Helicobacter pylori infection is a major driver of gastric cancer, with the CagA oncoprotein promoting chronic inflammation and epithelial injury. However, the overall picture of the cytokine–receptor pathways involved in this process remains unclear. To investigate how CagA modulates epithelial signaling, we generated cagA-deletion and cagA-complemented mutants of the gerbil-adapted TN2 strain and infected AGS gastric epithelial cells for 6–48 h. We also analyzed 85 gastric biopsy specimens from a nationwide Indonesian survey. In vitro, wild-type and cagA-complemented H. pylori robustly induced IL-8, IL-1β, and TNF-α, as expected. In the CagA-positive strain, the newly identified anti-inflammatory cytokine, IL-13, was also induced. IL-13-related Fn14, a TNF receptor superfamily member, was strongly upregulated in a CagA-positive strain, whereas TNFR1 remained stable, and TNFR2 was consistently downregulated. In clinical specimens, Fn14 expression was elevated in atrophic gastritis and closely associated with cagA positivity. IL-13 expression showed a distinct pattern, increased in atrophic gastritis but unrelated to H. pylori or cagA status, instead correlating positively with TWEAK, the ligand for Fn14. Together, these findings suggest that cagA-positive H. pylori infection is associated with Fn14 upregulation, whereas H. pylori infection in general is linked to TNFR2 downregulation. IL-13-related signaling may instead reflect mucosal injury and repair responses, indicating context-dependent regulation of TNF receptor family pathways in gastric mucosal inflammation.

BACKGROUND:Variations in Helicobacter pylori infection rates and pathogenicity do not explain the global gastric cancer incidence, indicating that other bacteria may play a role. We investigated the pathogenic factors of H. pylori and their interactions with the gastric microbiome in a population with low gastric cancer but high gastritis rates in Indonesia. METHODS:The study included 66 H. pylori-positive gastric biopsies. DNA was extracted from the bacterial cultures to examine the pathogenic factors of H. pylori. The 16S rRNA V3-V4 region was sequenced using next-generation sequencing. The microbiome analysis concentrated on α-diversity and β-diversity, along with absolute and relative abundances. Correlation analysis and predicted functional inference were conducted using SECOM and PICRUSt2. RESULTS:Helicobacter predominates in H. pylori-infected stomachs, limiting other bacteria. Although α-diversity was non-significant, virulent H. pylori genotypes showed greater microbial diversity, suggesting co-colonization by other taxa. Some taxa were notably abundant across pathogenic subtypes (p < 0.05), such as Veillonella sp. in East Asian-type CagA H. pylori and Klebsiella without babB. The β-diversity results indicated that microbial diversity and abundance varied according to polymorphisms in patients with different H. pylori CagA types, sabA status, homA/B, and iceA subtypes (PERMANOVA test; p < 0.05). H. pylori dominance remains unchanged when atrophy worsens, alongside decreased microbial diversity (p < 0.05 for atrophy stage 0 vs. stages 1 and 2). Microbial correlation analysis revealed that Helicobacter only had a positive linear relationship with Veillonella (SECOM(Pearson2) = 0.51, SECOM(Distance) = 0.60), whereas Streptococcus sp. correlated with several gastric taxa. Predicted functional inference showed several pathways to be depleted when atrophy progresses. CONCLUSION:Various pathogenic factors impact microbial diversity, and bacteria cohabiting in the gastric environment might shape disease outcomes. Additionally, our study uncovers relationships among genera present in the stomach. More research is needed to explore how non-Helicobacter species induce or possibly safeguard against gastric pathologies.

BACKGROUND:Despite the low prevalence of Helicobacter pylori in Indonesia, the high incidence of gastritis, predominantly atrophic gastritis, suggests that factors such as autoimmune gastritis (AIG) contribute to this unusual pattern. This study aims to investigate the epidemiology of AIG, histopathology, and its association with H. pylori status in Indonesia. METHODS:A cross-sectional study was conducted in various regions in Indonesia between 2014 and 2017; 380 eligible sera and gastric biopsies were available when this study was conducted. As many as 138 sera samples were included in this study based on the initial examination by the updated Sydney system. The diagnosis of AIG was confirmed by serologic testing for parietal-cell antibodies (PCA) and detailed histopathological assessment with sparing of antrum histopathological features. RESULTS:Among the included samples in this study, 78.99% (109/138) were PCA positive (≥ 10 RU/mL) and 0.72% (1/138) were considered to be diagnosed as AIG (spared from antrum histopathological features). The majority of PCA positive cases were H. pylori positive (61/109; 55.96%) with a significant correlation (p < 0.05, R = 0.31). Additionally, a significant association was found between H. pylori infection and PCA level with gastric histopathological features (p < 0.05). CONCLUSION:This study demonstrates that the incidence of gastritis without H. pylori infection in Indonesia is not attributable to AIG, as only a single AIG-positive case was found. These findings underscore the important role of H. pylori as a pathogenic factor in chronic gastritis and highlight its mechanisms in triggering immune responses and driving disease progression and histopathological changes.

Neoadjuvant chemotherapy (NAC) followed by surgery is a standard approach for management of locally advanced esophageal squamous cell carcinoma (ESCC). Patients who do not respond well to NAC have a poor prognosis. Despite extensive research, the mechanisms of chemoresistance in ESCC remain largely unknown. Here, we established paired tumor organoids—designated as PreNAC-O and PostNAC-O—from one ESCC patient before and after NAC, respectively. Although the two organoids did not exhibit significant differences in proliferation, morphology or drug sensitivity in vitro, the tumorigenicity of PostNAC-O in vivo was significantly higher than that of PreNAC-O. Xenografts from PreNAC-O tended to exhibit keratinization, while those from PostNAC-O displayed conspicuous necrotic areas. The tumorigenicity of PostNAC-O xenografts during the chemotherapy was comparable to that of PreNAC-O without treatment. Furthermore, the gene expression profiles of the xenografts suggested that expression of genes involved in the EMT and/or hypoxia response might be related to the tumorigenicity of PostNAC-O. Our data suggested that the tumorigenicity of residual cancer had been enhanced, outweighing the effects of chemotherapy, rather than being attributable to intrinsic chemoresistance. Further studies are required to clarify the extent to which residual cancers share a common mechanism similar to that revealed here.

GroundBIRD is a ground-based cosmic microwave background (CMB) experiment for observing the polarization pattern imprinted on large angular scales (ℓ > 6 ) from the Teide Observatory in Tenerife, Spain. Our primary scientific objective is a precise measurement of the optical depth τ (σ(τ ) ∼ 0.01) to the reionization epoch of the Universe to cross-check systematic effects in the measurements made by previous experiments. GroundBIRD observes a wide sky area in the Northern Hemisphere (∼ 40% of the full sky) while continuously rotating the telescope at a high speed of up to 20 rotations per minute (rpm) to overcome the fluctuations of atmospheric radiation. We have adopted the NbTiN/Al hybrid microwave kinetic inductance detectors (MKIDs) as focal plane detectors. We observe two frequency bands centered at 145 GHz and 220 GHz. The 145 GHz band picks up the peak frequency of the CMB spectrum. The 220 GHz band helps accurate removal of the contamination of thermal emission from the Galactic interstellar dust. The MKID arrays (138 MKIDs for 145GHz and 23 MKIDs for 220GHz) were designed and optimized so as to minimize the contamination of the two-level-system noise and maximize the sensitivity. The MKID arrays were successfully installed in May 2023 after the performance verification tests were performed at a laboratory. GroundBIRD has been upgraded to use the full MKID arrays, and scientific observations are now underway. The telescope is automated, so that all observations are performed remotely. Initial validations, including polarization response tests and observations of Jupiter and the moon, have been completed successfully. We are now running scientific observations.

Background: Helicobacter pylori (H. pylori) infection status in Indonesia might be higher than the actual prevalence. A comprehensive epidemiologic study is necessary to explore infection susceptibility by demographic characteristics, especially between Indonesia’s Eastern and Western regions. Methods: We included our recent survey and data from our previous studies with 1,172 endoscopic examinations from 19 cities throughout Indonesia from January 2014 to March 2017. H. pylori infection was diagnosed based on the combination of four different tests. Using a criterion of one positive result between four tests, we found that 190 subjects (190/1,158; 16.4%) were H. pylori positive. A total of 939 subjects were analyzed for risk factors of sociodemographic characteristics to H. pylori infection. Results: Prevalence was slightly higher in the Eastern region compared with the Western region of Indonesia (79/277; 59.4% vs. 54/529; 40.6%). Living in the Eastern region imposes a higher risk of infection (OR 5.33, 95%CI 1.17–24.26). Timor ethnicity had the highest prevalence (15/28; 53.6%), followed by Buginese ethnic (21/74; 28.4%), and Papuan ethnic (18/64; 28.1%) in the Eastern region. As an ethnicity with a high prevalence (37/109; 33.9%), Batak was an outlier among the low prevalence areas in the Western region. In general, age, living in the Eastern region, Batak ethnicity, Protestant, Catholic, and alcohol drinking were independent risk factors associated with H. pylori infection in Indonesia (OR 1.03, 95%CI 1.02–1.05; OR 5.33, 95%CI 1.17–24.26; OR 5.89, 95%CI 1.58–21.94; OR 5.47, 95%CI 2.16–13.86; OR 5.05, 95%CI 1.43–17.80; OR 2.30, 95%CI 1.14–4.61, respectively). Conclusions: Several ethnicities and habits were concluded to increase the risk of infection as the interaction of host, agent, and environment is inseparable in affecting disease susceptibility.

Background/Aims:: A latex agglutination turbidity (LA) assay to test for serum antibodies has been approved in Japan and Korea for mass screening of Helicobacter pylori infection. In this study, we evaluated the LA assay for diagnosing H. pylori infection and predicting gastric mucosal changes in a Mongolian population. Methods:: In total, 484 individuals were classified into H. pylori-positive (n=356) and H. pylori-negative (n=128) groups, as determined by histology and H. pylori culture. Results:: The best cutoff, sensitivity, and specificity values for the LA assay were 18.35 U/mL, 74.2%, and 65.6%, respectively. The LA values in the atrophic gastritis group were statistically higher than those in the other groups (healthy, chronic gastritis, intestinal metaplasia, and gastric cancer, p<0.0001). The cutoff value to distinguish the atrophic gastritis group from the other four groups was 32.0 U/mL, and its area under the curve was 0.673, which was the highest among the E-plate, pepsinogen (PG) I, PG II, and PG I/II ratio tests in our data. The odds ratios for atrophic gastritis determined by the LA assay and PG I test in multiple logistic regression were 2.5 and 1.9, respectively, which were significantly higher than for the other tests. Conclusions:: The LA assay can determine the risk of atrophic gastritis, which in turn is a considerable risk factor for gastric cancer. We propose using this assay in combination with the PG I/II ratio to avoid missing gastric cancer patients who have a low LA value (less than 32.0 U/mL).

INTRODUCTION:We have previously reported that overexpression of visinin-like protein 1 (VSNL1) is frequently observed in advanced colorectal adenocarcinomas and correlates with poorer prognosis. In this study, we determined the levels of VSNL1 expression in the earlier stages of colorectal tumors including adenomas and adenocarcinomas, and attempted to clarify the functional significance of VSNL1 overexpression in colorectal carcinogenesis.METHODS:Levels of VSNL expression in colorectal tumor tissues were analyzed using immunohistochemistry. The effects of VSNL1 downregulation and overexpression on cell proliferation, resistance to apoptosis, and invasiveness were determined using two VSNL1-overexpressing colorectal cancer cell lines, CW-2 and HCT-116 and VSNL1 inducibly expressing SNU-C5, respectively. Gene expression signatures in VSNL1-downregulated CW-2 and HCT-116 were identified using transcriptome and gene set enrichment analyses.RESULTS:VSNL1 expression was restricted to only a few crypt cells in the non-tumorous epithelium, whereas it became enhanced in adenomas and adenocarcinomas with the progression of tumorigenesis. Downregulation of VSNL1 in CW-2 and HCT-116 cells suppressed their proliferation through induction of apoptosis. Conversely, overexpression of VSNL1 in SNU-C5 cells enhanced resistance to anoikis. Transcriptome and gene set enrichment analyses revealed that downregulation of VSNL1 altered the expression level of the apoptosis-related gene set in CW-2 and HCT-116 cells.CONCLUSION:VSNL1 plays a role in both the development and progression of colorectal tumors by enhancing cell viability.

Colorectal cancer (CRC) is one of the most frequent types of cancer, with high incidence rates and mortality globally. The extended timeframe for developing CRC allows for the potential screening and early identification of the disease. Furthermore, studies have shown that survival rates for patients with cancer are increased when diagnoses are made at earlier stages. Recent research suggests that the development of CRC, including its precancerous lesion, is influenced not only by genetic factors but also by epigenetic variables. Studies suggest epigenetics plays a significant role in cancer development, particularly CRC. While this approach is still in its early stages and faces challenges due to the variability of CRC, it shows promise as a potential method for understanding and addressing the disease. This review examined the current evidence supporting genetic and epigenetic biomarkers for screening and diagnosis. In addition, we also discussed the feasibility of translating these methodologies into clinical settings. Several markers show promising potential, including the methylation of vimentin (VIM), syndecan-2 (SDC2), and septin 9 (SEPT9). However, their application as screening and diagnostic tools, particularly for early-stage CRC, has not been fully optimized, and their effectiveness needs validation in large, multi-center patient populations. Extensive trials and further investigation are required to translate genetic and epigenetic biomarkers into practical clinical use. biomarkers, diagnostic biomarkers.

Patient-derived tumor organoids have considerable potential as an in vitro diagnostic tool for drug susceptibility testing. In the present study, we investigated whether bile collected for diagnostic purposes could be a potential source for the establishment of biliary cancer organoids. Among 68 cases of biliary cancer, we successfully generated 60 bile-derived organoids (BDOs) from individual patients. Consistent with previous reports that described biliary cancer organoids from surgical tissues, the BDOs showed diverse morphologies such as simple cysts, multiloculated cysts, thick capsulated cysts, and solid masses. They also harbored mutations in KRAS and TP53 at frequencies of 15% and 55%, respectively. To enrich the cancer organoids by removing contaminated noncancerous components of BDOs, we attempted to verify the effectiveness of 3 different pro-cedures, including repeat passage, xenografting, and selection with an MDM2 inhibitor for TP53 mutation-harboring BDOs. By monitoring the sequence and expression of mutated TP53, we found that all these procedures successfully enriched the cancer organoids. Our data suggest that BDOs can be established with minimal invasiveness from almost all patients with biliary cancers, including inoperable cases. Thus, despite some limitations with respect to the characterization of BDOs and methods for the enrichment of cancer cell-derived organoids, our data suggest that BDOs have potential applications in personalized medicine.(c) 2023 THE AUTHORS. Published by Elsevier Inc. on behalf of the United States & Canadian Academy of Pathology. This is an open access article under the CC BY license (http:// creativecommons.org/licenses/by/4.0/).

Neoadjuvant chemotherapy (NAC) followed by surgery is one of the standard therapeutic approaches in Japan for patients with locally advanced esophageal carcinoma. Recently, the JCOG1109 study revealed that NAC with docetaxel, cisplatin and 5-fluorouracil (5-FU) (DCF-NAC) is superior to NAC with cisplatin and 5-FU, and has now become the standard preoperative chemotherapy. Using a microarray system, we have previously investigated the expression profiles of endoscopic biopsy samples from patients with esophageal squamous cell carcinoma (ESCC) before DCF-NAC (preNAC) and identified 17 molecules as biomarkers predictive of a pathologically complete response to DCF-NAC. Here, we re-grouped our previous dataset based on the histopathological response grade with the addition of several microarray profiles and conducted a re-analysis using bioinformatic web tools including DAVID, GSEA, UALCAN, and CIBERSORTx. We identified 204 genes that were differentially expressed between the highly resistant and sensitive groups. Some of these differentially expressed genes (DEGs) were related to the immune response and showed higher expression in the sensitive group. UALCAN showed that high expression of 28 of the top 50 DEGs was associated with a favorable prognosis (p < 0.25), and that this reached a significant (p < 0.05) level for 18 of them, suggesting that patients with high expression of these genes might have benefited from chemotherapy and thus had a better outcome. In preNAC biopsy tissues from a DCF-sensitive case, we demonstrated the presence of cells expressing mRNA for CXCL9, one of the prognosis-related DEGs. Our results highlight the association of immune-related expression profile in preNAC ESCC with the DCF-NAC efficacy.

Supplementary Figure 2 from MicroRNA-375 Is Downregulated in Gastric Carcinomas and Regulates Cell Survival by Targeting PDK1 and 14-3-3ζ

Constitutive activation of the mitogen-activated protein kinase (MAPK) signaling pathway is essential for tumorigenesis of pancreatic ductal adenocarcinoma (PDAC). To date, however, almost all clinical trials of inhibitor targeting this pathway have failed to improve the outcome of patients with PDAC. We found that implanted MIA Paca2, a human PDAC cell line sensitive to a MAPK inhibitor, PD0325901, became refractory within a week after treatment. By comparing the expression profiles of MIA Paca2 before and after acquisition of the refractoriness to PD0325901, we identified clusterin (CLU) as a candidate gene involved. CLU was shown to be induced immediately after treatment with PD0325901 or expressed primarily in more than half of PDAC cell lines, enhancing cell viability by escaping from apoptosis. A combination of PD0325901 and CLU downregulation was found to synergistically or additively reduce the proliferation of PDAC cells. In surgically resected PDAC tissues, overexpression of CLU in cancer cells was observed immunohistochemically in approximately half of the cases studied. Collectively, our findings highlight the mechanisms responsible for the rapid refractory response to MEK inhibitor in PDAC cells, suggesting a novel therapeutic strategy that could be applicable to patients with PDAC using inhibitor targeting the MAPK signaling pathway and CLU.

Supplementary Tables 1-4 from MicroRNA-375 Is Downregulated in Gastric Carcinomas and Regulates Cell Survival by Targeting PDK1 and 14-3-3ζ

Evaluation of Helicobacter pylori resistance to antibiotics is crucial for treatment strategy in Myanmar. Moreover, the genetic mechanisms involved remain unknown. We aimed to investigate the prevalence of H. pylori infection, antibiotic resistance, and genetic mechanisms in Myanmar. One hundred fifty patients from two cities, Mawlamyine (n = 99) and Yangon (n = 51), were recruited. The prevalence of H. pylori infection was 43.3% (65/150). The successfully cultured H. pylori isolates (n = 65) were tested for antibiotic susceptibility to metronidazole, levofloxacin, clarithromycin, amoxicillin, and tetracycline by Etest, and the resistance rates were 80%, 33.8%, 7.7%, 4.6%, and 0%, respectively. In the multidrug resistance pattern, the metronidazole–levofloxacin resistance was highest for double-drug resistance (16/19; 84.2%), and all triple-drug resistance (3/3) was clarithromycin–metronidazole–levofloxacin resistance. Twenty-three strains were subjected to next-generation sequencing to study their genetic mechanisms. Interestingly, none of the strains resistant to clarithromycin had well-known mutations in 23S rRNA (e.g., A2142G, A2142C, and A2143G). New type mutation genotypes such as pbp1-A (e.g., V45I, S/R414R), 23S rRNA (e.g., T248C), gyrA (e.g., D210N, K230Q), gyrB (e.g., A584V, N679H), rdxA (e.g., V175I, S91P), and frxA (e.g., L33M) were also detected. In conclusion, the prevalence of H. pylori infection and its antibiotic resistance to metronidazole was high in Myanmar. The H. pylori eradication regimen with classical triple therapy, including amoxicillin and clarithromycin, can be used as the first-line therapy in Myanmar. In addition, next-generation sequencing is a powerful high-throughput method for identifying mutations within antibiotic resistance genes and monitoring the spread of H. pylori antibiotic-resistant strains.

Background: In the recent studies, a less virulent Helicobacter pylori variant could still colonize the human stomach and induce gastric inflammation, suggesting the involvement of other virulence factors, such as TlyA hemolysin. Nevertheless, the association of TlyA in the pathogenesis of H. pylori infection remains unclear. We investigated the tlyA profile and determined its relationship with gastritis severity. Methods: An observational study was conducted using DNA stocks and secondary data from previous studies. The tlyA variant was examined by NGS and confirmed with polymerase chain reaction. Gastritis severity was categorized by the Updated Sydney System. The relationship between a variant of tlyA and gastritis severity was determined, in which discrete variables were tested using the χ2 test or Fisher exact test. Results: Two H. pylori tlyA variants were observed and characterized as tlyA1 and tlyA2. We noted a unique variant in the amino acid sequence 32–35 that is exclusively detected among H. pylori isolated from the Papua island. In addition, we observed that the tlyA variant had a significant association with the H. pylori density in the antral (p = 0.002). Histological analyses revealed that TlyA1 was associated with higher H. pylori density than TlyA2. However, we did not observe any significant association of tlyA with the infiltration of inflammation cells. Conclusions: We observed 2 tlyA variants (tlyA1 and tlyA2). A significant association of tlyA with bacterial density suggested that tlyA plays a more significant role in the colonization process than its influence on the severity of inflammation in gastric mucosa.

BACKGROUND:Dyspepsia is a frequent main symptom of inpatients and outpatients scenario in Indonesia. However, the number of endoscopy facilities are still low, thus the use of non-invasive method to detect gastritis is necessary. We measured the relationship between urease levels and the stage of gastritis in dyspeptic adult patients.METHODS:A cross-sectional study included outpatient dyspepsia patient from November 2018 to February 2019. We examined 14C-Urea Breath Test (UBT) and determined the stage of gastritis based on the Updated Sydney System classification.RESULTS:The urease level of acute and chronic gastritis positive patients were higher than negative patients (p = 0.001, r = 0.353; p <0.0001, r = 0.433, respectively). The AUC value of 14C-UBT to detect acute, chronic, and atrophic gastritis are 0.889, 0.632 and 0.544, respectively. The best cut-off points of 14C-UBT to predict acute gastritis was ≥26.50δ‰ with sensitivity and specificity being 88.89% and 63.95%, respectively. Whereas the best cut-off points for chronic gastritis was ≥34.50δ‰ with 82.89% sensitivity, 63.16% specificity. As for atrophic gastritis, it showed very low AUC value, hence it is not a sufficient test modality to predict atrophic gastritis cases.CONCLUSION:14C-UBT is sufficient for predicting acute or chronic gastritis but not for atrophic gastritis.

Despite recent advances in sequencing technology and large-scale drug screenings employing hundreds of cell lines, the predictive accuracy of mutation-based biomarkers is still insufficient as a guide for cancer therapy. Therefore, novel types of diagnostic methods using alternative biomarkers would be highly desirable. We have hypothesized that sensitivity-specific changes in the phosphorylation of signaling molecules could be useful in this respect. Here, with the aim of developing a method for predicting the response of cancers to cisplatin using a combination of specific biomarker(s) and patient-derived tumor organoids (PDOs), we found that cisplatin-sensitive cell lines or PDOs showed enhanced phosphorylation of c-Jun (p-c-Jun) within 24 h after cisplatin treatment. We also compared the responses of 6 PDOs to cisplatin with the therapeutic effect of neoadjuvant chemotherapy (docetaxel/cisplatin/5-fluorouracil) in 6 matched patients. Mechanistically, the c-Jun induction was partly related to TNF signaling induced by cisplatin. Our data suggest that enhanced phosphorylation of c-Jun in response to cisplatin treatment could be a predictive biomarker for the efficacy of cisplatin in selected cancer patients. This study shows the potential application of c-Jun phosphorylation upon cisplatin treatment as a biomarker for cisplatin sensitivity in combination with patient-derived tumor organoids (PDOs). We also compared the responses of 6 PDOs to cisplatin with the therapeutic effect of neoadjuvant chemotherapy in 6 matched patients with esophageal squamous cell carcinoma.