Providing an in vitro depiction of microglial cells challenged with immunostimulatory extracellular vesicles of Naegleria fowleri.

Naegleria fowleri is the causative agent of primary amoebic meningoencephalitis, a rapid and acute infection of the central nervous system with a fatal outcome in >97% of cases. Due to the infrequent report of cases and diagnostic gaps that hinder the possibility of recovering clinic isolates, studies related to pathogenesis of the disease are scarce. However, the secretion of cytolytic molecules has been proposed as a factor involved in the progression of the infection. Several of these molecules could be included in extracellular vesicles (EVs), making them potential virulence factors and even modulators of the immune response in this infection. In this work, we evaluated the immunomodulatory effect of EVs secreted by two clinic isolates of Naegleria fowleri using in vitro models. For this purpose, characterization analyses between EVs produced by both isolates were first performed, for subsequent gene transcription analyses post incubation of these vesicles with primary cultures from mouse cell microglia and BV-2 cells. Analyses of morphological changes induced in primary culture microglia cells by the vesicles were also included, as well as the determination of the presence of nucleic acids of N. fowleri in the EV fractions. Results revealed increased expression of NOS, proinflammatory cytokines IL-6, TNF-α, and IL-23, and the regulatory cytokine IL-10 in primary cultures of microglia, as well as increased expression of NOS and IL-13 in BV-2 cells. Morphologic changes from homeostatic microglia, with small cellular body and long processes to a more amoeboid morphology were also observed after the incubation of these cells with EVs. Regarding the presence of nucleic acids, specific Naegleria fowleri DNA that could be amplified using both conventional and qPCR was confirmed in the EV fractions. Altogether, these results confirm the immunomodulatory effects of EVs of Naegleria fowleri over microglial cells and suggest a potential role of these vesicles as biomarkers of primary acute meningoencephalitis.

The "StemDif Sensor Test": A Straightforward, Non-Invasive Assay to Characterize the Secreted Stemness and/or Differentiation Activities of Tumor-Derived Cancer Cell Lines.

Cancer stem cells are a subpopulation of tumor cells characterized by their ability to self-renew, induce tumors upon engraftment in animals and exhibit strong resistance to chemotherapy and radiotherapy. These cells exhibit numerous characteristics in common with embryonic stem cells, expressing some of their markers, typically absent in non-pathological adult differentiated cells. The aim of this study was to investigate the potential of conditioned media from cancer stem cells to modulate the fate of Leukemia Inhibitory Factor (LIF)-dependent murine embryonic stem cells (mESCs) as a way to obtain a direct readout of the secretome of cancer cells. A functional assay, "the StemDif sensor test", was developed with two types of cancer stem cells derived from grade IV glioblastoma (adult and pediatric) or from gastric adenocarcinoma. We show that conditioned media from the selection of adult but not pediatric Glioma-Inducing Cells (GICs) maintain mESCs' pluripotency in correlation with LIF secretion and activation of STAT3 protein. In contrast, conditioned media from gastric adenocarcinoma cells display LIF-independent stemness and differentiation activities on mESC. Our test stands out for its user-friendly procedures, affordability and straightforward output, positioning it as a pioneering tool for in-depth exploration of cancer stem cell secretome characteristics.

Helicobacter pylori infection induces gastric precancerous lesions and persistent expression of Angpt2, Vegf-A and Tnf-A in a mouse model.

Introduction: Helicobacter pylori colonizes the gastric mucosa and induces chronic inflammation. Methods: Using a mouse model of H. pylori-induced gastritis, we evaluated the mRNA and protein expression levels of proinflammatory and proangiogenic factors, as well as the histopathological changes in gastric mucosa in response to infection. Five- to six-week-old female C57BL/6N mice were challenged with H. pylori SS1 strain. Animals were euthanized after 5-, 10-, 20-, 30-, 40- and 50-weeks post infection. mRNA and protein expression of Angpt1, Angpt2, VegfA, Tnf-α, bacterial colonization, inflammatory response and gastric lesions were evaluated. Results: A robust bacterial colonization was observed in 30 to 50 weeks-infected mice, which was accompanied by immune cell infiltration in the gastric mucosa. Compared to non-infected animals, H. pylori-colonized animals showed an upregulation in the expression of Tnf-A, Angpt2 and VegfA at the mRNA and protein levels. In contrast, Angpt1 mRNA and protein expression was downregulated in H. pylori-colonized mice. Conclusion: Our data show that H. pylori infection induces the expression of Angpt2, Tnf-A and Vegf-A in murine gastric epithelium. This may contribute to the pathogenesis of H. pylori-associated gastritis, however the significance of this should be further addressed.

Stable production of recombinant SARS-CoV-2 receptor-binding domain in mammalian cells with co-expression of a fluorescent reporter and its validation as antigenic target for COVID-19 serology testing.

SARS-CoV-2 receptor binding domain (RBD) recognizes the angiotensin converting enzyme 2 (ACE2) receptor in host cells that enables infection. Due to its antigenic specificity, RBD production is important for development of serological assays. Here we have established a system for stable RBD production in HEK 293T mammalian cells that simultaneously express the recombinant fluorescent protein dTomato, which enables kinetic monitoring of RBD expression by fluorescence microscopy. In addition, we have validated the use of this recombinant RBD in an ELISA assay for the detection of anti-RBD antibodies in serum samples of COVID-19 convalescent patients. Recombinant RBD generated using this approach can be useful for generation of antibody-based therapeutics against SARS-CoV-2, as well serological assays aimed to test antibody responses to this relevant virus.

Cultivo primario de Helicobacter pylori a partir de biopsias gástricas obtenidas por endoscopia / Primary culture of Helicobacter pylori from endoscopic gastric biopsies

Objetivo: Determinar la viabilidad del cultivo de la bacteria Helicobacter pylori en Costa Rica por medio de la documentación de toma de muestras, la comparación del diagnóstico histopatológico y la descripción de los diagnósticos asociados a los aislamientos obtenidos con los resultados de la ureasa rápida. Métodos: Investigación descriptiva que involucró a pacientes de entre los 35 y 70 años, de ambos sexos, que asistieron al Servicio de Endoscopia Digestiva del Hospital Clínica Bíblica entre febrero y junio del 2019 para estudio gastroscópico. Se obtuvieron biopsias gástricas para diagnóstico histopatológico, prueba de ureasa rápida y cultivo de Helicobacter pylori. Para este último, se transportaron las biopsias en un medio de transporte semisólido, se maceró el tejido y se cultivó enagar Skirrow y agar selectivo para Helicobacter; una placa de cada medio se incubó a 37 °C en microaerofilia entre 48 horas y 10 días. La positividad del cultivo se realizó por observación de la morfología colonial y la bacteria se identificó por análisis microscópico al fresco, tinción de Gram y pruebas bioquímicas (catalasa, ureasa y oxidasa). Resultados: Se incluyó a 44 pacientes (edad: 50.6 ± 10.0, 54.5% masculinos). Se recuperó Helicobacter pylori en biopsias de 27 pacientes (61.4% de éxito). La recuperación de la bacteria fue similar en el medio Skirrow y en el selectivo para Helicobacter. El porcentaje de éxito de recuperación semanal aumentó durante el estudio hasta alcanzar un éxito del 100% en la semana 11. Se comparó el cultivo con la ureasa rápida en 27 pacientes y la concordancia entre ambos métodos fue de un coeficiente kappa de Cohen de 0.48. El cultivo detectó la bacteria en un 56% de los pacientes, la ureasa rápida en un 37% y la combinación de ambas técnicas permitió la detección en un 60%. El diagnóstico endoscópico más frecuente en los pacientes con cultivo positivo fue la gastritis eritematosa y gastritis crónica superficial y el diagnóstico histopatológico predominante fue gastritis crónica con atrofia gástrica. El diagnóstico por cultivo coincidió con la detección por azul de toluidina en un 80.4% de los casos. Conclusiones: Se puede implementar el cultivo de Helicobacter pylori en Costa Rica. Este estudio tuvo un porcentaje de recuperación de la bacteria de 61.4%. La combinación del método de cultivo con la prueba de ureasa rápida y la detección histológica contribuye a un diagnóstico certero y oportuno. Recomendamos que, con base en protocolos descritos en esta investigación, cada laboratorio estandarice las condiciones que le permitan un buen porcentaje de recuperación y una implementación adaptada a sus actividades de rutina.

Aim: To document the recent experiences on the implementation of sampling and culturing Helicobacter pylori in Costa Rica, to compare it with other diagnostic methods: rapid urease test and histopathology and to describe the diagnoses associated with the obtained isolates. Methods: Descriptive research involving patients who visited the digestive endoscopy department of the Clínica Bíblica hospital in San José, Costa Rica between February and July of 2019 for gastroscopy. Gastric biopsies were obtained and histopathological analysis, rapid urease test, and bacteriological culture for Helicobacter pylori were performed. For culture techniques, the sample was transported in an in-house semi-solid medium. Biopsy fragments were macerated and plated on Skirrow agar and Helicobacterselective in-house agar, and incubated in microaerophilic atmosphere for 48 hours to 10 days. Culture positivity was determined by observation of the colonial morphology and microscopic observation; Gram staining and biochemical tests (urease, catalase, and oxidase) were used for bacterial identification. Results: 44 patients (mean aged 50.6 ± 10.0 years old, 54.5% male) were included in the study. Helicobacter pylori was recovered in biopsies from 27 patients (61.4% success rate). Bacterial growth was similar regardless the culture medium, but the physiological state of the bacteria was better in the Helicobacter-selective agar than in Skirrow. The weekly recovery rate increased to reach a 100% recovery plateau on week 11. Culture was compared with the rapid urease test in 27 patients, and the concordance between both methods using Cohen's kappa coefficient was 0.48. Whilst the culture detected Helicobacter pylori in 56% of the patients, and the rapid urease test in 37%, the combination of both allowed a 60% rate. The most frequent endoscopic diagnosis in patients with positive cultures were erythematous gastritis and chronic superficial gastritis, and the predominant histopathological diagnosis was chronic atrophic gastritis. Culture-based diagnosis was consistent with the histopathology detection of Helicobacter pylori in 80.4% of the cases. Conclusions: The implementation of H. pylori culture in Costa Rica is possible. This study had a 61.4% recovery rate. The combination of culture with rapid urease test and histopathology increases the probability of an accurate and timely diagnosis. We recommend that, based on previously described protocols such as ours, each laboratory adjusts the conditions to allow a good recovery rate and implement H. pylori diagnostic methods most suitable to their routine activities.

Exploratory multigroup structural equation modeling approach to analyzing H. pylori infection, serological pepsinogen levels, and life style factors in an elderly population in Costa Rica / Enfoque exploratorio del modelo de ecuaciones estructurales multigrupo para el análisis de la infección con H. pylori, niveles de pepsinógenos séricos y factores asociados a estilo de vida en una población de adultos mayores en Costa Rica

Abstract The aim of this work is to describe and analyze the association of PGI/PGII ratio (indicator of gastric atrophy) with H. pylori-CagA and life style factors such as caloric intake, obesity, and harmful habits amongst H. pylori-positive elderly people infected in Costa Rica using an exploratory multigroup structural equations model (SEM). Using a sample of 1748 H. pylori-positive elderly people from CRELES first wave study, a SEM was employed analyze if the relationships between PGI/PGII ratio with levels of H. pylori-CagA, caloric intake, obesity, and harmful habits, differs by sex, age and risk areas subgroups. The proposed SEMs exhibited a good fit in males (RMSEA = 0.039), females (RMSEA = 0.000), low-risk area (RMSEA = 0.038), middle-risk area (RMSEA = 0.042), individuals under 80 years (RMSEA = 0.038) and individuals aged 80 and over (RMSEA = 0.042), while an acceptable fit was observed for the high-risk area (RMSEA = 0.061). Fitted SEMs showed that CagA predicted PG-ratio as expected, with effects increasing with the risk area, but similar between sex and age groups. All indicators measuring obesity (BMI, arms, and waist) showed significant standardized coefficients, with similar effects between sex, age and risk area groups. No other significant effects or differences between groups were identified. We propose a good-fitted SEM model for the possible relationships between CagA and PG ratio and the geographical risk area level for elderly people. No differences were observed on measured parameters between male and female population, or between under 80 years and older individuals.

Resumen El objetivo de este trabajo es describir y analizar la asociación entre PGI/PGII (indicador de atrofia gástrica con H. pylori-CagA y factores asociados a estilo de vida como ingesta calórica, obesidad y hábitos nocivos entre adultos mayores positivos por H. pylori en Costa Rica utilizando modelos de ecuaciones estructurales multigrupo (SEM). Con una muestra de 1748 adultos mayores del estudio CRELES, se utilizó un SEM para analizar las relaciones entre PGI/PGII, CagA, ingesta calórica, obesidad y hábitos nocivos difieren por sexo, edad y áreas de riesgo. Los SEMs propuestos exhibieron un buen ajuste en hombres (RMSEA = 0.039), mujeres (RMSEA = 0.000), área de bajo riesgo (RMSEA = 0.038), áreas de riesgo medio (RMSEA = 0.042), individuos menores de 80 años (RMSEA = 0.038) e individuos de 80 años o más (RMSEA = 0.042), mientras que hubo un ajuste aceptable en áreas de alto riesgo (RMSEA = 0.061). Los SEMs ajustados mostraron que CagA predice la relación PGI/II en la dirección esperada con efectos proporcionales al área de riesgo, pero no por sexo y edad. Todos los indicadores medibles de obesidad (IMC, brazos y cintura) mostraron coeficientes estandarizados significativos con efectos similares entre los grupos por sexo, edad y área de riesgo. No se encontraron otros efectos o diferencias significativas. Proponemos un modelo SEM bien ajustado para las posibles relaciones entre CagA y PGI/II y el nivel de riesgo del área geográfica en adultos mayores. No se encontraron diferencias en las variables analizadas entre hombres y mujeres ni entre los grupos de edad.

The TNF-A-857*T Polymorphism is Associated with Gastric Adenocarcinoma Risk in a Costa Rican Population.

BACKGROUND: Costa Rica is ranked as one of the countries with highest incidence of gastric cancer worldwide. Previous studies in Costa Rican populations have revealed associations between gastric cancer risk and several cytokine polymorphisms that seem to play a role in the regulation of the expression of these proteins. In this study, we assessed associations of the polymorphisms IL-6-174 G/C, IFNGR1-56 C/T, IL-8-251 T/A and TNF-A (-857 C/T, -308 A/G) with gastric pathologies in a high-risk population of Latin America. METHODS: DNA samples of 47 patients with gastric adenocarcinoma, 53 with chronic gastritis, 56 with duodenal ulcer and 94 healthy controls, were genotyped for the five mentioned SNPs. All participants were ≥50-years-old. Genotyping was performed by PCR-RFLP and 5'-nuclease PCR assay. H. pylori infection, CagA status, pepsinogen (PG) I and II blood levels were determined by ELISA. Logistic regression analysis was used to determine possible associations of the polymorphisms with cancer, gastritis and duodenal ulcer, and linear regression analysis to determine associations with blood PG levels. RESULTS: A total of 86.6% of the population was positive for H. pylori; of them, 51.6% was CagA+. Patients with the TNF-A-857*T allele had an increased risk for gastritis (OR: 3.67, p = 0.015) and gastric adenocarcinoma (OR:6.15, p = 0.001). Associations between other polymorphisms and gastric diseases, or PG levels, were not found. CONCLUSIONS: Our results indicate that the TNF-A-857*T SNP is among the risk factors associated with the risk of gastric cancer in Costa Rica.

Primary Amebic Meningoencephalitis Related to Groundwater in Costa Rica: Diagnostic Confirmation of Three Cases and Environmental Investigation.

During the first trimester of 2020, the Ministry of Health of Costa Rica reported the first three cases of primary amebic meningoencephalitis (PAM). In two cases, laboratory personnel of the hospitals preliminarily identified amoeboid forms in cerebrospinal fluid (CSF) samples. For the molecular confirmation of species, CSF samples were sent to our laboratory. We carried out microscopic analyses and exflagellation assays. Besides, samples were cultured in 2% casein hydrolysate medium and in non-nutrient agar plates supplemented with Escherichia coli. Finally, PCR and sequencing were employed for the molecular diagnosis and species identification. In all cases, the presence of Naegleria fowleri was confirmed. An environmental investigation to identify the possible infection sources was also performed. Water samples from hot springs and groundwater from an artisan well were collected and after filtration and culture in non-nutrient agar plates supplemented with E. coli, thermotolerance and exflagellation assays were carried out. For the positive samples, PCR and sequencing were performed, confirming the presence of N. fowleri in several water samples. The report of these cases and the possible association with hot springs has had a significant impact on the population and health authorities of Costa Rica.

Leukaemia Inhibitory Factor (LIF) Inhibits Cancer Stem Cells Tumorigenic Properties through Hippo Kinases Activation in Gastric Cancer.

Cancer stem cells (CSCs) present chemo-resistance mechanisms contributing to tumour maintenance and recurrence, making their targeting of utmost importance in gastric cancer (GC) therapy. The Hippo pathway has been implicated in gastric CSC properties and was shown to be regulated by leukaemia inhibitory factor receptor (LIFR) and its ligand LIF in breast cancer. This study aimed to determine LIF's effect on CSC properties in GC cell lines and patient-derived xenograft (PDX) cells, which remains unexplored. LIF's treatment effect on CSC markers expression and tumoursphere formation was evaluated. The Hippo kinase inhibitor XMU-MP-1 and/or the JAK1 inhibitor Ruxolitinib were used to determine Hippo and canonical JAK/STAT pathway involvement in gastric CSCs' response to LIF. Results indicate that LIF decreased tumorigenic and chemo-resistant CSCs, in both GC cell lines and PDX cells. In addition, LIF increased activation of LATS1/2 Hippo kinases, thereby decreasing downstream YAP/TAZ nuclear accumulation and TEAD transcriptional activity. LIF's anti-CSC effect was reversed by XMU-MP-1 but not by Ruxolitinib treatment, highlighting the opposite effects of these two pathways downstream LIFR. In conclusion, LIF displays anti-CSC properties in GC, through Hippo kinases activation, and could in fine constitute a new CSCs-targeting strategy to help decrease relapse cases and bad prognosis in GC.

TAZ Controls Helicobacter pylori-Induced Epithelial-Mesenchymal Transition and Cancer Stem Cell-Like Invasive and Tumorigenic Properties.

Helicobacter pylori infection, the main risk factor for gastric cancer (GC), leads to an epithelial-mesenchymal transition (EMT) of gastric epithelium contributing to gastric cancer stem cell (CSC) emergence. The Hippo pathway effectors yes-associated protein (YAP) and transcriptional co-activator with PDZ binding motif (TAZ) control cancer initiation and progression in many cancers including GC. Here, we investigated the role of TAZ in the early steps of H. pylori-mediated gastric carcinogenesis. TAZ implication in EMT, invasion, and CSC-related tumorigenic properties were evaluated in three gastric epithelial cell lines infected by H. pylori. We showed that H. pylori infection increased TAZ nuclear expression and transcriptional enhancer TEA domain (TEAD) transcription factors transcriptional activity. Nuclear TAZ and zinc finger E-box-binding homeobox 1 (ZEB1) were co-overexpressed in cells harboring a mesenchymal phenotype in vitro, and in areas of regenerative hyperplasia in gastric mucosa of H. pylori-infected patients and experimentally infected mice, as well as at the invasive front of gastric carcinoma. TAZ silencing reduced ZEB1 expression and EMT phenotype, and strongly inhibited invasion and tumorsphere formation induced by H. pylori. In conclusion, TAZ activation in response to H. pylori infection contributes to H. pylori-induced EMT, invasion, and CSC-like tumorigenic properties. TAZ overexpression in H. pylori-induced pre-neoplastic lesions and in GC could therefore constitute a biomarker of early transformation in gastric carcinogenesis.

Verteporfin targeting YAP1/TAZ-TEAD transcriptional activity inhibits the tumorigenic properties of gastric cancer stem cells.

Gastric carcinomas (GC) are heterogeneous tumors, composed of a subpopulation of cluster of differentiation-44 (CD44)+ tumorigenic and chemoresistant cancer stem cells (CSC). YAP1 and TAZ oncoproteins (Y/T) interact with TEA domain family member 1 (TEAD) transcription factors to promote cell survival and proliferation in multiple tissues. Their activity and role in GC remain unclear. This work aimed to analyze Y/T-TEAD activity and molecular signature in gastric CSC, and to assess the effect of verteporfin, a Food and Drug Administration-approved drug preventing Y/T-TEAD interaction, on gastric CSC tumorigenic properties. Y/T-TEAD molecular signature was investigated using bioinformatical (KmPlot database), transcriptomic and immunostaining analyses in patient-derived GC and cell lines. Verteporfin effects on Y/T-TEAD transcriptional activity, CSC proliferation and tumorigenic properties were evaluated using in vitro tumorsphere assays and mouse models of patient-derived GC xenografts. High expressions of YAP1, TAZ, TEAD1, TEAD4 and their target genes were associated with low overall survival in nonmetastatic human GC patients (n = 444). This Y/T-TEAD molecular signature was enriched in CD44+ patient-derived GC cells and in cells resistant to conventional chemotherapy. Verteporfin treatment inhibited Y/T-TEAD transcriptional activity, cell proliferation and CD44 expression, and decreased the pool of tumorsphere-forming CD44+ /aldehyde dehydrogenase (ALDH)high gastric CSC. Finally, verteporfin treatment inhibited GC tumor growth in vivo; the residual tumor cells exhibited reduced expressions of CD44 and ALDH1, and more importantly, they were unable to initiate new tumorspheres in vitro. All these data demonstrate that Y/T-TEAD activity controls gastric CSC tumorigenic properties. The repositioning of verteporfin targeting YAP1/TAZ-TEAD activity could be a promising CSC-based strategy for the treatment of GC.

The Hippo Kinase LATS2 Controls Helicobacter pylori-Induced Epithelial-Mesenchymal Transition and Intestinal Metaplasia in Gastric Mucosa.

BACKGROUND & AIMS: Gastric carcinoma is related mostly to CagA+-Helicobacter pylori infection, which disrupts the gastric mucosa turnover and elicits an epithelial-mesenchymal transition (EMT) and preneoplastic transdifferentiation. The tumor suppressor Hippo pathway controls stem cell homeostasis; its core, constituted by the large tumor suppressor 2 (LATS2) kinase and its substrate Yes-associated protein 1 (YAP1), was investigated in this context. METHODS: Hippo, EMT, and intestinal metaplasia marker expression were investigated by transcriptomic and immunostaining analyses in human gastric AGS and MKN74 and nongastric immortalized RPE1 and HMLE epithelial cell lines challenged by H pylori, and on gastric tissues of infected patients and mice. LATS2 and YAP1 were silenced using small interfering RNAs. A transcriptional enhanced associated domain (TEAD) reporter assay was used. Cell proliferation and invasion were evaluated. RESULTS: LATS2 and YAP1 appear co-overexpressed in the infected mucosa, especially in gastritis and intestinal metaplasia. H pylori via CagA stimulates LATS2 and YAP1 in a coordinated biphasic pattern, characterized by an early transient YAP1 nuclear accumulation and stimulated YAP1/TEAD transcription, followed by nuclear LATS2 up-regulation leading to YAP1 phosphorylation and targeting for degradation. LATS2 and YAP1 reciprocally positively regulate each other's expression. Loss-of-function experiments showed that LATS2 restricts H pylori-induced EMT marker expression, invasion, and intestinal metaplasia, supporting a role of LATS2 in maintaining the epithelial phenotype of gastric cells and constraining H pylori-induced preneoplastic changes. CONCLUSIONS: H pylori infection engages a number of signaling cascades that alienate mucosa homeostasis, including the Hippo LATS2/YAP1/TEAD pathway. In the host-pathogen conflict, which generates an inflammatory environment and perturbations of the epithelial turnover and differentiation, Hippo signaling appears as a protective pathway, limiting the loss of gastric epithelial cell identity that precedes gastric carcinoma development.

Virulence factor genotyping of Helicobacter pylori isolated from Costa Rican dyspeptic patients.

BACKGROUND AND AIMS: Costa Rica is one of the countries with the highest incidence and mortality rates for gastric cancer. Helicobacter pylori infection rates are high in the whole country. We have previously shown that H. pylori CagA+ is significantly associated with atrophic gastritis (AG) of the antrum in a dyspeptic population. The aim of this work is to determine if other H. pylori virulence factors (vacA, dupA, oipA, iceA and babA2) are associated with atrophic gastritis (AG) or duodenal ulcer (DU). METHODS: The presence of virulence genes in Costa Rican H. pylori isolates was analyzed by PCR in 151 cultured strains from patients with dyspeptic symptoms. Endoscopic and histopathological diagnoses were available. Odds-ratio and 95% confidence intervals for AG patients vs. non-atrophic gastritis (NAG) or DU patients vs. no duodenal ulcer (NDU) patients were calculated. RESULTS: Amongst the studied isolates, 82% had the cagA+, 76.2% had the vacA s1m1, 97.0% had the oipA+, 21.0% had the icea1, 79.0% had the iceA2, 44.0% had the babA2+ and 76.0% the dupA+ genotypes. Infection with H pylori cagA+, dupA+, oipA+, iceA, babA2+, and vacA s1m1 genotypes was not associated with AG risk. The frequency of the dupA gene was 78.7 and 60.9% in isolates from patients with NDU and DU, respectively, and its presence was significantly associated with decreased risk of duodenal ulcer [odds-ratio: 0.33, p = 0.024, confidence interval 95% (0.11-0.85)]. CONCLUSION: H. pylori dupA genotype is inversely associated with DU risk in this population.

Priming the seed: Helicobacter pylori alters epithelial cell invasiveness in early gastric carcinogenesis.

Helicobacter pylori (H. pylori) infection is a well-established risk factor for the development of gastric cancer (GC), one of the most common and deadliest neoplasms worldwide. H. pylori infection induces chronic inflammation in the gastric mucosa that, in the absence of treatment, may progress through a series of steps to GC. GC is only one of several clinical outcomes associated with this bacterial infection, which may be at least partially attributed to the high genetic variability of H. pylori. The biological mechanisms underlying how and under what circumstances H. pylori alters normal physiological processes remain enigmatic. A key aspect of carcinogenesis is the acquisition of traits that equip preneoplastic cells with the ability to invade. Accumulating evidence implicates H. pylori in the manipulation of cellular and molecular programs that are crucial for conferring cells with invasive capabilities. We present here an overview of the main findings about the involvement of H. pylori in the acquisition of cell invasive behavior, specifically focusing on the epithelial-to-mesenchymal transition, changes in cell polarity, and deregulation of molecules that control extracellular matrix remodeling.

Gastric cancer: Basic aspects.

Gastric cancer is one of the most incident and deadliest malignancies in the world. Gastric cancer is a heterogeneous disease and the end point of a long and multistep process, which results from the stepwise accumulation of numerous (epi)genetic alterations, leading to dysregulation of oncogenic and tumor suppressor pathways. Gastric cancer stem cells have emerged as fundamental players in cancer development and as contributors to gastric cancer heterogeneity. For this special issue, we will report last year's update on the gastric cancer molecular classification, and in particular address the gastric cancer groups who could benefit from immune checkpoint therapy. We will also review the latest advances on gastric cancer stem cells, their properties as gastric cancer markers and therapeutic targets, and associated signaling pathways. The understanding of the molecular basis underlying gastric cancer heterogeneity and of the role played by gastric cancer stem cells in cancer development and heterogeneity is of major significance, not only for identifying novel targets for cancer prevention and treatment, but also for clinical management and patient stratification for targeted therapies.

The geographic origin of Helicobacter pylori isolated from Costa Rican patients.

Helicobacter pylori infects a significant proportion of the world population and it is associated with pathologies which include chronic atrophic gastritis, peptic ulcer, and gastric neoplasias such as gastric adenocarcinoma and MALT lymphoma. Costa Rica has a high prevalence of the infection and an elevated incidence of gastric cancer and its associated mortality. The global population structure for H. pylori has been established using a MLST scheme. The population structure of the strains of H. pylori circulating in Costa Rica is currently unknown. We characterized the geographical origin of 24 H. pylori isolates from Costa Rican patients. We identified 142 new alleles for the genes included in the scheme and in eight of the 24 isolates from Costa Rican patients, all seven alleles sequenced were described for the first time. Twenty-one isolates from Costa Rican patients group with hpEurope strains and the remaining three isolates grouped with hspWAfrica isolates (Bayesian posterior probability values above 0.70, P = 0.05, after 2 000 000 generations). The obtained result in the MLST analysis was not unexpected and reflects the genetic composition of the Costa Rican population.