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International audience; Alterations of mitochondria have been linked to several cancers. Also, the mitochondrial DNA copy number (mtDNA-CN) is altered in various cancers, including gastrointestinal tract (GIT) cancers, and several research groups have investigated its potential as a cancer biomarker. However, the exact causes of mtDNA-CN variations are not yet revealed. This review discussed the conceivable players in this scheme, including reactive oxygen species (ROS), mtDNA genetic variations, DNA methylation, telomere length, autophagy, immune system activation, aging, and infections, and discussed their possible impact in the initiation and progression of cancer. By further exploring such mechanisms, mtDNA-CN variations may be effectively utilized as cancer biomarkers and provide grounds for developing novel cancer therapeutic agents.

Background: Intestinal metaplasia, gastric-to-intestinal transdifferentiation, occurs as a result of the misexpression of certain regulatory factors, leading to genetic reprogramming. Here, we have evaluated the H. pylori-induced expression patterns of these candidate genes.Methods: The expression levels of 1)tissue-specific transcription factors (RUNX3, KLF5, SOX2, SALL4, CDX1 and CDX2), 2)stemness factors (TNFRSF19, LGR5, VIL1) and 3)tissue-specific mucins (MUC5AC, MUC2) were evaluated by quantitative real-time PCR in gastric primary cells (GPCs), in parallel with two gastric cancer (MKN45 and AGS) cell lines, up to 96h following H. pylori infection. Results: Following H. pylori infection of GPCs, RUNX3 declined at 24h post infection (PI) (-6.2±0.3) and remained downregulated for up to 96h. Subsequently, overexpression of self-renewal and pluripotency transcription factors, KLF5 (3.6±0.2), SOX2 (7.6±0.5) and SALL4 (4.3±0.2) occurred. The expression of TNFRSF19 and LGR5, demonstrated opposing trends, with an early rise of the former (4.5±0.3) at 8h, and a simultaneous fall of the latter (-1.8±0.5). This trend was reversed at 96h, with the decline in TNFRSF19 (-5.5±0.2), and escalation of LGR5 (2.6±0.2) and VIL1 (1.8±0.3). Ultimately, CDX1 and CDX2 were upregulated by 1.9 and 4.7-fold, respectively. The above scenario was, variably observed in MKN45 and AGS cells.Conclusion: Our data suggests an interdependent gene regulatory network, induced by H. pylori infection. This interaction begins with the downregulation of RUNX3, upregulation of self-renewal and pluripotency transcription factors, KLF5, SOX2 and SALL4, leading to the downregulation of TNFRSF19, upregulation of LGR5 and aberrant expression of intestine-specific transcription factors, potentially facilitating the process of gastric-to-intestinal transdifferentiation.

Background Amoxicillin-resistant H. pylori strains are increasing worldwide. To explore the potential resistance mechanisms involved, the 3D structure modeling and access tunnel prediction for penicillin-binding proteins (PBP1A) was performed, based on the Streptococcus pneumoniae, PBP 3D structure. Molecular covalent docking was used to determine the interactions between amoxicillin (AMX) and PBP1A. Results The AMX-Ser368 covalent complex interacts with the binding site residues (Gly367, Ala369, ILE370, Lys371, Tyr416, Ser433, Thr541, Thr556, Gly557, Thr558, and Asn560) of PBP1A, non-covalently. Six tunnel-like structures, accessing the PBP1A binding site, were characterized, using the CAVER algorithm. Tunnel-1 was the ultimate access route, leading to the drug catalytic binding residue (Ser368). This tunnel comprises of eighteen amino acid residues, 8 of which are shared with the drug binding site. Subsequently, to screen the presence of PBP1A mutations, in the binding site and tunnel residues, in our clinical strains, in vitro assays were performed. H. pylori strains, isolated under gastroscopy, underwent AMX susceptibility testing by E-test. Of the 100 clinical strains tested, 4 were AMX-resistant. The transpeptidase domain of the pbp1a gene of these resistant, plus 10 randomly selected AMX-susceptible strains, were amplified and sequenced. Of the amino acids lining the tunnel-1 and binding site residues, three (Ser414Arg, Val469Met and Thr556Ser) substitutions, were detected in 2 of the 4 resistant and none of the sequenced susceptible strains, respectively. Conclusions We hypothesize that mutations in amino acid residues lining the binding site and/or tunnel-1, resulting in conformational/spatial changes, may block drug binding to PBP1A and cause AMX resistance.

Background: Quantitation of Helicobacter pylori (Hp) in the gastric tissue is essential for assessment of vaccination/therapeutic regimens. Methods & Results: Here, the inhibitory effect of mouse gastric DNA (MgDNA) on amplification of Hp genomic DNA (HpDNA) was evaluated by spiking HpDNA with serial dilutions of MgDNA, which yielded concentrations of >10 ng/µl and >0.63-10 ng/µl of MgDNA, as inhibition and interference zones, respectively. Mice were then inoculated with varying doses of Hp and assessed at the inhibition-free concentration of 0.63 ng/µl. The average Hp copy numbers per microgram of gastric tissue discriminated mice having received high vs. low dose inoculums (p < 0.001). Secondly, Hp copy numbers were quantitated in immunized mice, which demonstrated significantly lower numbeZrs, in reference to controls (p = 0.006). Conclusion: Our method, bypassing the inhibition and/or interference imposed by MgDNA, was able to quantitate gastric tissue-colonizing Hp, segregating mice inoculated with low vs. high doses of Hp, as well as those immunized from controls.

The rates and routes of Helicobacter pylori transmission, in a high-prevalent country like Iran, with gastric cancer as the leading cause of male cancer mortality, are of great essence. Here, we have studied the H. pylori-associated risk factors and the likelihood of interspousal transmission.In a cohort of 686 young prewed couples, questionnaires were self-administered and serum samples were collected, for assessment of risk factors and H. pylori serostatus, at baseline and follow-up. Of the 475 H. pylori single- or double-seronegative couples, 201 returned for follow-up. The average follow-up duration was 2.2 (SD 0.6) years, with a total of 560.1 person-years. Logistic regression and Cox regression models were used to estimate the odds ratios (ORs) and hazard ratios (HRs).The risk of infection was higher in men than women (OR: 1.3, 95% CI: 1.0-1.8) and among metropolitan than rural residents (OR = 1.4, 95% CI: 1.1-1.9). It was also significantly higher among those with three (OR = 1.6, 95% CI: 1.1-2.2), and four or more siblings (OR = 1.4, 95% CI: 1.0-1.9), in reference to those with one or no siblings. Adult H. pylori acquisition occurred in 10.9% (27/247) of the seronegative participants. The risk of the acquisition was significantly associated with age (P value for trend=0,000). It was also significantly lower among participants who had various degrees of education as compared to illiterate subjects (HR = 0.2, 95% CI: 0.1-0.9). Nevertheless, our analysis did not find any evidence for interspousal transmission (HR = 1.0, 95% CI: 0.4-2.2).Whilst H. pylori acquisition was detected in the young adult Iranian population, our findings did not support interspousal transmission, as a mode of acquisition.

International audience; We have performed a systematic review and meta-analysis for evaluation of mitochondrial DNA copy number (mtDNA-CN) alterations in peripheral blood leukocytes (PBL), and tumor tissues of gastrointestinal tract (GIT) cancers. Analysis of the PBL demonstrated a significant decrease [OR: 0.6 (0.5, 0.8)] and increase [OR: 1.4 (1.1, 1.9)] prior to and following GIT cancer development, respectively. This trend was more evident in CRC, and GC subgroups. Analysis of tissue yielded high levels of heterogeneity. However, the mean difference for the CRC subgroup was statistically significant [1.5 (1.0, 2.2)]. Our analysis suggests mtDNA-CN deserves further investigations as a GIT-cancer screening tool.

Background Food insecurity can increase risks of health and nutritional problems, leading to difficulties in self-care and poor glycemic control in diabetic patients. This study assessed food insecurity and its association with diabetes control and self-care in type 2 diabetes patients. Methods In this cross sectional study, 148 adults with type 2 diabetes participated. Food insecurity and self-care were determined using the 18-item USDA household food security status questionnaire and self-care (SDSCA) questionnaire. Data were analyzed using logistic and linear regression tests (SPSS 20 software). Results Thirty-seven percent of the participants were food-insecure and significantly more likely than food-secure participants to have poor glycemic control (OR = 3.02; CI: 1.45–2.65). No significant association was found between food-insecurity and overall self-care score. Conclusion Food-insecurity was directly associated with poor glycemic control in type 2 diabetes patients. Since economic status was significantly better in the food-secure group, it can be postulated that financial problems will eventually lead to poor glycemic control. Policy strategies to increase access to diabetes-appropriate foods may reduce socioeconomic inequalities in glycemic control.

Background: Food insecurity can increase risks of health and nutritional problems, leading to difficulties in self-care and poor glycemic control in diabetic patients. Purpose: This study assessed food insecurity and its association with diabetes control and self-care in type 2 diabetes patients.Methods: In this cross sectional study, 148 adults with type 2 diabetes participated. Food insecurity and self-care were determined using the 18-item USDA household food security status questionnaire and self-care (SDSCA) questionnaire. Data were analyzed using logistic and linear regression tests (SPSS 20 software).Results: Thirty-seven percent of the participants were food-insecure and significantly more likely than food-secure participants to have poor glycemic control (OR=3.02; CI: 1.45-2.65). No significant association was found between food-insecurity and overall self-care score.Conclusion: Food-insecurity was directly associated with poor glycemic control in type 2 diabetes patients. Since economic status was significantly better in the food-secure group, it can be postulated that financial problems will eventually lead to poor glycemic control.Policy strategies to increaseaccess to diabetes-appropriate foods mayreduce socioeconomic inequalities in glycemiccontrol.

Objectives Disruption of protein synthesis, by drug-mediated restriction of the ribosomal nascent peptide exit tunnel (NPET), may inhibit bacterial growth. Here, we have studied the secondary and tertiary structures of domain V of the 23S rRNA in the wild-type and mutant (resistant) H. pylori strains and their mechanisms of interaction with clarithromycin (CLA). Methods H pylori strains, isolated from cultured gastric biopsies, underwent CLA susceptibility testing by E test, followed by PCR amplification and sequencing of domain V of 23S rRNA. The homology model of this domain in H pylori, in complex with L4 and L22 accessory proteins, was determined based on the E. coli ribosome 3D structure. The interactions between CLA and 23S rRNA complex were determined by molecular docking studies. Results Of the 70 H pylori strains, isolated from 200 dyspeptic patients, 11 (16%) were CLA-resistant. DNA sequencing identified categories with no (A), A2142G (B), and A2143G (C) mutations. Docking studies of our homology model of 23S rRNA complex with CLA showed deviated positions for categories B and C, in reference to category A, with 12.19 A and 7.92 A RMSD values, respectively. In both mutant categories, CLA lost its interactions at positions 2142 and 2587 and gained two new bonds with the L4 accessory protein. Conclusion Our data suggest that, in mutant H pylori strains, once the nucleotides at positions 2142 and 2587 are detached from the drug, CLA interacts with and is peeled back by the L4 accessory protein, removing the drug-imposed spatial restriction of the NPET.

Introduction Helicobacter pylori express a large array of antigens, each of which is duly responsible for successful colonization and pathogenesis. Here, we have studied host serum antibody responses to four of its immunodominant antigens in association with the infection status and the resulting clinical outcomes. Methods For this purpose, four individual H. pylori proteins (UreB, CagA, Tip-α and HP0175) were produced in recombinant forms. Serum antibody responses of 246 (75 GC and 171 NUD) patients, against the above antigens, were evaluated by multiplex immunoblotting. The associations between the resulting data and the infection status, as well as clinical outcomes were evaluated using logistic regression models. Results Serum antibodies to all four recombinant antigens increased the chances of detecting screening ELISA-positive subjects, in an escalating dose-dependent manner, ranging from 2.6 (1.5–4.7) for HP0175 to 14.3 for UreB (4.3–50.7), exhibiting the lowest and highest odds ratios, respectively (PAdj ≤ 0.001), such that 98.2% of the subjects with antibodies to all four antigens, were also positive by the screening ELISA (P Conclusion The non-invasive multiplex serology assay, presented here, was able to not only detect subjects with current H. pylori infection, it could also screen dyspeptic patients for the presence of gastric atrophy. This simple and cost-efficient method can supplement routine screening ELISAs, to increase the chances of detecting current infections as well as atrophic gastritis.

Experiments of single-drop sedimentation and mass transfer on a laboratory scale for the extraction of acetone from a water + acetone mixture by means of the ionic liquid 1-hexyl-3-methylimidazolium hexafluorophosphate (HMIMPF6) were performed at a temperature of 298.2 K and ambient pressure. Study of the system equilibrium conditions in a previous work (Saien et al., 2015) showed that HMIMPF6, as a green solvent for the extraction of acetone from aqueous solutions, has suitable capability. The single-drop experiments with this system show that the mass transfer is not as slow as expected despite the relatively high viscosity of the system. The Henschke model is applied to the obtained data for the challenging system involving an ionic liquid by determining appropriate values for the corresponding material-specific model parameters. This model describes the experimental data with very good accuracy.

In the monster novel Gravity's Rainbow (1973), one of Thomas Pynchon's many disappearing characters prophetically announces: ‘somewhere, among the wastes of the World, is the key that will bring us back, restore us to our Earth and to our freedom’ (p. 394). Then, if anything it is ‘the wasted’, ‘the passed over’, and ‘the preterite’ that hold Pynchon's scattered narrative together. This essay focuses on the devastating proliferation of waste, the rise of technology in human affairs, and their relationship to the idea of salvation in Pynchon's work. To reflect on this link, it investigates the Calvinist rhetoric of preterition (nonelection to salvation) and its influence on contemporary US culture through Pynchon's work. First, it studies Calvin's concept of preterition and how it becomes connected to the wasted (disregarded, passed over, or used up); and second, based on Calvin's conception of preterition, it goes on to explore how the proliferation of waste in the contemporary America portrayed by Pynchon is the outcome of the capitalist economic system and its reliance on technology that converges with secularization and modernity.

Background Gastric cancer arises, mainly, on an inflammatory background. Helicobacter pylori neutrophil activating (HP-NAP) protein functions as a potent pro-inflammatory mediator. Similarly, IL-4 plays a critical role in the inflammation pathway, the levels of which are altered by C to T transition at position -590 in its promoter region. Here, we have aimed to assess the risk of gastritis and gastric cancer in the co-presence of these two inflammation modulating mediators. Methods Gastritis (n=58) and gastric cancer (n=31) patients were evaluated and compared with H. pylori-positive asymptomatic controls (n=46), for serum antibodies against recombinant HP-NAP and IL-4 C-590T single nucleotide polymorphism using immunoblotting and PCR-RFLP, respectively. Multivariable logistic regression, adjusting for age, gender and ethnicity, was used for data analysis. Results In terms of susceptibility to gastritis, seropositivity to HP-NAP projected a risk impact of 4.62 fold (OR=4.62, 95% CI=1.50-14.22), which when present in IL-4 -590 T carriers augmented the risk up to 9.7 fold (OR=9.70, 95% CI=2.06-45.69). A similar pattern, but of a stronger magnitude, occurred for the risk of gastric cancer, which was estimated at 9.07 fold (OR=9.07, 95% CI=1.99-42.0) for HP-NAP-seropositive subjects and was drastically amplified (OR=33.64, 95% CI=2.06-548.68), when double-positive (HP-NAP seropositive/IL-4 -590 T carrier) subjects were examined against double negatives (HP-NAP seronegative/IL-4 -590 CC). Conclusion Our preliminary data indicate that serum antibodies against HP-NAP represent a state of risk, which is further exacerbated in IL-4 -590 T carriers. These biomarkers, if validated in larger prospective studies, can be used to screen for gastric cancer susceptibility.

BACKGROUND Fluoroquinolones hinder bacterial DNA replication by inhibiting DNA gyrase. However, mutations, in the QRDR segment of its A subunit (GyrA), cause antibiotic resistance. Here, the interactions of levofloxacin (LVX), gemifloxacin (GXN), and moxifloxacin (MXN) with Helicobacter pylori GyrA, in LVX-resistant vs -sensitive strains, were studied. METHODS Levoflixacin-sensitive (n = 4) and -resistant (n = 9) H pylori strains, randomly selected from another antibiotic susceptibility study, underwent PCR amplification of gyrA gene, spanning the QRDR segment. The amplified gene fragments were sequenced and aligned. The homology model of H pylori GyrA was built based on that of Escherichia coli, and energy minimization was done. The interaction patterns of LVX, GXN, and MXN with GyrA were analyzed via molecular docking studies. RESULTS Sequence alignment of the 13 studied strains, created 5 categories of strains: (A) wild type-like (H pylori ATCC26695), (B) N87K-only, (C) D91N-only, (D) N87K + V94L, and (E) D91N + A97V mutations. The minimum inhibitory concentrations (MIC) for LVX-sensitive (category A) and -resistant (categories B-E) strains were

Background: Two of the Wnt signaling pathway target genes, tumor necrosis factor receptor family member (TROY) and leucine-rich G-protein coupled receptor (LGR5), are involved in the generation and maintenance of gastrointestinal epithelium. A negative modulatory role has recently been assigned to TROY, in this pathway. Here, we have examined their simultaneous expression in gastric carcinogenesis. Methods: Tumor and paired adjacent tissues of intestinal-type gastric cancer (GC) patients (n = 30) were evaluated for LGR5 and TROY expression by immunohistochemistry. The combination of the percentage of positively stained cells and the intensity of staining was defined as the composite score and compared between groups. The obtained findings were re-evaluated in a mouse model. Results: TROY expression in the tumor tissue was significantly lower than that of the adjacent tissue (2.5 ± 0.9 vs. 3.3 ± 0.9, p = 0.004), which was coincident with higher LGR5 expression (3.6 ± 1.1 vs. 2.7 ± 0.9, p = 0.001). This observation was prominent at stages II/III of GC, leading to a statistically significant mean difference of expression between these two molecules (p = 0.005). In the H. pylori infected-mouse model, this inverse expression was observed in transition from early (8-16 w) to late (26-50 w) time points, post treatment (p = 0.002). Conclusion: Our data demonstrates an inverse trend between TROY down-regulation and LGR5 up-regulation in GC tumors, as well as in response to H. pylori infection in mice. These findings support a potential negative modulatory role for TROY on LGR5 expression.

The influence of naturally occurring monovalent NaCl and divalent Na 2 SO 4 salts with anions at near extremes of Hafmester series on liquid–liquid equilibrium (LLE) of the chemical system of {water + acetone + 1-Hexyl-3-methylimidazolium hexafluorophosphate (HMIMPF 6 ) ionic liquid} was studied. The conditions were 298.2 K, atmospheric pressure of 81.5 kPa and salts mass fractions in initial aqueous solutions of 0.05 and 0.10. Results show that salting-out effect was significant, so that enhancements in the acetone distribution coefficient was achieved within (21.67–35.54)% for NaCl and (51.84–92.37)% for Na 2 SO 4 . The stronger anion activity and the lack of anion exchange with the used ionic liquid can be the reasons for the stronger effect of the latter salt. The consistency of the experimental data were examined with the improved Eisen-Joffe equation. For modeling purpose, the NRTL and UNIQUAC equations were satisfactorily applied to correlate the phase equilibria. In the presence of NaCl and Na 2 SO 4 salts, the root mean square deviations of compositions, for NRTL equation were 0.0346 and 0.0634 and for UNIQUAC equation 0.1093 and 0.0328, respectively.

BabA is a Helicobacter pylori cell surface adhesin, which binds to the ABO/Le(b) histo-blood group antigens (Le(b)) and serves as a virulence factor.H. pylori single colonies were isolated from 156 [non-ulcer dyspepsia (NUD) = 97, duodenal ulcer (DU) = 34, gastric cancer (GC) = 25)] patients. babA and babB genes were evaluated by gene/locus-specific PCR. BabA protein expression and Le(b) binding activity were determined by immunoblotting and ELISA, respectively.The combined categorization of H. pylori strains based on high, low or no levels of BabA expression and Le(b) binding, produced 4 groups: (I) BabA-high/Le(b)-high (36 %), (II) BabA-low/Le(b)-low (26 %), (III) BabA-neg/Le(b)-low (30 %) and (IV) BabA-neg/Le(b)-neg (8 %) strains. The majority (63 %) of the BabA-low/Le(b)-low strains exhibited mixed babA/B genotypes as compared to merely 18 % of the BabA-high/Le(b)-high, 15 % of the BabA-neg/Le(b)-neg and 11 % of the BabA-neg/Le(b)-low (P = 0.0001) strains. In contrast to NUD strains, the great majority (70 %) of DU strains were BabA-low/Le(b)-low (11 %, P = 0.0001), which compared to NUD strains, enhanced the risk of DU by 18.8-fold. In parallel, infection with babA/B mixed genotype strains amplified the risk of DU by 3.6-fold (vs. babA-positive: P = 0.01) to 6.9-fold (vs. babA-negative: P = 0.007).Here, we show higher prevalence of mixed babA/B genotypes among BabA-low/Le(b)-low clinical strains. Recombination of babA and babB genes across their loci may yield lower BabA expression and lower Le(b) binding activity. We conclude that H. pylori strains with lower Le(b) binding activity are better adapted for colonization of the gastric metaplastic patches in the duodenum and enhance the risk of duodenal ulcers.

Gastric cancer is the fourth most prevalent cancer worldwide, which claims more than 700,000 lives every year. However, the current chemical and surgical management of this mortal cancer remains far from successful. A recent safe approach to preventing or treating tumorigenesis has been the re-introduction of traditional herbal medicine, taking advantage of efficient, yet natural healthy products. In this review, we have focused on a number of such popular agents, mostly introduced by the Traditional Chinese medicine (TCM), which have exhibited highly significant preventive/therapeutic effects against various gastric pathologies, ranging from gastritis to premalignant lesions and confirmed malignancies, in experimental rats. The underlying mechanisms of action seem to concentrate on shifting the balance between in vivo oxidant and anti-oxidant activities. In addition, the existing reports on the potential toxicity of these compounds, as well as their interactions with chemical drugs, have been presented. However, far more details are required to recommend herbal medicine as an effective and reliable plan of action in gastric cancer management.

(Liquid + liquid) equilibrium (LLE) of the chemical system of {water + acetone + 1-Hexyl-3-methylimidazolium hexafluorophosphate (HMIMPF6) ionic liquid} was studied at different temperatures of (293.2, 298.2 and 303.2) K and under atmospheric pressure of 81.5 kPa. The results show that HMIMPF6 provides the acetone distribution coefficient and separation factor values within (0.8813 to 1.2351) and (3.0 to 54.4), respectively; indicating the high capability of the ionic liquid for extraction of acetone from aqueous solutions. In most cases, acetone solubility in the ionic liquid is higher than in water, especially at higher solute concentrations. Meanwhile, higher separation factor is relevant to the lower temperature due to lower (water + ionic liquid) miscibility. The consistency of tie line data, at each temperature, was examined with Othmer–Tobias correlation. The values were nicely reproduced with the well-known NRTL and UNIQUAC models. Accordingly, the required thermodynamic properties of HMIMPF6 were obtained by the Density Functional Theory (DFT) calculations, carried out at the M06/6-311++G∗∗ level of theory. The root mean square deviations (RMSD) between experimental and model concentration values were 0.0192 and 0.0255, respectively; indicating close agreement of the both models.

Background Most two- dimensional in vitro models for studying host- H. pylori interactions rely on tumor-derived cell lines, which harbor malignant alterations. The recent development of human gastric organoids has overcome this limitation and provides a highly sophisticated, yet costly, short-term model for H. pylori infection, with restricted use in low-budget centers. Method Tissue specimens from upper, middle, and lower stomachs of H. pylori-negative volunteers were collectively dispersed and cultured on mouse embryonic fibroblast (MEF) or collagen-coated plates. Gastric primary cells (GPCs) were evaluated by light microscopy, immunostaining, qRT-PCR and ELISA analysis of cellular secretions, before and after H. pylori infection. Results The formation and long-term (up to 1 year) maintenance of GPCs was highly dependent on adherent inactivated MEF cells, cultured in enriched media. These cells were multipassageable and able to undergo stable freezer storage and subsequent revival. The cellular composition of GPCs included the combination of cytokeratin 18 (CK18) and E-cadherin (E-cad)-positive epithelial cells, MUC5AC-positive gastric cells, and leucine-rich repeat containing G protein-coupled receptor 5 (LGR5)-positive progenitor cells. These cells produced significant amounts of gastric pepsinogens I and II. GPCs also allowed for extended (up to 96 hours) H. pylori infection, during which they underwent morphological alterations (cellular vacuolation and elongation) and hyperproduction of gastric pepsinogens and inflammatory cytokines (IL-8 and TNF-α). Conclusion We, hereby, present a simple, consistent, and cost-efficient gastric cell culture system, which provides a suitable model for extended in vitro infection of H. pylori. This platform can be employed for a variety of gastric-related research.

A remarkable disadvantage of pure ionic liquids, to be used as a solvent in different processes, is their high viscosity and density. Here, n-butyl acetate was used as a cosolvent in the liquid–liquid system of {water + acetone + 1-hexyl-3-methylimidazolium hexafluorophosphate (HMIMPF6) ionic liquid}. Mixtures of the ionic liquid and the cosolvent were used as the organic phase to extract acetone from aqueous phase. A drastic decrease in viscosity and a significant decrease in density of the ionic liquid were achieved in the presence of cosolvent. Accordingly, the liquid–liquid equilibrium of different provided systems was studied under temperature of 298.2 K and atmospheric pressure of 81.5 kPa. Binary phase regions and corresponding tie-lines and binodal curves were obtained for ternary and quaternary systems. Results show the extension of the binodal solubility region as well as enhancements up to 41.8% in the acetone separation factor. The tie-line data consistency was confirmed by the Othmer–Tobias and the Hand equations. Moreover, the nonrandom two-liquid model of activity coefficient was applied satisfactorily to reproduce the equilibrium data with a root-mean-square deviation of only 1.95%.

Treatment success of H. pylori eradication therapy has declined worldwide largely because of increased antimicrobial resistance. New therapeutic approaches are needed, especially for countries like Iran, where resistance to commonly used drugs is already widespread and traditional H. pylori therapies produce poor cure rates. To review the results of quadruple therapy trials containing bismuth and furazolidone in Iran. We searched PubMed, Google scholar as well as the references of all published papers for studies conducted in Iran, utilizing furazolidone in the treatment of H. pylori infections. The target population was four drug studies that utilized a combination of bismuth, furazolidone, amoxicillin, or tetracycline plus a proton pump inhibitor. Eighteen studies with 22 arms including 1713 subjects were found. The weighted mean cure rate for 14-day studies (six studies) using 200 mg b.i.d. furazolidone was 80% intention to treat (ITT) and 87% per protocol (PP). Studies using 100 mg b.i.d. (three studies) were less effective (weighted mean ITT cure rate = 67%). One small 14-day study with furazolidone 100 mg q.i.d. achieved cure rates of 94.5% ITT and PP. Although furazolidone–bismuth quadruple therapy proved relatively effective in Iran, furazolidone-containing regimens remain to be optimized. Based on these data and results from China, it appears likely that 14-day therapy containing furazolidone 100 mg t.i.d. or q.i.d. is likely to provide the highest cure rates with lowest side effects; this remains to be experimentally tested. Detailed suggestions for further development of furazolidone-containing regimens are provided.

International audience; Helicobacter pylori, a microaerophilic fastidious bacterium, has been cultured on various plating and broth media since its discovery. Although the agar media can be sufficient for the identification, typing, and antibiotic resistance studies, no secretory antigen of H. pylori can be evaluated in such media. Thus, satisfactory growth of H. pylori in liquid culture which is needed for analysis of secretory proteins without the presence of interfering agents is in demand. We assessed the impact of beta-cyclodextrin, Fetal Bovine Serum (FBS), and charcoal as supplements for H. pylori growth. Furthermore, we aimed to identify the most favorable supplement that supports the secretion of the dominant secretory protein, vacuolating cytotoxin (VacA). Five clinical strains were cultured on broth media and the growth, viability, morphology, and protein content of each strain were determined. Our results revealed that beta-cyclodextrin supports the growth rate, viability, and cell lysate protein content to the extent similar to FBS. Application of beta-cyclodextrin is found to postpone spiral to coccoid conversion up to 72 h of incubation. Although FBS supports a higher VacA protein content, presence of interfering macromolecules in FBS questions its utility particularly for purposes of studying extra cellular proteins such as VacA. This study recommends further application of beta-cyclodextrin as a culture supplement with the potential capacity in neutralizing toxic compounds and flourishing the secretion of H. pylori proteins without addition of interfering proteins.

Helicobacter pylori secretory peptidyl prolyl isomerase, HP0175, is progressively identified as a pro-inflammatory and pro-carcinogenic protein, which serves to link H. pylori infection to its more severe clinical outcomes. Here, we have analyzed host HP0175-specific antibody responses in relation to the severity of gastritis. The HP0175 gene fragment was PCR-amplified, cloned, expressed and purified by Ni-NTA affinity chromatography. Serum antigen-specific antibody responses of non-ulcer dyspeptic patients (N = 176) against recombinant HP0175 were detected by western blotting. The infection status of these subjects was determined by rapid urease test, culture, histology, and serology. The grade of inflammation and stage of atrophy were scored blindly according to the OLGA staging system. The recombinant HP0175 (rHP0175) was expressed as a ~35 kDa protein and its identity was confirmed by western blotting using anti-6X His tag antibody and pooled H. pylori-positive sera. Serum IgG antibodies against rHP0175 segregated our patients into two similar-sized groups of sero-positives (90/176, 51.1 %) and sero-negatives (86/176, 48.9 %). The former presented with higher grades of gastric inflammation (OR = 4.4, 95 % CI = 1.9–9.9, P = 0.001) and stages of gastric atrophy (OR = 18.3, 95 %CI = 1.4-246.6, P = 0.028). Our findings lend further support to the pro-inflammatory nature of H. pylori peptidyl prolyl isomerase (HP0175) and recommends this antigen as a non-invasive serum biomarker of the severity of H. pylori-associated gastritis.

Helicobacter pylori (H. pylori) is a common infection responsible for considerable morbidity and mortality worldwide. It is now recommended that all H. pylori infections be cured unless there are compelling reasons not to. Like other infectious diseases, the most reliable method to achieve high cure rates is to base therapy on the results of patient-specific susceptibility testing. Empiric therapy must be based on knowledge of the local susceptibility patterns tempered by data obtained from the patient concerning their antibiotic exposure. Data from clinical trials without susceptibility testing cannot be reliably used in other locations. Because of increasing resistance, triple therapies based on clarithromycin, metronidazole, or levofloxacin should only be used for susceptibility-based therapy. If there are no options other than to give empiric therapy, only those regimens proven to be effective locally should be used. Special efforts should be made to enhance patient adherence. In most regions 14-day four-drug combinations are required for successful empiric therapy. Testing to confirm cure should be routinely done.

Background and Aims: Helicobacter pylori is a highly diverse pathogen, which encounters epithelial cells as the initial defense barrier during its lifelong infection. The structure of epithelial cells can be disrupted through cleavage of microfilaments. Cytokeratin 18 (CK18) is an intermediate filament, the cleavage of which is considered an early event during apoptosis following activation of effector caspases. Methods: Helicobacter pylori strains were isolated from 76 dyspeptic patients. cagA 3’ variable region and CagA protein status were analyzed by PCR and western blotting, respectively. Eight hours post-co-culture of AGS cells with different H. pylori strains, flow cytometric analysis was performed using M30 monoclonal antibody specific to CK18 cleavage-induced neo-epitope. Results: Higher rates of CK18 cleavage were detected during co-culture of AGS cells with H. pylori strains bearing greater numbers of cagA EPIYA-C and multimerization (CM) motifs. On the other hand, H. pylori strains with greater numbers of EPIYA-B relative to EPIYA-C demonstrated a decrease in CK18 cleavage rate. Thus, H. pylori-mediated cleavage of CK18 appeared proportional to the number of CagA EPIYA-C and CM motifs, which seemed to be downplayed in the presence of EPIYA-B motifs. Conclusions: Our observation associating the heterogeneity of cagA variants with the potential of H. pylori strains in the induction of CK18 cleavage as an early indication of apoptosis in gastric epithelial cells supports the fact that apoptosis may be a type-specific trait. However, additional cagA-targeted experiments are required to clearly identify the role of EPIYA and CM motifs in apoptosis and/or the responsible effector molecules.

Background: Helicobacter pylori (Hp) is a bacterium recognised as a main causative agent for the development of chronic active gastritis, peptic ulcer disease, gastric adenocarcinoma and primary gastric lymphoma. Objective: Determination of the levels of IFN-γ (pro-inflammatory) and IL-4 (anti inflammatory) cytokine expression as indicators of Th1 and Th2 immune responses in gastric cancer (GC) and non gastric cancer (Non GC) dyspeptic patients by gene specific RT-PCR. Materials and Methods: Biopsy specimens were collected from three groups of gastric cancer (GC=18), non ulcer dyspepsia (NUD = 38) and peptic ulcer patients (PUD=20). Total RNA was extracted and complementary DNA was synthesised. PCR amplification was performed for HPRT, IFN-γ and IL-4 cytokines and the intensity of each band was measured by densitometry and normalized against HPRT expression as a house keeping gene. Results: Comparison of the results from different groups of patients indicated that IFN-γ gene expression was similar in nonGC dyspeptic patients (NUD and PUD groups; 3.38 ± 0.57,3.43 ± 0.41, respectively) whereas, in GC patients, it was significantly higher than others (5.52 ± 0.59; P < 0.0001). On the other hand, IL-4 gene expression showed no significant difference between NUD and GC patients (2.81 ± 0.43,2.3 ± 0.12 respectively), whereas the expression rate of this cytokine was significantly higher in PUD patients (3.7 ± 0.1; P 0.05). Our data indicate an association between Th1 and Th2 immune responses and the development of gastric cancer and peptic ulcer disease respectively.

The definite mode of transmission of Helicobacter infection is largely unknown. This study was carried out primarily, to determine the existence of Helicobacter spp. in the oral secretions of stray cats as one of the possible routes of transmission and secondly, to evaluate the accordance between oral and gastric colonization of Helicobacter spp. in these cats. Forty-three adult stray cats were thus studied for the presence of Helicobacter species by quantitative rapid urease test (RUT), cytology and PCR. Helicobacter spp. were found in the oral secretions and gastric biopsies of 93% and 67.5% of the stray cats, respectively. There was not, however, any agreement observed between Helicobacter colonization at these two locations, at neither genus nor species level. These findings suggest that the oral cavity is routinely exposed to transient forms of bacteria and may temporarily harbor Helicobacter spp. Thus, oral cavity as a source of Helicobacter spp. may act as a reservoir for transmission and may not necessarily reflect the colonization status of the gastric mucosa.

Helicobacter-like organisms are frequently found in canine stomachs, but the relationship between such organisms and gastric pathology has not been established. However, some such organisms have zoonotic importance. The aims of this study were to evaluate the morphological and biochemical characteristics of cultivable canine gastric Helicobacter-like organisms (GHLOs) in pets and stray dogs and their prevalence in these two groups of dogs. Specimens were taken by gastroscopy from 30 clinically healthy stray dogs and 30 pet dogs. Cultures were positive from biopsies of 11/30 of stray and 6/30 of pet dogs. The isolated Helicobacters were observed by light microscopy and studied by biochemical, physiological and PCR analysis. Some of the isolated GHLO's displayed atypical shapes that were similar to Helicobacter pylori or Helicobacter acinonychis in stray dogs' cytological examinations. They had 2-3 helices and were smaller than other canine GHLOs. One of these atypical Helicobacter strains was cultured. It was not possible to distinguish such strains by routine PCR and biochemical evaluations. Electron microscopy showed a smaller Helicobacter (2 microm in length) with 2 or 3 helixes. This study demonstrates that not all canine gastric Helicobacters are 5-15 microm in length, as has been previously proposed, and portrays the need for further investigation of canine GHLOs.

Background/Aim: The identification of the vacA intermediate region has provided new insights into the role of vacA heterogeneity in relation to gastro-duodenal pathogenesis. The aim of this study was to assess vacA polymorphism in Iranian Helicobacter pylori strains and its association with cagA as a major virulence determinant, gastric histopathology and disease. Methods:vacA polymorphism and serum antibody responses were studied in 207 H. pylori-infected (139 NUD, 34 PUD, and 34 GC) patients and correlated with gastric histopathology. Results: Multivariate logistic regression analysis found intermediate region typing superior to signal or mid region typing for screening high risk patients. vacA i1 allele was identified as an independent predictor of dysplasia (OR = 9.044; 95% CI: 1.11–73.33). Possession of s1/i1/cagA+ strains was also identified as a predictor of intestinal metaplasia (OR = 3; 95% CI: 1.13–7.95), dysplasia (OR = 9.9; 95% CI: 1.23–80.86) and risk of GC (OR = 6.9; 95% CI: 2.5–18.66) as well as induction of anti-VacA sero-positivity (OR = 5.04; 95% CI: 1.8–13.6). Anti-VacA serology correctly detected 83.8% of s1/i1/cagA+ strains carried by high-risk patients. Conclusions: The current study emphasizes the implication of vacA polymorphic structure, especially the s1/i1/cagA+ genotype, in increasing the risk of GC by revealing their association with gastric pre-neoplastic changes and their reflection in VacA sero-positivity which encourages the application of noninvasive procedures in population screening.

The aim of this study was to evaluate the prevalence of chronic gastritis in pet dogs, to determine the histopathologic changes of gastric mucosa and, to determine its relationship with canine gastric Helicobacter infection. Sixty percent (n = 18), 27% (n = 8) and 13% (n = 4) of the examined stomachs showed normal, congested and erosive gastric mucosa respectively. Histopathologic examination was confirmed the presence of chronic gastritis in 40% of dogs (n = 12). Lymphocytic-plasmacytic gastritis was the most common type of chronic gastritis. Gastric Helicobacter was detected in cytological examination of 26 out of 30 dogs (86.6%) but in the PCR analysis, 93% of gastric samples were positive for GHLO. There was no significant relation between the presence of Helicobacters and chronic gastritis (p>0.05). Follicular gastritis was detected in 12 cases (40%) and there was also no significant correlation between its presence and GHLO's infection (p>0.05). In conclusion, chronic gastritis can be considered as a prevalent disease especially in dogs. Nutritional and environmental factors as well as individual immune response may have role in induction of chronic gastritis, but the clinical significance of these histopathologic changes should be evaluated.