High incidence of autoimmune gastritis in patients misdiagnosed with two or more failures of H. pylori eradication.

BACKGROUND: Although autoimmune gastritis (AIG) is generally considered relatively rare, we frequently encounter AIG among patients at to our hospital who have experienced at least two episodes of Helicobacter pylori eradication failure. AIMS: We investigated the incidence of AIG in consecutive patients who consulted our department for H. pylori eradication with reference to eradication history. METHODS: A total of 404 consecutive patients who visited the H. pylori-specific out-patient unit of our hospital from June 2015 to June 2017 were enrolled. Of these, 137 were treatment-naive, 47 had failed treatment once (single failure), and 220 had failed treatment twice or more (multiple failures) by 13 C-UBT. Gastroscopy was performed in all patients. Culture tests of gastric mucosal samples were performed for H. pylori and other bacteria positive for urease activity. Anti-parietal cell antibody (APCA) was measured. Patients with severe atrophy in the gastric corpus and positivity for APCA were diagnosed as having AIG. RESULTS: A total of 43 patients were diagnosed as having AIG, of whom two were treatment-naive (1.5%, 2/137), 1 failed eradication once (2.1% 1/47), and 40 failed treatment at least twice (18.2%, 40/220). The incidence of AIG was significantly higher in the multiple failure group than in the single failure or treatment-naive groups. Urease-positive bacteria, such as Klebsiella pneumoniae and alpha-streptococcus, were identified in 33 of the 35 AIG patients who underwent culture testing. CONCLUSION: AIG patients were often misdiagnosed as refractory to eradication therapy, probably because achlorhydria in AIG might allow urease-positive bacteria other than H. pylori to colonise the stomach, causing positive 13 C-UBT results.

Potent acid inhibition by vonoprazan in comparison with esomeprazole, with reference to CYP2C19 genotype.

BACKGROUND: Acid inhibitory effects of proton pump inhibitors (PPIs) are influenced by CYP2C19 genotype. In contrast, the potent acid inhibition of vonoprazan is not influenced by CYP2C19 genotype. AIM: To compare the acid inhibitory effects of vonoprazan and esomeprazole in relation to CYP2C19 genotype. METHODS: Twenty-eight healthy Japanese volunteers [7 CYP2C19 poor metabolisers (PMs), 11 intermediate metabolisers (IMs) and 10 rapid metabolisers (RMs)] received four different regimens in a randomised crossover manner: (i) vonoprazan 20 mg twice daily (b.d.), (ii) vonoprazan 20 mg daily, (iii) esomeprazole 20 mg b.d. and (iv) esomeprazole 20 mg daily. The timing of each dosing was 1 h before a meal. Twenty-four-hour intragastric pH monitoring was performed on day 7 on each regimen. RESULTS: In the overall genotype group, pH ≥4 holding time ratios (pH 4 HTRs) with vonoprazan b.d., vonoprazan daily, esomeprazole b.d. and esomeprazole daily were 100%, 95%, 91%, and 68% respectively. pH 5 HTRs were 99%, 91%, 84% and 54% respectively. Vonoprazan b.d. potently suppressed acid for 24 h, and was significantly superior to other regimens irrespective of CYP2C19 genotype. Vonoprazan daily was equivalent to esomeprazole b.d. in IMs and PMs, but superior in RMs. CYP2C19 genotype-dependent differences were observed in esomeprazole daily but not in vonoprazan b.d. or daily. CONCLUSION: Vonoprazan 20 mg b.d. inhibits acid irrespective of CYP2C19 genotype, more potently than esomeprazole 20 mg b.d., pH 4 and 5 holding time ratios reached 100% and 99%, respectively.

Role of aquaporin-5 in gallbladder carcinoma.

BACKGROUND/PURPOSE: Aquaporins (AQPs) are important in controlling bile formation. However, the exact role in human gallbladder carcinogenesis has not yet been defined. METHODS: AQP-5-expressing gallbladder carcinoma (GBC) cell lines (NOZ) were transfected with anti-AQP-5 small interfering RNA (siRNA). Growth, migration, invasion assay, and drug susceptibility tests were performed. Next, microRNA (miRNA) expression was analyzed by miRNA oligo chip (3D-Gene®). AQP-5 and AQP-5-related miRNA target gene expressions were also analyzed using tissue microarray (TMA) in 44 GBC samples. RESULTS: Treatment with AQP-5 siRNA decreased cell proliferation, migration, and invasion. On the other hand, those cells increased IC50 of gemcitabine. By performing miRNA assays, miR-29b, -200a, and -21 were shown to be highly overexpressed in cells treated with AQP-5 siRNA NOZ. When focusing on miR-21, phosphatase and tensin homolog (PTEN) was found to be a target of miR-21. In the TMA, AQP-5/PTEN coexpression was significantly associated with the depth of invasion and MIB-1 index (p = 0.003, 0.010). Survival of patients with a high AQP-5/PTEN coexpression was longer than that of patients with a low coexpression (p = 0.003). CONCLUSIONS: Our result suggested that miR-21 and PTEN may contribute to the role of AQP-5 in GBC. AQP-5 and PTEN cascades are favorable biomarkers of GBC.

Twice-daily dosing of esomeprazole effectively inhibits acid secretion in CYP2C19 rapid metabolisers compared with twice-daily omeprazole, rabeprazole or lansoprazole.

BACKGROUND: Twice-daily dosing of proton pump inhibitors (PPIs) is used to treat Helicobacter pylori or acid-related diseases, such as gastro-oesophageal reflux disease (GERD) refractory to standard dose of a PPI. Genetic polymorphisms of CYP2C19 are involved to different extents in the metabolism of four kinds of PPIs (omeprazole, lansoprazole, rabeprazole and esomeprazole) available in Japan. AIM: To compare acid-inhibitory effects of the four PPIs dosed twice daily in relation to CYP2C19 genotype. METHODS: We performed 24-h pH monitoring studies on Day 7 of PPI treatment for 40 Japanese H. pylori-negative volunteers [15 CYP2C19 rapid metabolisers (RMs), 15 intermediate metabolisers (IMs) and 10 poor metabolisers (PMs)] using a randomised four-way crossover design: omeprazole 20 mg, esomeprazole 20 mg, lansoprazole 30 mg and rabeprazole 10 mg twice daily. RESULTS: Although median pH values with esomeprazole, omeprazole, lansoprazole and rabeprazole were 5.7 (3.5-7.2), 5.5 (2.4-7.2), 5.5 (3.7-7.3) and 5.2 (2.5-7.3), respectively (no statistically significant differences), CYP2C19 genotype-dependent differences were smaller for esomeprazole and rabeprazole compared with values for omeprazole and lansoprazole. In CYP2C19 RMs, the median pH with esomeprazole [5.4 (3.5-6.8)] was significantly higher than those with omeprazole [5.0 (2.4-5.9), P = 0.018], lansoprazole [4.7 (3.7-5.5), P = 0.017] or rabeprazole [4.8 (2.5-6.4), P = 0.002]. In IMs and PMs, the median pH was >5.0 independent of the PPI. CONCLUSIONS: In intermediate and rapid metabolisers of CYP2C19, PPIs dosed twice daily could attain sufficient acid suppression, while in CYP2C19 RMs, esomeprazole 20 mg twice daily caused the strongest inhibition of the four PPIs. Therefore, esomeprazole may be effective in Japanese population when dosed twice daily.

Importance of the six angiosomes concept through arterial-arterial connections in CLI.

AIM: We reviewed the importance of six angiosomes concept in the foot area through arterial-arterial connections. METHODS: We retrospectively analyzed 145 limbs from 111 critical limb ischemia (CLI) patients to investigate whether the wound location corresponded with the occlusion of the feeding artery. We also analyzed 57 limbs that underwent endovascular therapy (EVT). Regardless of which target vessel underwent EVT in the calf area, it was considered "direct" if the feeding artery flow in the foot area was achieved and "indirect" if not achieved. The wound healing rate was compared between the direct group and the indirect group relative to the six angiosomes of the foot. Further, we analyzed the absolute difference of the skin perfusion pressure (SPP) values measured before and after EVT. RESULTS: A concordance rate of 82.1% (119/145 limbs) was observed between the wound location and the site of vessel occlusion. The wound healing rate of the six angiosomes-direct group was significantly higher than that of the six angiosomes-indirect group (96.6% vs. 72.7%; P=0.03). The SPP values were significantly higher in the six angiosomes-direct group than those in the six angiosomes-indirect group (20.3±18.1 mmHg vs. 5±14.5 mmHg; P=0.039). CONCLUSION: In the case of the patency of the arterial-arterial connections, even if one undergoes EVT for any of the three vessels in the lower leg, wound healing is likely to be achieved, if we can achieve six angiosome-direct flow.

Neutralization of the γ-secretase activity by monoclonal antibody against extracellular domain of nicastrin.

Several lines of evidence suggest that aberrant Notch signaling contributes to the development of several types of cancer. Activation of Notch receptor is executed through intramembrane proteolysis by γ-secretase, which is a multimeric membrane-embedded protease comprised of presenilin, nicastrin (NCT), anterior pharynx defective 1 and PEN-2. In this study, we report the neutralization of the γ-secretase activity by a novel monoclonal antibody A5226A against the extracellular domain of NCT, generated by using a recombinant budded baculovirus as an immunogen. This antibody recognized fully glycosylated mature NCT in the active γ-secretase complex on the cell surface, and inhibited the γ-secretase activity by competing with the substrate binding in vitro. Moreover, A5226A abolished the γ-secretase activity-dependent growth of cancer cells in a xenograft model. Our data provide compelling evidence that NCT is a molecular target for the mechanism-based inhibition of γ-secretase, and that targeting NCT might be a novel therapeutic strategy against cancer caused by aberrant γ-secretase activity and Notch signaling.

Course of the bony canal associated with high-positioned supraorbital foramina: an anatomic study to facilitate safe mobilization of the supraorbital nerve.

OBJECTIVE: In the dissection of the superior orbital rim, the supraorbital foramen must be released to preserve the supraorbital nerve. The aim of this study was to clarify the spatial dimensions of a high-positioned foramina to allow for the safe performance of this maneuver. METHODS: We examined 90 orbital sides. In the detected foramina we measured the distance between the superior orbital rim and the inferior margin of the foramen (height), and between the rim and the anterior margin of the opening of the foramen in the orbital roof (depth). To evaluate the inclination of the canal, we calculated the height:depth ratios. Foramina with a height exceeding 2.0 mm were defined as high-positioned and measurements between low- and high-positioned foramina were compared. RESULTS: We were able to find 37 foramina in 32 orbital sides; 25 (67.6%) were low-positioned and their height ranged from 0.5-1.9 mm (mean: 1.16 mm); 12 (32.4%) were high-positioned with a height ranged from 2.0-11.9 mm (mean: 4.0 mm). The depth in low- and high-positioned foramina ranged from 0.5-3.2 mm (mean: 1.44 mm) and 0.7-6.5 mm (mean: 2.13 mm), respectively. The height:depth ratio was greater in high-positioned foramina, ranging from 1.03-3.38 (mean: 2.19), than in low-positioned foramina where it ranged from 0.4-2.2 (mean: 0.98). CONCLUSION: The height:depth ratio in high-positioned foramina, an approximate mean value of 2, and the absence of a value less than 1, was considered to indicate a steep canal inclination. To avoid postoperative forehead numbness, anatomic information regarding the course of the bony canal must be considered.

The use of colour Doppler imaging in the diagnosis of retinal detachment.

DESIGN: Prospective study. PURPOSE: To evaluate real-time duplex colour Doppler examination with colour and pulse Doppler ultrasonography (US) (colour Doppler imaging (CDI)) for detection of the retinal detachment from membranous structures in the posterior segment. MATERIALS AND METHODS: In 33 consecutive patients with ophthalmoscopically invisible eyes (34 eyes), CDI was performed to detect the presence or absence of retinal detachment. The diagnostic criterion for retinal detachment was whether or not spectral waveforms were detected in membranous structures with colour and pulse Doppler US. In all cases, the absence or presence of retinal detachment was confirmed by surgery. RESULTS: In 27 of 34 eyes, membranes and/or opacities were observed in the vitreous cavity with CDI. In 12 of these 27 eyes, blood flow in those structures was detected by CDI. In all of these 12 eyes, retinal detachment was confirmed at surgery, and in 14 of the 15 eyes in which blood flow was not detected by CDI, absence of retinal detachment was confirmed at surgery. When a diagnosis confirmed by surgery was used as the definitive finding, CDI had a sensitivity of 92.3%, specificity of 100%, positive predictive value of 100%, negative predictive value of 93.3%, and an accuracy in the detection of retinal detachment from membranes and/or opacities in the vitreous cavity of 96.3%. CONCLUSION: Real-time duplex colour Doppler examination with colour and pulse Doppler US (CDI) is a noninvasive method useful for the detection of retinal detachment from vitreous opacity and/or membrane in the posterior segment.

[Papillary fibroelastoma of the right atrium; report of a case].

Papillary fibroelastoma is a rare benign tumor commonly arising from a heart valve. We describe an unusual papillary fibroelastoma that arose from the right side of the interatrial septum. An intracardiac tumor was discovered by routine echocardiography in an asymptomatic 68-year-old woman. The echocardiographic examination revealed a 20 mm mobile tumor in the right atrium. Tricuspid obstruction was not observed, nor was regurgitation. The tumor was resected through a right atriotomy. It had multiple papillary fronds and arose from the interatrial septum. Pathologic examination confirmed papillary fibroelastoma. The postoperative course was uneventful, and the patient was discharged on postoperative day 13.

[The control survey of Japanese Association of Medical Technologists (JAMT) and its possibility of the standardization for the survey].

External quality control survey of JAMT (Japanese Association of Medical Technologists) carries out 9 sections of clinical chemistry, hematology, bacteriology, serology, cytology, urology, physiology-pathology, blood transfusion and genetics. In two sections of biochemistry and urology, the control survey sample have been certified which was target value and allowance limits of error by the reference method. These standardized items are utilized in order to know the degree of the accuracy of the participation laboratories. We carried out the investigation for putting the focus at 2 sections of biochemistry and serology in order to execute the standardization of the external quality control. The multiple laboratories were carried out certified using both of common pooled serum and the reference serum for a proficiency test. On the standardization items, the result showed that the external quality survey assessment was possible by the accuracy. On items of biochemistry and serology test, there have compatibility which the results of same control survey of other group. It judged possible the standardization of the external quality control assessment by the integration of the research method.

A clinicopathologic study of mucinous adenocarcinoma of the stomach.

BACKGROUND: Mucinous adenocarcinoma of the stomach is a rare histologic type of gastric adenocarcinoma and its features are still controversial. We attempted to clarify the clinicopathologic characteristics of this histologic type. METHODS: We reviewed the records of 112 patients with mucinous adenocarcinoma of the stomach (MUC) and 4160 patients with nonmucinous gastric adenocarcinoma (NMUC) for factors including age; sex; tumor location, size, and depth; lymph node metastasis; lymphatic or venous permeation; peritoneal dissemination; liver metastasis; and survival rate. We also investigated the relationship between cancer depth and lymph node metastasis. Statistical analysis included chi 2 and Student's t-tests. Survival rates were calculated by the Kaplan-Meier method and analyzed by the log rank test. RESULTS: MUC was found mostly in younger patients, in the lower part of the stomach and was of larger size, invading to or beyond the muscularis propria, positive for lymph node involvement, and associated with peritoneal dissemination. The overall 5-year survival rate in patients with MUC was lower than that in NMUC patients, because the MUC patients more frequently had advanced stage disease; however, the survival rates in the two groups did not differ significantly. As for the relationship between cancer depth and lymph node involvement, the frequency of lymph node metastasis increased when invasion reached the submucosa in patients with MUC compared with those with NMUC while the frequency of lymph node metastasis increased in the muscularis propria in patients with NMUC compared with those with NMUC. CONCLUSION: MUC was characteristically found in younger patients, at lower sites, at an advanced stage, and with peritoneal dissemination; MUC had a poor prognosis. Lymph node metastasis should be suspected when MUC invades to the submucosa or deeper.

Catalog of 258 single-nucleotide polymorphisms (SNPs) in genes encoding three organic anion transporters, three organic anion-transporting polypeptides, and three NADH:ubiquinone oxidoreductase flavoproteins.

We present here a series of high-density maps of single-nucleotide polymorphisms (SNPs) detected in genes encoding three organic-anion transporters, three organic anion-transporting polypeptides, and three nicotinamide adenine dinucleotide, reduced:ubiquinone oxidoreductase flavoproteins. A total of 258 SNPs were identified among these nine genes through systematic screening of DNA from 48 Japanese individuals: 17 in 5' flanking regions, three in 5' untranslated regions, 13 in coding regions, 211 in introns, six in 3' untranslated regions, and 8 in 3' flanking regions. By comparing our data with SNPs deposited in the dbSNP database in the National Center for Biotechnology Information, we determined that 236 (91.5%) were novel. In addition, 46 genetic variations of other types were discovered within these loci. These high-resolution maps will serve as a useful resource for analyzing potential associations between variations in these nine genes and differences in human susceptibilities to common diseases or response to drug therapies.

Species-specific differences in expression of G-protein-coupled receptor kinase (GRK) 7 and GRK1 in mammalian cone photoreceptor cells: implications for cone cell phototransduction.

Desensitization plays an important role in the rapid termination of G-protein signaling pathways. This process, which involves phosphorylation by a G-protein-coupled receptor kinase (GRK) followed by arrestin binding, has been studied extensively in the rod photoreceptor cell of the mammalian retina. In contrast, less is known regarding desensitization in cone photoreceptor cells, which occurs more rapidly than in rod cells. Recently, our laboratory has cloned a novel GRK family member, GRK7, from the retina of a cone-dominant mammal, the 13-lined ground squirrel. Here we report the cloning of GRK7 from rod-dominant pig and human retinas, suggesting that this kinase plays a role in human visual signaling. Because GRK1 (rhodopsin kinase), the GRK that mediates rhodopsin desensitization in the rod cell, is reportedly expressed in both rods and cones, a detailed comparison of the localization of the two kinases is a necessary step toward determining their potential roles in cone visual signaling. Immunocytochemical analysis using antibodies selective for these two GRKs unexpectedly demonstrated species-specific differences in GRK7 and GRK1 expression in cones. In pigs and dogs, cones express only GRK7, whereas in mice and rats, we detected only GRK1 in cones. These results suggest that either GRK7 or GRK1 may participate in cone opsin desensitization, depending on the expression pattern of the kinases in different species. In contrast, GRK7 and GRK1 are coexpressed in monkey and human cones, suggesting that coordinate regulation of desensitization by both kinases may occur in primates.

Sympatric convergence of the color pattern in the Chilean Ceroglossus ground beetles inferred from sequence comparisons of the mitochondrial ND5 gene.

The phylogenetic trees have been constructed using the mitochondrial ND5 gene sequences for 66 specimens of the Chilean Ceroglossus ground beetles collected from various localities of Chile. The trees show that the Ceroglossus specimens examined are composed of four phylogenetic lineages that have diverged 25-30 MYA. The results are consistent with the classification proposed by Jiroux (1996) based on morphology, in which Ceroglossus is divided into four species groups. Despite a remarkable color polymorphism revealed by these ground beetles, the color pattern is geographically linked and is not species-specific, suggesting that some ecological or environmental factors are involved in determining it.

Evolutionary discontinuity of the Carabine ground beetles.

The Carabine ground beetles are mostly hind wing-less and cannot fly, so that there is more chance of diversification by geographic isolation compared with winged insects. The relationships between morphological diversification and phylogeny of the ground beetles of the world have been inferred mainly by comparisons of mitochondrial ND5 gene sequences. Based on dating by a mitochondrial DNA "clock," it has been deduced that an explosive radiation of the major carabine groups took place 50-40 MYA. This was followed by occasional radiations on various scales, sometimes accompanied by parallel morphological changes. There are also a good number of examples showing that the fundamental morphology has remained unchanged for a long time among geographically isolated populations within the same species. Thus, carabid evolution would have proceeded discontinuously, with phases of rapid morphological change alternating with silent phases.

Catalog of 46 single-nucleotide polymorphisms (SNPs) in the microsomal glutathione S-transferase 1 (MGST1) gene.

A major goal in our laboratory is to understand the role of common genetic variations among individual patients as regards susceptibility to common diseases and differences in therapeutic efficacy and/or side effects of drugs. As an addition to the high-density SNP (single-nucleotide polymorphism) maps of 12 glutathione S-transferase and related genes reported earlier, we provide here an SNP map of the microsomal glutathione S-transferase 1 (MGST1) gene. Among 48 healthy Japanese volunteers examined. we identified a total of 46 SNPs at this locus, 36 of which had not been reported before: 4 in the promoter region, 34 in introns, 3 in the 3' untranslated region, and 5 in the 3' flanking region. No SNP was found in 5'untranslated or coding regions. The ratio of transition to transversion was approximately 1.2:1. Among the 13 insertion-deletion polymorphisms was a 2-bp deletion in the coding region of MGST1 in DNA from one of the volunteers, which resulted in a frame-shift mutation. Since the gene product encoded by this mutant allele would lack the C-terminal half including the MAPEG (membrane-associated proteins in eicosanoid and glutathione metabolism) domain, MGST1 activity is likely to be reduced in the carrier's cells. The SNP map presented here adds to the archive of tools for studying complex genetic diseases, population migration patterns, and a variety of pharmacogenetic possibilities.

High-density single-nucleotide polymorphism (SNP) map of the 150-kb region corresponding to the human ATP-binding cassette transporter A1 (ABCA1) gene.

Highly dense catalogs of human genetic variations, in combination with high-throughput genotyping technologies, are expected to clarify individual genetic differences in pharmacological responsiveness and predispositions to common diseases. Here we report single-nucleotide polymorphisms (SNPs) present among 48 Japanese individuals at the locus for the human ATP-binding cassette transporter A1 (ABCA1) gene. ABCA1 plays a key role in apolipoprotein-mediated cholesterol transport, and mutations in this gene are responsible for Tangier disease and familial high-density lipoprotein deficiency associated with reduced cholesterol efflux. We identified a total of 162 SNPs, 149 of which were novel, within the 150-kb region encompassing the entire ABCA1 gene. Eight of the SNPs lie within coding elements, two in 5' flanking regions, 147 in introns, and five in 3' untranslated regions, but none were found in 5' untranslated or 3' flanking regions. The ratio of transitions to transversions was approximately 2.37 to 1. Our dense SNP map of this region could serve as a powerful resource for studies of complex genetic diseases that may be associated with ABCA1 and of individual responses to drug therapy.