Masaru Miki

Genetic differences in CYP2C19 single nucleotide polymorphisms among four Asian populations

Background. This study was designed to compare genetic differences in single-nucleotide polymorphisms of the S-mephenytoin 4′-hydroxylation (CYP2C19) gene among four Asian populations. Methods. Polymerase chain reaction with restriction fragment length polymorphism (PCR-RFLP) analysis of CYP2C19 was conducted in Japanese, Chinese, Thai, and Vietnamese populations. All genotype frequencies were analyzed. Wild-type homozygote and wild-type heterozygote genotypes were extensive proton pump inhibitor (PPI) metabolizers. Mutant-type heterozygote and mutant-type homozygote genotypes were poor PPI metabolizers. Results. No significant differences in CYP2C19 phenotype, calculated based on genotype frequencies, (P > 0.05) were found among the four populations. Conclusions. Many factors, including CYP2C19 polymorphisms, affect the success rate of Helicobacter pylori eradication with PPI-based therapy. We suspect that CYP2C19 polymorphisms may not be the main factor associated with differences among these four Asian populations in the success rates of H. pylori eradication with PPI-based therapy.

Gastrospirillum hominis and Helicobacter pylori infection in Thai individuals: Comparison of histopathological changes of gastric mucosa

The presence of Helicobacter pylori (H. pylori) in the stomach is closely associated with histological signs of chronic active gastritis and peptic ulcer. Another spiral organism named Gastrospirillum hominis (G. hominis) has led to further interest in the bacterial pathogenesis of gastritis. Due to the low prevalence of G. hominis, it is difficult to evaluate its biological behavior. Recently 16 cases of G. hominis‐associated gastritis were found in 257 Thai individuals, which made it possible to study the biological characteristics of G. hominis and its relationship with gastric mucosal inflammation. The results showed that H. pylori and G. hominis could be easily observed in the lower third of the mucous layer and in the mucosa of the gastric pits by means of toluidlne blue staining. Both bacteria immunostained positive. Helicobacter pylori were usually in the shape of curved bacillary while G. hominis often appeared in spiral configuration. In 257 cases of Thai subjects, 169 cases were found to be H. pylori positive, the detection rate was 65.7%, and 16 cases were G. hominis positive, with a 6.2% detection rate. In G. hominis infection, 43.6% of cases had normal gastric mucosa. Superficial, erosive and atrophlc gastritis cases were 13.2, 10.9 and 12.5%, respectively. Mucosal inflammation was usually severe in H. pylori, but neutrophil polymorph infiltration was often mild and focal in G. hominis Infection. Although no G. hominis infection with carcinoma was shown in our cases, the occurrence of mucosal atrophy, metaplasia and dysplasia was higher in both bacterial infections compared with H. pylori‐ and G. hominis‐negative cases. It is suggested that G. hominis may be partly responsible for the mucosal inflammation and some malignant‐associated lesions.

Quantitative study of local distribution of noncholesterol sterols and cholesterol in gallstones

Quantitative analysis of the local distribution of four noncholesterol sterols, 24-methylene cholesterol, campesterol, stigmasterol, and β-sitosterol, and of the local distribution of cholesterol in gallstones was performed by mass spectrometry, with D6-cholesterol as an internal standard. The role played by trace amounts of these four noncholesterol sterols in the formation of gallstones was investigated by comparing the amounts of these sterols in different parts of gall-stones. It was found that the amounts of the noncholesterol sterols in the inside part were significant greater than the amounts in the outside part of various structural types of gallstones. However, the distribution of the cholesterol did not show such variation. The amounts of noncholesterol sterols distributed locally suggested that these sterols play a role in the formation of gallstones.

Recent advance in diagnosis and treatment of cholelithiasis in special reference to ultrasonic tomography

SummaryThe review of 1,041 cases of cholelithiasis operated on during the period of 1935 to 1970 has led to the following conclusion. In diagnosis and treatment of cholelithiasis, careful scrutiny of patient’s history and clinical symptoms and combined use of diagnostic procedures such as cholecystography and ultrasonic tomography are important as well as operative intervention without undue delay. Cholecystography has established itself as the most useful tool in diagnosis of cholelithiasis and has been found to be useful in about three quarter of cases. The presence of intestinal gas in right upper quadrant in duodenum and/or in colon on negative cholecystogram points strongly the presence of acute cholecysttitis and necessitates immediate operative intervention. Ultrasonic tomography has been found to have a great value when cholecystography fails i.e., in case of non-visualisation of biliary tract. Increasing use of these diagnostic procedures in recent years led to the early diagnosis and treatment of cholelithiasis and to a considerable decrease in mortality rate to as low as 1.2 per cent.

Abstract 3834: Difference of host immunity and mucosal defense to H. pylori infection caused Asian paradox on incidence of gastric cancer: Correlation with mucosal mRNA expression of IL-8, pepsinogenI/II ratio, and H. pylori cagA gene mutation in Thai and Japanese gastric cancer

Proceedings: AACR 101st Annual Meeting 2010‐‐ Apr 17‐21, 2010; Washington, DC Background: Recent in vitro studies showed the association between mucosal mRNA expression of IL-8 with cagA, East Asian type in gastric cancer that frequently found in Japanese. In Thais, not only gastric cancer, but also gastroduodenal ulcer incidences are opposite to those in Japanese. We hypothesized that an associations difference between Thais and Japanese stomach mucosal tissue cytokine response to H. pylori infection may lead us to find key role of host defense in gastric carcinogenesis. Method: There were 143 non-cancer volunteers in Thais, and 110 Thai and Japanese gastric cancer patients who underwent endoscopy and gastric surgery. IL-8 cytokine mRNA expression was conducted by real time RT-PCR. Histopathology, serum IgG antibody for H. pylori, pepsinogen I and II were tested. Tissue H. pylori DNA extracted from antral position in stomach was genotyped for cagA mutation. p value <0.05 was considered as a statistically significant. Result: No distribution difference by sex, age, and stage of advance cancer in both nations. H. pylori positivity in Thai gastric cancer was 70.5% (55/78). There is a significant lower PGI/II ratio in Japanese than in Thai gastric cancer, p = 0.026. Thai gastric cancer patients had significantly lower level of PGII/II ratio at means of 3.3±1.7 than in normal one, p =0.042, and of other non-chronic active gastritis (CAG), p =0.002. Means level of IL-8 mRNA expression in Thai cancer and Japanese cancer were 8511.36±5.22 and 174.54±329.51, respectively. Means level of IL-8 mRNA expression in Thai cancer group is also higher than in non-cancer group, p = 0.058. In subgroup analysis, we could not see a significantly differences of IL-8 mRNA expression or PGI/II ratio in positivity and true negativity H. pylori infection. However, in positive cases with less than 2.0 of PGI/II low ratio showed a significant higher level of IL-8 expressions than with PGI/II ratio over 3.0, p < 0.05. In Thai non-cancer, cagA genotype yield of 33 per cent each of Eastern, Western, and Mix type. Low PGI/II ratio did not associate to CAG severity score in Thais. There was only 32/55 (58%) who had positive H. pylori by PCR and pathology with a rising up of H. pylori IgG antibody. In the 6-year followed up of Thai CAG group with H. pylori cagA, Eastern infection, no one develop to be cancer. Conclusion: In this vivo study, we postulated that IL-8 cytokine expression is a predictive marker for poor prognosis incidence in gastric cancer. There is also association to low PGI/II ratio. The opposite result on IL-8 mRNA expression between both nations reflex to individualized host stomach mucosal defense. High level IL-8 mRNA expression in Thais showed mucosal defense in gastric cancer carcinogenesis. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 3834.

Proceedings Of The 21St Autumn Meeting From October 15th-17th, 1979-Maebashi, Japan

When the peripheral blood lymphocytes from patients with various types of hepatitis were stimulated in vitro with liver specific protein, lymphocyte transformation and MIF production were detectable in many cases, especially in chronic active hepatitis. The macrophage activating factor (MAF), a kind of lymphokines, was also detected in the culture medium of activated lymphocytes from patients who showed positive blastogenesis. The activated macrophages by MAF were shown to be cytotoxic to the separated liver cells causing the marked inhibition of albumin synthesis. MAF-containing culture supernatants of these active lymphocytes activated guinea pig macrophages which inhibited the albumin biosynthesis of the isolated liver cells. These observations suggest that the macrophagemediated cytotoxicity may play and role in pathogenesis of chronic active hepatitis.

Studies on gallstones by means of the x-ray diffractometer

Powder X-ray diffractometer char t s of 7 crit ical gal ls tones are shown in F igure 1, where some differences between cholesterol stone and calcium bi l i rubinate stone are confirmed at 20 in 14.9 ~ 15.5 ~ 17 ~ and 23.1 ~ Hydrous cholesterol stone associated by cholesterol shows a spli t t ing peak a t 20 in 14.6 ~ and 14.9, where the fo rmer peak is s t ronger than the la t te r of normal cholesterol stone. A calcium carbonate stone shows an indicant peak at 20 in 29.3 ~ and a fa t ty acid stone gives a dis t inct peak a t 20 in 13.8 ~ 419 206 diffraction charts of gallstone are shown in Table 1, where variations of peak in intensity ~re found. Figure 1. X-Ray Diffractometer Charts of Gallstones

Studies on cholelithiasis

Summary1)The results of the cholecystography in 227 cases with cholelithiasis, using Biloptin and Biligrafin in combination were as follows; in 96 cases of total cases (42.3%) stones were radiologically imaged, in 59 cases (26.0%) no stone was imaged but some abnormal findings in the morphology of the gall bladder or bile ducts were observed and 72 cases (31.7%) were radiologically so called “negative cases.”2)In the radiologically “negative cases”, the differential diagnosis is needed in consideration of other diseases in epigastrium, in which the image of gas formation of the intestinal canals in the right epigastrium plays an important role. In our study, the gas formation was observed in 66 of 72 radiologically negative cases (91.7%) and 34 cases of them were acute gangrenous cholecystitis.3)Cholecystographic studies were also made in dogs which were made artificially allergic cholecystitis in application of the antigen-antibody reaction using the cell substance of coli bacilli; the radiological image of the gall bladder became less and less distinguishable on the time passing until in 72 hours from the beginning it turned to the negative case. la these cases, also the gas formation of the intestinal canals was apparently observed and in these experiments the correlations among cholecystitis, radiologically negative cases and the appearance of the gas formation of the intestinal canals in the right epigastrium were clarified.4)The “Supersonogram”—an ultrasonic wave method in diagnosis—, exploited in our department, made it possible to obtain the recording of the tomogram of the living body and in this method the authors could also confirm the existence of gall stones, the localization, and the depth from the body surface. The stone-echo was obtained in 63 of 68 cases with cholelithiasis (92.6%) and it was a noticeable good result that in 27 of 31 radiologically negative cases (87.1%) the existence of stones was confirmed by “Supersonogram”. In our study it was apparent that the diagnostic result of cholelithiasis could be markedly improved by paying attention to the appearance of the abnormal gas formation of the intestinal canals in the right epigastrium radiologically on the one hand, and applying the “Supersonogram” on the other. The authors could obtain correct diagnosis of cholelithiasis, in 99.1% preoperatively by combining the previously mentioned methods and also in consideration of the clinical symptoms.1) The results of the cholecystography in 227 cases with cholelithiasis, using Biloptin and Biligrafin in combination were as follows; in 96 cases of total cases (42.3%) stones were radiologically imaged, in 59 cases (26.0%) no stone was imaged but some abnormal findings in the morphology of the gall bladder or bile ducts were observed and 72 cases (31.7%) were radiologically so called “negative cases.” 2) In the radiologically “negative cases”, the differential diagnosis is needed in consideration of other diseases in epigastrium, in which the image of gas formation of the intestinal canals in the right epigastrium plays an important role. In our study, the gas formation was observed in 66 of 72 radiologically negative cases (91.7%) and 34 cases of them were acute gangrenous cholecystitis. 3) Cholecystographic studies were also made in dogs which were made artificially allergic cholecystitis in application of the antigen-antibody reaction using the cell substance of coli bacilli; the radiological image of the gall bladder became less and less distinguishable on the time passing until in 72 hours from the beginning it turned to the negative case. la these cases, also the gas formation of the intestinal canals was apparently observed and in these experiments the correlations among cholecystitis, radiologically negative cases and the appearance of the gas formation of the intestinal canals in the right epigastrium were clarified. 4) The “Supersonogram”—an ultrasonic wave method in diagnosis—, exploited in our department, made it possible to obtain the recording of the tomogram of the living body and in this method the authors could also confirm the existence of gall stones, the localization, and the depth from the body surface. The stone-echo was obtained in 63 of 68 cases with cholelithiasis (92.6%) and it was a noticeable good result that in 27 of 31 radiologically negative cases (87.1%) the existence of stones was confirmed by “Supersonogram”. In our study it was apparent that the diagnostic result of cholelithiasis could be markedly improved by paying attention to the appearance of the abnormal gas formation of the intestinal canals in the right epigastrium radiologically on the one hand, and applying the “Supersonogram” on the other. The authors could obtain correct diagnosis of cholelithiasis, in 99.1% preoperatively by combining the previously mentioned methods and also in consideration of the clinical symptoms. The results of the cholecystography in 227 cases with cholelithiasis, using Biloptin and Biligrafin in combination were as follows; in 96 cases of total cases (42.3%) stones were radiologically imaged, in 59 cases (26.0%) no stone was imaged but some abnormal findings in the morphology of the gall bladder or bile ducts were observed and 72 cases (31.7%) were radiologically so called “negative cases.” In the radiologically “negative cases”, the differential diagnosis is needed in consideration of other diseases in epigastrium, in which the image of gas formation of the intestinal canals in the right epigastrium plays an important role. In our study, the gas formation was observed in 66 of 72 radiologically negative cases (91.7%) and 34 cases of them were acute gangrenous cholecystitis. Cholecystographic studies were also made in dogs which were made artificially allergic cholecystitis in application of the antigen-antibody reaction using the cell substance of coli bacilli; the radiological image of the gall bladder became less and less distinguishable on the time passing until in 72 hours from the beginning it turned to the negative case. la these cases, also the gas formation of the intestinal canals was apparently observed and in these experiments the correlations among cholecystitis, radiologically negative cases and the appearance of the gas formation of the intestinal canals in the right epigastrium were clarified. The “Supersonogram”—an ultrasonic wave method in diagnosis—, exploited in our department, made it possible to obtain the recording of the tomogram of the living body and in this method the authors could also confirm the existence of gall stones, the localization, and the depth from the body surface. The stone-echo was obtained in 63 of 68 cases with cholelithiasis (92.6%) and it was a noticeable good result that in 27 of 31 radiologically negative cases (87.1%) the existence of stones was confirmed by “Supersonogram”. In our study it was apparent that the diagnostic result of cholelithiasis could be markedly improved by paying attention to the appearance of the abnormal gas formation of the intestinal canals in the right epigastrium radiologically on the one hand, and applying the “Supersonogram” on the other. The authors could obtain correct diagnosis of cholelithiasis, in 99.1% preoperatively by combining the previously mentioned methods and also in consideration of the clinical symptoms.

Pathophysiological role of bile in the acute intestinal obstruction

In this paper, the resul t s of clinical and exper imenta l observat ions concerning the steroid induced ulcer, are presented. The incidence of the s teroid induced ulcer in the pat ients wi th var ious disease t rea ted in our clinic wi th cort icosteroids, was 2.4%. The incidence of ulcer in all was 2.8%. No difference was found in the incidence of ulcer between the s teroid t reated pa t ien ts and the control ones. Exper imental observat ions of s teroid ulcer were per formed on the guinea pigs. The an imals were feeded normally. 1 or 2 mg of be tamethasone was injected subcutaneously everyday in the periods ranging f rom 14 to 120 days. The s tomachs were fixed in Bouins fluid, embedded in paraffin and s tained with hematoxylin-eosin. Erosions of the mucous membrane occurred only in 3 of 20 guinea pigs.

Immunoserogical studies on the production of cholecystitis

Follow up resu l t s of 45 pat ients in th i s series have been presented. All 5 pa t ien ts wi th heredi tary spherocytosis and acquired hemolytic anemia were cured and gained in good heal th . Two cases wi th idiopathic thrombocytpenic purpura obtained only remission. Results of splenectomy in 4 cases of congestive splenomegaly due to cirrhosis oi the l iver and 2 cases of unknown causes were un i formly poor, The chief cause of death was hematemesis . In early and moderately advanced 32 cases of Banff s syndrom obtained in very good results, and gained in goog heal th of long durat ion. The cause of deaths were not hematemesis bu t mainly c i r rhosis of the liver.