The effects of administration of the Lactobacillus gasseri strain CP2305 on quality of life, clinical symptoms and changes in gene expression in patients with irritable bowel syndrome.

AIMS: To clarify the effects of Lactobacillus gasseri CP2305 (CP2305) on quality of life and clinical symptoms and its functional mechanisms in patients with irritable bowel syndrome (IBS). METHODS AND RESULTS: After the patients were administered CP2305 daily for 4 weeks, the IBS-severity index score was significantly improved compared with that of the placebo group, and this improvement was accompanied by a reduction in health-related worry and changes in intestinal microbiota. The gene expression profiling of the peripheral blood leucocytes showed that CP2305 treatment significantly up-regulated genes related to eukaryotic initiation factor 2 (EIF2) signalling. Eighty-two genes were down-regulated in IBS patients compared with healthy controls. The expression of 23 of these genes exhibited a CP2305-dependent increase associated with an improvement in IBS severity. The majority of the restored genes were related to EIF2 signalling. CONCLUSIONS: CP2305 administration is a potential candidate therapeutic option for patients with IBS. SIGNIFICANCE AND IMPACT OF THE STUDY: Although probiotics have been proposed to benefit IBS patients, objective clinical evidence and elucidation of the functional mechanism remain insufficient. Our study demonstrated that CP2305 administration beneficially influences IBS patients in both subjective and objective evaluations, and gene expression profiling provided insights into the functional mechanism.

Vitamin K3 attenuates lipopolysaccharide-induced acute lung injury through inhibition of nuclear factor-kappaB activation.

Vitamin K is a family of fat-soluble compounds including phylloquinone (vitamin K1), menaquinone (vitamin K2) and menadione (vitamin K3). Recently, it was reported that vitamin K, especially vitamins K1 and K2, exerts a variety of biological effects, and these compounds are expected to be candidates for therapeutic agents against various diseases. In this study, we investigated the anti-inflammatory effects of vitamin K3 in in vitro cultured cell experiments and in vivo animal experiments. In human embryonic kidney (HEK)293 cells, vitamin K3 inhibited the tumour necrosis factor (TNF)-alpha-evoked translocation of nuclear factor (NF)-kappaB into the nucleus, although vitamins K1 and K2 did not. Vitamin K3 also suppressed the lipopolysaccharide (LPS)-induced nuclear translocation of NF-kappaB and production of TNF-alpha in mouse macrophage RAW264.7 cells. Moreover, the addition of vitamin K3 before and after LPS administration attenuated the severity of lung injury in an animal model of acute lung injury/acute respiratory distress syndrome (ARDS), which occurs in the setting of acute severe illness complicated by systemic inflammation. In the ARDS model, vitamin K3 also suppressed the LPS-induced increase in the serum TNF-alpha level and inhibited the LPS-evoked nuclear translocation of NF-kappaB in lung tissue. Despite marked efforts, little therapeutic progress has been made, and the mortality rate of ARDS remains high. Vitamin K3 may be an effective therapeutic strategy against acute lung injury including ARDS.

Retrospective study of technical aspects and complications of endoscopic submucosal dissection for laterally spreading tumors of the colorectum.

BACKGROUND AND STUDY AIMS: Laterally spreading tumors - non granular type (LST-NG) are more often considered candidates for endoscopic submucosal dissection (ESD) than laterally spreading tumors - granular type (LST-G), because of their higher potential for submucosal invasion. However, ESD for LST-NG can be technically difficult. The aim of our study was to compare our ESD results for LST-NG and for LST-G. PATIENTS AND METHODS: Ninety-nine LST-NG and 169 LST-G measuring 20 mm in size or more were removed by ESD. We retrospectively evaluated the clinicopathological features of the tumors and treatment results (en bloc resection rate, procedure time and speed, rate of use of ancillary devices, and complication and recurrence rates). RESULTS: Histopathology revealed that there were more submucosally invasive lesions in the LST-NG than in the LST-G group (28 % vs. 9 %; P < 0.0001). The en bloc resection rate, en bloc R0 resection rate, and en bloc curative resection rate of LST-NG were similar to those of LST-G (LST-NG: 99 %, 98 %, and 88 %; LST-G: 99 %, 98 %, and 91 %). In LST-NG, the median procedure time tended to be longer (LST-NG: 69 min; LST-G: 60 min) and the median procedure speed was slower (LST-NG: 0.15 cm (2)/min; LST-G: 0.25 cm (2)/min; P < 0.0001). Use of ancillary devices was higher for LST-NG (38 % vs. 15 % for LST-G; P < 0.0001), as was the perforation rate (5.1 % vs. 0.6 % for LST-G; P = 0.027). No recurrence was seen in either group. CONCLUSIONS: ESD was an effective treatment method for both LST-NG and LST-G. However, the degree of technical difficulty appears higher for LST-NG than for LST-G lesions, as shown by the lower dissection speed and higher perforation rate. ESD for LST-NG should probably be performed by those with significant experience of colorectal ESD.

Review article: Asia-Pacific consensus recommendations on endoscopic tissue acquisition for biliary strictures.

BACKGROUND: Pre-operative tissue diagnosis for suspected malignant biliary strictures remains challenging. AIM: To develop evidence-based consensus statements on endoscopic tissue acquisition for biliary strictures. METHODS: The initial draft of statements was prepared following a systematic literature review. A committee of 20 experts from Asia-Pacific region then reviewed, discussed, and modified the statements. Two rounds of independent voting were conducted to reach a final version. Consensus was considered to be achieved when 80% or more of voting members voted "agree completely" or "agree with some reservation." RESULTS: Eleven statements achieved consensus. The choice of tissue sampling modalities for biliary strictures depends on the clinical setting, the location of lesion, and availability of expertise. Detailed radiological and endoscopic evaluation is useful to guide the selection of appropriate tissue acquisition technique. Standard intraductal biliary brushing and/or forceps biopsy is the first option when endoscopic biliary drainage is required with an overall (range) sensitivity and specificity of 45% (26%-72%) and 99% (98%-100%), and 48% (15%-100%) and 99% (97%-100%), respectively, in diagnosing malignant biliary strictures. Probe-based confocal laser endomicroscopy and fluorescence in situ hybridisation using 4 fluorescent-labelled probes targeting chromosomes 3, 7, 17 and 9p21 locus may be added to improve the diagnostic yield. Cholangioscopy-guided biopsy and EUS-guided tissue acquisition can be considered after prior negative conventional tissue sampling with an overall (range) sensitivity and specificity of 60% (38%-88%) and 98% (83%-100%), and 80% (46%-100%) and 97% (92%-100%), respectively, in diagnosing malignant biliary strictures. CONCLUSION: These consensus statements provide evidence-based recommendations for endoscopic tissue acquisition of biliary strictures.

Effect of different proton pump inhibitors, differences in CYP2C19 genotype and antibiotic resistance on the eradication rate of Helicobacter pylori infection by a 1-week regimen of proton pump inhibitor, amoxicillin and clarithromycin.

AIM: To investigate the effect of different proton pump inhibitors, S-mephenytoin 4'-hydroxylase (CYP2C19) genotype and antibiotic susceptibility on the eradication rate of Helicobacter pylori. METHODS: One hundred and eighty-seven H. pylori-infected peptic ulcer patients were randomly treated with either rabeprazole (10 mg b.d.) or lansoprazole (30 mg b.d.) plus amoxicillin (750 mg b.d.) and clarithromycin (400 mg b.d.) for 1 week. The antibiotic susceptibility and CYP2C19 genotype (extensive or poor metabolizer) were investigated. RESULTS: The eradication rates in the rabeprazole-amoxicillin-clarithromycin (RAC) and lansoprazole-amoxicillin-clarithromycin (LAC) groups were 75% and 69%, respectively, on an intention-to-treat basis, and 80% and 75%, respectively, on a per protocol basis. The eradication rate for clarithromycin-resistant strains was significantly lower than that for clarithromycin-sensitive strains (24% vs. 86%, P < 0.05). For clarithromycin-sensitive strains in the LAC group, there was a tendency for a lower eradication rate in extensive than poor metabolizers. The eradication rate in extensive metabolizers in the RAC group tended to be higher than that in extensive metabolizers in the LAC group (89% vs. 78%, P = 0.079726). CONCLUSIONS: The success of the 1-week proton pump inhibitor-amoxicillin-clarithromycin regimen depends on the susceptibility of H. pylori to clarithromycin. Moreover, differences in CYP2C19 genotype influence the eradication rates of lansoprazole-based therapy, and the rabeprazole-based regimen has an advantage especially in extensive metabolizers.

Serodiagnosis of alveolar hydatid disease by western blotting.

The purpose of this study was to identify species-specific protein components of Echinococcus multilocularis by Western blotting and establish a simple and highly sensitive method for differential serodiagnosis of alveolar hydatid disease (AHD) from cystic hydatid disease. Using Western blotting analyses we demonstrated 2 previously undescribed, species-specific, antigenic components of protoscoleces of E. multilocularis easily detectable with sera from active AHD patients exclusively. The 18 kDa component was recognized by sera from all active AHD patients, whereas the 16 kDa antigen was characteristically detected by patients with advanced lesions. The potential utility of these components for differential serodiagnosis of AHD is discussed.

Is a proton pump inhibitor necessary for the treatment of lower-grade reflux esophagitis?

The efficacy of histamine H2 receptor antagonist (H2RA) and proton pump inhibitor (PPI) therapies in healing reflux esophagitis was compared in a prospective randomized case-control comparative study. A total of 71 patients with grade A to D esophagitis (Los Angeles classification) were given either famotidine 20 mg twice a day (Fam; n = 35) or omeprazole 20 mg once daily (Ome; n = 36) for 8 weeks. Endoscopy was performed to assess healing in 57 patients. Healed patients were followed-up without H2RA or PPI therapy for 3 months. At the end of follow-up, endoscopy was able to be performed in 33 patients. Healing rates for patients in the Fam and Ome groups were 58.6% (17/OFF and 97.4% (27/28), respectively (P < 0.001), and when limited to grade A to B, healing rates were 60.9% (14/23) and 100% (25/25), respectively (P < 0.001). Concerning Helicobacter pylori infection, healing rates for the Fam and Ome groups in H. pylori (+) patients were 90.0% (9/10) and 90.9% (10/11), respectively (P = 1.00). Remission rates in the Fam and Ome groups were 45.0% (91/2)) and 33.3% (6/18), respectively (P > 0.4). In regard to alcohol drinking, remission rates of daily and social drinkers were 7.7% and 42.4%, respectively (P < 0.03). Thus, PPI should be the drug of choice even for healing lower-grade reflux esophagitis, especially in H. pylori (-) patients. Treatment with H2RA may be an alternative choice in H. pylori (+) patients. After healing, most patients cannot sustain remission without maintenance therapy.

A patient with rectal cancer associated with ulcerative colitis in whom endoscopic ultrasonography was useful for diagnosis.

Endoscopic ultrasonography (EUS) was helpful for the diagnosis of rectal cancer associated with ulcerative colitis. The patient was a 38-year-old Japanese man with a 19-year history of relapsing-remitting type ulcerative colitis involving the entire colon. Routine colonoscopy revealed multiple polypoid prominences in the upper portion of the rectum. EUS revealed a hypoechoic mass in the submucosa beneath and around the polypoid lesion on the most oral side. Signet ring cells were found in a biopsy specimen from this lesion. Subtotal colectomy was performed. A depressed lesion was observed around the prominence on the most oral side; histologically, this lesion was poorly differentiated mucinous and signet ring cell carcinoma extending into the subserosa. The polypoid lesion on the most anal side was well differentiated adenocarcinoma, which was limited to the mucosa. Our findings suggest that EUS is helpful for detecting invasive cancer associated with ulcerative colitis.

Possible formation of new brood capsule by the previously formed brood capsule in Echinococcus multilocularis metacestodes.

In Echinococcus multilocularis metacestodes obtained from the peritoneal cavity of an experimentally infected jird, cellular accumulations were found not only on the inner surfaces of germinal layers but also on the outer surfaces of brood capsules. These cellular accumulations are believed to represent at incipient form of the brood capsule. It has been thought that brood capsules are produced by the germinal layer, but we speculate that the brood capsule itself, as well as the germinal layer, may have the potential to produce new brood capsules.

[In vitro and in vivo effects of paromomycin sulfate on tapeworm, especially Diphyllobothrium ditremum, and clinical effects on diphyllobothriasis in man].

Plerocercoids of D. ditremum were obtained from the Japanese surfsmelt (Hypomesus pretiosus) and administered to Golden hamsters. The animals were treated with paromomycin sulfate on the 16th day of infection. Anthelmintic effect was imperfection after administration of 530 mg/kg in 4 divided doses. Kamala and bithionol, however, successfully expelled the worm at the doses of 80 and 160 mg/kg and 12.5--70 mg/kg, respectively. The balloon-like structures and the other morphological changes of the tegument such as exfoliation of microtriches were remarkably observed on the worms immersed in 2% solution of paromomycin sulfate. Four human cases of diphyllobothriasis were treated with paromomycin sulfate at the doses of 46--56 mg/kg. No side effects were observed. Stool examination made 1 month after treatment were negative to the eggs. From these results, an anthelmintic effect of paromomycin sulfate was shown in the concentration of 2% on D. ditremum in vitro and in the dosage of 46--56 mg/kg in diphyllobothriasis lata patients.

[Epidemiology of alveolar hydatid disease (AHD) and estimation of infected period of AHD in Rebun Island, Hokkaido].

An epidemiological study was performed of endemic alveolar hydatid disease (AHD, multilocular echinococcosis), Rebun Island, Hokkaido and the period of AHD infection of patients was estimated. Death certificates of the residents of the island were analyzed, and 74 deaths (43 males and 31 females) by AHD were found out of the 3,126 deaths that occurred during the period from 1948 to 1990. The red fox population of the island was estimated on the basis of past researchs. The deaths due to AHD distributed around a major peak (n = 67) between 1948-1975 and there were 7 sporadic cases between 1976-90. The red fox population on the island had been estimated to be largest in 1935. The mean infection period from initial AHD infection to death was estimated to be 26 +/- 7 years (x +/- SD) on the basis of the period between the year in which the peak red fox population was observed (1935) and the major peak of patient death (1962). The mean symptomatic period was 5 +/- 5 years, and the mean latent period from infection to the onset of AHD was 21 +/- 7 years. Sex ratio (M/F = 28/13 = 2.15) was higher (P < 0.05) at the age groups below 10 and 26-45 years than the other age groups (15/18 = 0.83), and playing outdoors during childhood and working outdoors in the prime of life were assumed to be the causes of infection.

[Calcium-dependent signaling of acid secretion in isolated parietal cells from guinea pigs and its modification by ethanol].

Treatment of isolated parietal cells from guinea pig gastric mucosa with ethanol caused a rapid increase in [Ca2+]i and concomitant decrease in the capacity for carbachol-stimulated acid secretion in a dose dependent manner. Carbachol rapidly increased the [Ca2+]i from trimethoxybenzoic acid 8-(diethylamino)-octyl ester sensitive intracellular pool. In contrast, the increase with ethanol was through La3+ sensitive Ca2+ channel from external source, which suppressed the Ca2+ response subsequently stimulated with carbachol. Pretreatment of the cells with EGTA or La3+ completely prevented the elevation of [Ca2+]i with ethanol and preserved the Ca2+ response to carbachol. These findings indicate that ethanol-induced elevation of [Ca2+]i may desensitize the stimulation of carbachol. Furthermore, treatment of the parietal cells with ethanol increased the activity of protein kinase C in both cytosolic and membrane fractions of the cells. Activation of protein kinase C with phorbol diester suppressed the capacity for acid secretion. These results suggest that ethanol may inhibit the carbachol-stimulated acid secretion through the desensitization of Ca2+ response and the activation of protein kinase C.

[Risk factors for gastrointestinal bleeding].

Gastrointestinal bleeding sometimes causes life-threatening state. It is important to understand the underlining risk factors for prevention and treatment of this condition. In 1997, 81 patients with massive gastrointestinal bleeding were admitted to the life-saving center in Kyoto First Red Cross Hospital. In these patients, 14 subjects (17%) had been receiving hemodialysis. Eight patients (10%) were taking anti-coagulant or antiplatelet drugs. Eight patients (10%) had hypertension and were given calcium antagonists. Seven subjects (9%) had liver cirrhosis and/or hepatocellular carcinoma. Because these patients often fall into life-threating state, we must pay special attention to the prevention and cure for gastrointestinal bleeding. For example, it may be necessary to change to heparin free hemodialysis for patients having active bleeding. In anticoagulated patients, it may be required that sufficient hemostatic therapy without risking thromboembolic sequelae. In addition to careful managements, we have better to consider the eradication therapy for all of these high risk groups with Helicobacter pylori infection.

[Studies on the diagnosis of schistosomiasis japonica. (1) Circumoval precipitin (COP) test as a method of screening of the subjects for fecal examination (author's transl)].

Examinations were performed on 351 individuals of Shikishima village, Yamanashi Prefecture. Three fecal samples were collected at one-week intervals and examined by MIFC method without iodine. Reactions with a diameter of 9 mm or more in wheal and/or 20 mm or more in erythema were regarded as the positive for the intradermal reaction using Melcher's antigen (1 : 1,000). For the COP test, each one drop of serum and a saline suspension of eggs were mixed on a microscope slide and sealed a coverslip with vaseline. The number of eggs showing precipitates were counted after 48 hrs at 37 degrees C. The results showed that all the precipitates were small in size in the COP titer less than 40%. Medium precipitates appear first in the cases of a 40.% class and the percentage increased thereafter as the COP titer increased. Large precipitate was found in 10% of the eggs affected in a 50% class and found in 40% in the titer of a 80% class. Of 351 tested, a significant higher rate of egg-passers (42.7%, 41/96) was obtained in the COP reactors, as compared with other two groups; 19.4% (41/211) in the dermal reactors, 11.7% (41/351) in the total subjects. Moreover, the egg-passers were recovered in 60--70% of the cases in the COP titer of more than 70%. It was considered that the COP test was a useful diagnostic method for schistosomiasis japonica and the sensitivity and efficiency of this test were superior to those of intradermal test with regarded to the screening of the subjects for fecal examination.

[Enzyme-linked immunosorbent assay (micro-ELISA) for serological diagnosis of multilocular echinococcosis].

Multilocular echinococcosis caused by the larval stage of Echinococcus multilocularis is an important zoonosis with high mortality. In Hokkaido, Japan, although the prevalence among inhabitants has been very low, the areas infested with the parasite are rapidly spreading in recent years. Therefore, we improved on the procedure of the enzyme-linked immunosorbent assay (micro-ELISA) for serological diagnosis of multilocular echinococcosis to carry out the epidemiological survey. A micro-ELISA plate (M129A, DYNATECH), antigen of E. multilocularis cyst extract, peroxidase-conjugated anti-human IgG (MILES-YEDA), O-phenylenediamine and a plate reader (Multiskan MC, TITERTEK) were used for the assay. Reactions were performed with 0.1 ml of reagents in wells of plate at each step and 0.1 ml of 0.2N H2SO4 was finally added for stop of the reaction. The optimum condition was determined by block titration, as 10 micrograms/ml of protein for antigen concentration, 1:40 for serum dilution and 1:4,000 for conjugate dilution. In this assay system, 30 serum samples could be examined in one plate. After correcting the absorbance of negative control to 0.300, positive range was defined as more than 0.600 which was twice the value of negative control. Doubtful positive range was defined as between 0.450 and 0.600, and negative range as less than 0.450. According to this criterion, 30 serum samples of proven patients with multilocular echinococcosis were examined. Positive and doubtful positive reactions were found from 20 sera (66.7%) and 7 sera (23.3%), respectively. Three patient sera (10.0%) showed negative reactions.(ABSTRACT TRUNCATED AT 250 WORDS)

[Susceptibility of the Chinese hamster (Cricetulus griseus) to parasitic infection. (2) Experimental infection with Schistosoma japonicum].

Chinese hamsters (Cricetulus griseus Milne-Edwards: CHA colony) were experimentally infected with Schistosoma japonicum cercariae by the percutaneous route and intraperitoneal inoculation to confirm their susceptibility to the infection with this parasite in comparison with that of mice (DD). Chinese hamsters proved to be highly susceptible to percutaneous and intraperitoneal infection with S. japonicum. The eggs of S. japonicum were recognized in feces of all Chinese hamsters and mice from about 6 weeks after infection. The mortality within 50 weeks post-infection was lower in Chinese hamsters than in mice. Eight out of 18 Chinese hamsters survived more than 50 weeks post-infection, seven of these showed eggs in their feces beyond this period, whereas no fecal eggs was detected in the same period in the five mice which survived more than 50 weeks post-infection. Chinese hamsters seem to be equally susceptible as mice to S. japonicum infection and more tolerant than mice to the worm load.

[Susceptibility of the Chinese hamster (Cricetulus griseus) to parasitic infections. (1). Experimental infection with Nematospiroides (Heligmosomoides) dubius to the cortisone-treated Chinese hamster].

Chinese hamsters (Cricetulus griseus) were experimentally infected with Nematospiroides (Heligmosomoides) dubius. Though they were resistant to the infection of this parasite, they were found to become susceptible to it by the treatment with cortisone. The number of the worms in the intestine of the Chinese hamster and the number of the eggs oviposited by female worms were markedly affected by the doses of the cortisone. Egg oviposition of N. (H.) dubius ceased within 27 days post infection in the cortisone treated Chinese hamster, probably because of the expulsion of the worms. It was suggested that cortisone treatment controlled the infection of the Chinese hamster with N. (H.) dubius and the reproductive activity of this parasite.

[Susceptibility of the Chinese hamster (Cricetulus griseus) to parasitic infection (3). Experimental infection with Hymenolepis nana or Trichuris muris to the cortisone treated Chinese hamster].

Susceptibility of Chinese hamster (Cricetulus griseus) of Asahikawa Colony (CHA) to Hymenolepis nana or Trichuris muris infection was compared in the feces-egg examination with that of mice as the control animals. Though CHA were resistant to the infection of H. nana, they were found to become susceptible to H. nana by the treatment with cortisone. A half number of CHA was infected with H. nana and the eggs were detected from each animal only in 4 or 6 days in the periods of examination more than 40 days. Mice with or without cortisone treatment were equally susceptible to H. nana infection. In another experiment, CHA with or without cortisone treatment were completely resistant to Trichuris muris infection. Mice, as the control animals, were found to be infected with T. muris in both of cortisone-treated and non-treated groups. Results from the fecal examination, it was confirmed that T. muris were expelled naturally from the animals on the weeks of 11 to 33 after infection.

[Detection of echinococcal infection in rodents and shrews caught in Asahikawa and Kushiro districts].

During the May-October period of 1983, rodents and shrews were caught in Asahikawa and Kushiro districts and examined for the presence of echinococcal infection. 603 individuals representing 8 species and 110 individuals of 9 species were collected in Asahikawa and Kushiro districts, respectively. Among these animals, a vole, Clethrionomys rutilus mikado caught in Akkeshi in Kusiro district was infected with larval Echinococcus multilocularis, but no evidence of echinococcal infection was found in the animals caught in Asahikawa.