancer incidence was examined based on 49,191 incident cases aged 30 or over in 1980-1984 by using the data from Aichi Cancer Registry and census data. Although married and widowed people did not show increased incidence for any cancer site studied, single and divorced people showed statistically significantly increased or decreased risks for several sites of cancer. Single males showed an increased risk for esophageal cancer and a decreased risk for lung cancer. Divorced males showed increased risks for cancers of the mouth & pharynx, esophagus, liver, skin and brain. Single females showed increased risks for cancers of the esophagus, stomach, small intestine, liver, pancreas, lung, breast, corpus uteri, ovary & fallopian tube and other female genital organs and a decreased risk for cervical cancer. Divorced females showed increased risks for cancers of the larynx, breast, all parts of uterus and cervix uteri and a decreased risk for biliary tract cancer. The increased risk for breast cancer in single females was more pronounced in older age groups and the increased risks for several sites of cancer in divorced people were more pronounced in younger age groups. These findings may be partly explained by differences in reproductive factors and life style, especially smoking and drinking habits.
A hair matrix tumor showing an unusual tumor cell arrangement was found at the base of a solitary trichoepithelioma. Coexisting with solid epithelial islands and immature hair follicle-like stroma resembling the Verocay bodies of neurilemmoma or "ripplemarks" on waves were found. In other areas myxomatous degeneration of the stroma changed the rippling into a cribriform pattern. In some parts of the tumor there was a dense melanin pigment associated with MEL5 stained melanocytes. S-100 and CD1 (OKT6) antigen stains demonstrated Langerhans cells scattered in the parenchyma and less frequently in the stroma. The majority of tumor cells were considered immature pilar cortical cells because of the following: 1. HKN-6 was strongly positive; 2. a large number of melanocytes were associated with tumor cells in some foci; 3. ultrastructurally immature tumor cells, which had electron-dense tonofilaments and many desmosomes, were transformed without production of trichohyalin granules into semikeratinized cells which showed nuclear degeneration and loss of electron density in tonofilaments. This tumor, however, has not attained the degree of differentiation observed in trichoblastoma (1) another example of an immature cortical cell tumor. Squamous eddy-like or horn pearl-like foci of incomplete keratinization and large keratin-filled cysts were also present within the immature parenchyma, indicating that some immature cells were differentiating toward non-cortical cells, as found in the outer sheath. We would like to designate this tumor "rippled pattern trichomatricoma", a new entity.
Several monoclonal antibodies (MAB) have been produced using an eccrine carcinoma cell line as an immunogen. One such MAB, SKH1, reacted with both the secretory portion and coiled duct of the eccrine and with the secretory portion of apocrine gland. SKH1, however, did not react with myoepithelial cells, intradermal ducts of both types of sweat gland, or with other components of normal axillary skin including the epidermis and follicular apparatus. The reaction was strongest if the specimen was fixed with 80% methanol, and moderate on non-fixed or acid-alcohol-fixed specimens. Only weak reaction was obtained on cold acetone-fixed specimens, and reaction was negative with formalin-fixed, paraffin-embedded tissues. SKH1 reacted positively with the cytoskeleton of the eccrine carcinoma cell line, Colo-16 and MCF-7. Applied to pathological skin specimens, SKH1 reacted with the tumor cells of clear cell hidradenoma, syringocystadenoma papilliferum, and extramammary Paget's disease. SKH1 also reacted with the tumor cells of metastatic adenocarcinomas arising from lung, breast and ovary. SKH1 did not react with the majority of tumor cells of eccrine poroma, but reacted with single-layered cells lining narrow ductal lumina. SKH1 did not react with epithelial cells lining cystic or ductal lumina of syringoma, but reacted moderately with the amorphous keratin-like substance filling the lumina. Immunoblot analysis revealed that SKH1 recognizes a 40 Kd sweat gland-associated antigen, and can be an aid to identifying tumors arising from sweat gland structures.
Twenty-six first episodes of subarachnoid hemorrhage occurred among 1,621 Hisayama residents aged greater than or equal to 40 years during the 22-year follow-up of a prospective study. Subarachnoid hemorrhage was confirmed by both clinical and autopsy findings. The average annual incidence (96.1/100,000 population) was 3-13 times higher than any previously reported and steeply increased with age in both sexes, being 2.3 times higher for women than for men after adjusting for age. Nine patients (35%) died less than or equal to 8 hours after the onset of subarachnoid hemorrhage. None was correctly diagnosed on the death certificates, and four of the nine (44%) were misdiagnosed as intracerebral hemorrhage. We found the survival rate of patients suffering subarachnoid hemorrhage to be much lower than previously reported because we detected a large number of sudden deaths due to subarachnoid hemorrhage through the high rate of autopsy in our cohort (81.4%).