Refinement of prognostication for IDH-mutant astrocytomas using DNA methylation-based classification.

The 2021 World Health Organization (WHO) grading system of isocitrate dehydrogenase (IDH)-mutant astrocytomas relies on histological features and the presence of homozygous deletion of the cyclin-dependent kinase inhibitor 2A and 2B (CDKN2A/B). DNA methylation profiling has become highly relevant in the diagnosis of central nervous system (CNS) tumors including gliomas, and it has been incorporated into routine clinical diagnostics in some countries. In this study, we, therefore, examined the value of DNA methylation-based classification for prognostication of patients with IDH-mutant astrocytomas. We analyzed histopathological diagnoses, genome-wide DNA methylation array data, and chromosomal copy number alteration profiles from a cohort of 385 adult-type IDH-mutant astrocytomas, including a local cohort of 127 cases and 258 cases from public repositories. Prognosis based on WHO 2021 CNS criteria (histological grade and CDKN2A/B homozygous deletion status), other relevant chromosomal/gene alterations in IDH-mutant astrocytomas and DNA methylation-based subclassification according to the molecular neuropathology classifier were assessed. We demonstrate that DNA methylation-based classification of IDH-mutant astrocytomas can be used to predict outcome of the patients equally well as WHO 2021 CNS criteria. In addition, methylation-based subclassification enabled the identification of IDH-mutant astrocytoma patients with poor survival among patients with grade 3 tumors and patients with grade 4 tumors with a more favorable outcome. In conclusion, DNA methylation-based subclassification adds prognostic information for IDH-mutant astrocytomas that can further refine the current WHO 2021 grading scheme for these patients.

Isolation, characterization, and immunomodulatory activity evaluation of probiotic strains from colostrum and canine milk.

Background: This study aimed to characterize potential probiotic strains for use in dogs to prevent infectious enteropathies. Lactic acid bacteria (LAB) isolated from canine milk and colostrum were characterized according to their functional properties, including their resistance to gastrointestinal conditions, inhibitory effect against pathogens, and intestinal adhesion. Methods: The immunomodulatory effects of the strains were also analyzed in in vitro and in vivo studies. Among the strains evaluated, two LAB strains (TUCO-16 and TUCO-17) showed remarkable resistance to pH 3.0, bile salts, and pancreatin, as well as inhibitory effects against pathogenic Escherichia coli, Salmonella sp., and Clostridium perfringens. Results: The TUCO-16 and TUCO-17 strains induced a significant increase in the expression of TNF-α, IL-8, and TLR2 in canine macrophages. The oral administration of TUCO-16 and TUCO-17 strains to mice significantly augmented their resistance to pathogenic E. coli or Salmonella intestinal infections. Both canine strains reduced intestinal damage and pathogen counts in the liver and spleen and avoided their dissemination into the bloodstream. These protective effects were related to the ability of TUCO-16 and TUCO-17 strains to differentially modulate the production of IFN-γ, IFN-ß, TNF-α, IL-6, KC, MCP-1, and IL-10 in the intestinal mucosa. Conclusion: Both strains, TUCO-16 and TUCO-17, are potential probiotic candidates for improving intestinal health in dogs, particularly for their ability to inhibit the growth of Gram-negative pathogens common in gastrointestinal infections and modulate the animal's immune response. Further studies are required to effectively demonstrate the beneficial effects of TUCO-16 and TUCO-17 strains in dogs.

Social determinants of the non-use of the explicit health guarantees plan (the GES plan).

INTRODUCTION: The public policy called Explicit health guarantees (GES) could serve as a basis for the future implementation of universal health coverage in Chile. An improvement in the quality of health of the Chilean population has been observed since the launching of the GES, which has a high adherence (84% of the beneficiary population uses this health program). This work seeks the social determinants related to a portion of the remaining 16% of people who do not use the GES. METHODS: This secondary analysis study used a sample of GES recipients (n = 164,786) from the National Socioeconomic Characterization Survey (CASEN) 2020. The GES recipients included in the study responded that they had been under medical treatment for 20 of the 85 pathologies included in the GES, and they had not had access to such policy due to "trust in physician/facility," "decided not to wait," or "lack of information." The CASEN survey chose the 20 pathologies. The Average Marginal Effects of social determinants of the non-use of the GES health plan were predicted using multivariable and panel multinomial probit regression analyses, where the outcome variable assumed three possible values (the three reasons for not accessing) while taking those variables reported in previous studies as independent variables. RESULTS: A higher probability of non-access due to distrust in the physician/facility among adults with higher economic income was found. Among those who prefer not to wait are vulnerable groups of people: women, people with a lower-middle income, those who belong to groups with longer waiting times, and ethnic groups. The people who least access the GES due to lack of information correspond to part of the migrant population and those belonging to the lowest income group. CONCLUSIONS: The GES policy must necessarily improve the timeliness and quality of the services to make them attractive to groups that currently do not have access to them, managing waiting times rather than referrals and using patient-centered evaluations, especially in those most vulnerable groups that do not access GES because they choose not to wait or lack the necessary information, thereby improving their health literacy.

Social determinants of serum 25-hydroxyvitamin D concentrations deficiency in older Chilean people.

Serum 25-hydroxyvitamin D concentrations deficiency is a growing health problem that affects a significant part of the world's population, with particularly negative consequences in children and older adults. Public health has prioritized healthy aging; thus, an investigation of the social determinants related to deficient and insufficient Serum 25-hydroxyvitamin D concentrations in older adults is needed to contribute to the implementation of comprehensive social programs focused on addressing those conditions adversely affecting the health of this group. This study was conducted using a sample of older adults (age ≥ 65 years, n = 1283) from the National Health Survey (NHS 2016-2017). The Average Marginal Effects of the social determinants of Serum 25-hydroxyvitamin D concentrations deficiency in older adults were predicted using a probit model in which the outcome variable assumed two values (deficiency or not deficiency), taking as independent variables those reported in previous studies. The model showed an adequate goodness of fit, Count R2 = 0.65, and the independent variables explained between 11% (Cox-Snell) and 14% (Nagelkerke) of the variance of the outcome variable. The social determinants associated with a greater likelihood of Serum 25-hydroxyvitamin D concentrations deficiency are the following conditions: women, people of native origin, urban dwellers, shorter sunlight exposure, and greater geographical latitude. Implications are discussed, and limitations are considered. Promotion and prevention programs should preferentially target older adults in the southernmost regions who live in urban areas, with a special focus on women. Due to the country's characteristics (17°-57° south latitude), it is necessary to review in future research the three zones shown in this study as relevant social determinants for the older adults living in them to generate inputs in formulating public health policies. The authorities must define the cut-off points for considering the difference between the country's ranges of Serum 25-hydroxyvitamin D concentrations insufficiency and deficiency.

Image-guided radiation therapy (IGRT) in prostate cancer in México, survey of SOMERA (Sociedad Mexicana de Radioterapeutas/Mexican Society of Radiation Oncologists) with recommendations on its implementation and process.

Background: Prostate cancer is one of the main tumors worldwide, its treatment is multidisciplinary, includes radiotherapy in all stages: curative, radical, adjuvant, salvage and palliative. Technological advances in planning systems, image acquisition and treatment equipment have allowed the delivery of higher doses limiting toxicity in healthy tissues, distributing radiation optimally and ensuring reproducibility of conditions. Image-guided radiotherapy (IGRT) is not standard in guidelines, only recommended with heterogeneity in its own process. Materials and methods: A survey was conducted to members of the Mexican Society of Radiation Oncologists (SOMERA), to know the current status and make recommendations about its implementation and use, taking into account existing resources. Results: Responses of 541 patients were evaluated, 85% belonged to the intermediate-high risk group, 65% received adjuvant or salvage radiotherapy (RT), 80% received intensity-modulated radiation therapy (IMRT) using doses up to 80 Gy/2 Gy. Cone beam computed tomography (CBCT) was performed on 506 (93.5%), (100% IMRT) and 90% at a periodicity of 3-5/week. 3D treatment with 42% portal images 1/week. Online correction strategies (36% changes before treatment), following a diet and bladder and rectal control. Evidence and recommendations are reviewed. Conclusions: IGRT should be performed in patients with prostate cancer. In Mexico, despite limitations in the distribution of human and technological resources, it is routinely applied. More information is still needed on clinical evidence of its benefits and the process should be implemented according to infrastructure, following institutional guidelines, recommending to report the initial experience that helps to standardize national conduct.

The clinical value of proneural, classical and mesenchymal protein signatures in WHO 2021 adult-type diffuse lower-grade gliomas.

OBJECTIVES: Accumulating evidence shows that mesenchymal transition of glioblastomas is associated with a more aggressive course of disease and therapy resistance. In WHO2021-defined adult-type diffuse gliomas of lower grade (dLGG), the transition of the tumor phenotype over time, has not been studied. Most efforts to correlate proneural, classical or mesenchymal phenotype with outcome in dLGG were made prior to the WHO 2021 classification. Here, we set out to investigate if phenotype predicted survival and tumor recurrence in a clinical cohort of dLGGs, re-classified according to the 2021 WHO criteria. METHODS: Using a TMA-based approach with five immunohistochemical markers (EGFR, p53, MERTK, CD44 and OLIG2), we investigated 183 primary and 49 recurrent tumors derived from patients with previously diagnosed dLGG. Of the 49 relapses, nine tumors recurred a second time, and one a third time. RESULTS: In total, 71.0% of all tumors could be subtyped. Proneural was most dominant in IDH-mut tumors (78.5%), mesenchymal more common among IDH-wt tumors (63.6%). There was a significant difference in survival between classical, proneural and mesenchymal phenotypes in the total cohort (p<0.001), but not after molecular stratification (IDH-mut: p = 0.220, IDH-wt: p = 0.623). Upon recurrence, proneural was retained in 66.7% of the proneural IDH-mut dLGGs (n = 21), whereas IDH-wt tumors (n = 10) mainly retained or gained mesenchymal phenotype. No significant difference in survival was found between IDH-mut gliomas remaining proneural and those shifting to mesenchymal phenotype (p = 0.347). CONCLUSION: Subtyping into classical, proneural and mesenchymal phenotypes by five immunohistochemical markers, was possible for the majority of tumors, but protein signatures did not correlate with patient survival in our WHO2021-stratified cohort. At recurrence, IDH-mut tumors mainly retained proneural, while IDH-wt tumors mostly retained or gained mesenchymal signatures. This phenotypic shift, associated with increased aggressiveness in glioblastoma, did not affect survival. Group sizes were, however, too small to draw any firm conclusions.

Stalled oligodendrocyte differentiation in IDH-mutant gliomas.

BACKGROUND: Roughly 50% of adult gliomas harbor isocitrate dehydrogenase (IDH) mutations. According to the 2021 WHO classification guideline, these gliomas are diagnosed as astrocytomas, harboring no 1p19q co-deletion, or oligodendrogliomas, harboring 1p19q co-deletion. Recent studies report that IDH-mutant gliomas share a common developmental hierarchy. However, the neural lineages and differentiation stages in IDH-mutant gliomas remain inadequately characterized. METHODS: Using bulk transcriptomes and single-cell transcriptomes, we identified genes enriched in IDH-mutant gliomas with or without 1p19q co-deletion, we also assessed the expression pattern of stage-specific signatures and key regulators of oligodendrocyte lineage differentiation. We compared the expression of oligodendrocyte lineage stage-specific markers between quiescent and proliferating malignant single cells. The gene expression profiles were validated using RNAscope analysis and myelin staining and were further substantiated using data of DNA methylation and single-cell ATAC-seq. As a control, we assessed the expression pattern of astrocyte lineage markers. RESULTS: Genes concordantly enriched in both subtypes of IDH-mutant gliomas are upregulated in oligodendrocyte progenitor cells (OPC). Signatures of early stages of oligodendrocyte lineage and key regulators of OPC specification and maintenance are enriched in all IDH-mutant gliomas. In contrast, signature of myelin-forming oligodendrocytes, myelination regulators, and myelin components are significantly down-regulated or absent in IDH-mutant gliomas. Further, single-cell transcriptomes of IDH-mutant gliomas are similar to OPC and differentiation-committed oligodendrocyte progenitors, but not to myelinating oligodendrocyte. Most IDH-mutant glioma cells are quiescent; quiescent cells and proliferating cells resemble the same differentiation stage of oligodendrocyte lineage. Mirroring the gene expression profiles along the oligodendrocyte lineage, analyses of DNA methylation and single-cell ATAC-seq data demonstrate that genes of myelination regulators and myelin components are hypermethylated and show inaccessible chromatin status, whereas regulators of OPC specification and maintenance are hypomethylated and show open chromatin status. Markers of astrocyte precursors are not enriched in IDH-mutant gliomas. CONCLUSIONS: Our studies show that despite differences in clinical manifestation and genomic alterations, all IDH-mutant gliomas resemble early stages of oligodendrocyte lineage and are stalled in oligodendrocyte differentiation due to blocked myelination program. These findings provide a framework to accommodate biological features and therapy development for IDH-mutant gliomas.

Longitudinal DNA methylation analysis of adult-type IDH-mutant gliomas.

Diffuse gliomas are the most prevalent malignant primary brain tumors in adults and remain incurable despite standard therapy. Tumor recurrence is currently inevitable, which contributes to a persistent high morbidity and mortality in these patients. In this study, we examined the genome-wide DNA methylation profiles of primary and recurrent adult-type IDH-mutant gliomas to elucidate DNA methylation changes associated with tumor progression (with or without malignant transformation). We analyzed DNA methylation profiles of 37 primary IDH-mutant gliomas and 42 paired recurrences using the DNA methylation EPIC beadChip array. DNA methylation-based classification reflected the tumor progression over time. We observed a methylation subtype switch in a proportion of IDH-mutant astrocytomas; the primary tumors were subclassified as low-grade astrocytomas, which progressed to high-grade astrocytomas in the recurrent tumors. The CNS WHO grade 4 IDH-mutant astrocytomas did not always resemble methylation subclasses of higher grades. The number of differentially methylated CpG sites increased over time, and astrocytomas accumulated more differentially methylated CpG sites than oligodendrogliomas during tumor progression. Few differentially methylated CpG sites were shared between patients. We demonstrated that DNA methylation profiles are mostly maintained during IDH-mutant glioma progression, but CpG site-specific methylation alterations can occur.

Social Determinants of the Non-Utilization of the Supplementary Feeding Program (PACAM) Aimed at Older Adults' Nutritional Support.

Chile has implemented the PACAM program to support older people with nutrition and for the prevention of malnutrition and frailty. This work aims to identify the social determinants of older persons not withdrawing PACAM food in order to obtain helpful knowledge for improving the program. First, the CASEN Survey 2017 was used (960,498 observations); the inclusion criterion was PACAM recipients (Yes/No). Next, a probit model was performed with a dichotomous response to determine the marginal effects of each independent variable (e.g., demographic, health, and social). The model shows a good fit (64.4%) with an explained variance between 10.5% to 14.1%. Those variables with more significant marginal effects are people aged 70-75, having tertiary and secondary education, urban living, not participating in social organizations, immigrants, and living in the austral zone. On the other hand, a higher likelihood of consumption was found among people of greater vulnerability (lowest income, lowest education, low health insurance, and aged over 80) and, therefore, in greater fragility. To conclude, the program achieves effective targeting, although improvement actions are required to expand coverage in some groups (indigenous people, immigrants, and people with disabilities). Moreover, authorities should evaluate and reinforce the program with tailored strategies for the older adults who actually withdraw food.

DNA methylation profiling improves routine diagnosis of paediatric central nervous system tumours: A prospective population-based study.

AIMS: Paediatric brain tumours are rare, and establishing a precise diagnosis can be challenging. Analysis of DNA methylation profiles has been shown to be a reliable method to classify central nervous system (CNS) tumours with high accuracy. We aimed to prospectively analyse CNS tumours diagnosed in Sweden, to assess the clinical impact of adding DNA methylation-based classification to standard paediatric brain tumour diagnostics in an unselected cohort. METHODS: All CNS tumours diagnosed in children (0-18 years) during 2017-2020 were eligible for inclusion provided sufficient tumour material was available. Tumours were analysed using genome-wide DNA methylation profiling and classified by the MNP brain tumour classifier. The initial histopathological diagnosis was compared with the DNA methylation-based classification. For incongruent results, a blinded re-evaluation was performed by an experienced neuropathologist. RESULTS: Two hundred forty tumours with a histopathology-based diagnosis were profiled. A high-confidence methylation score of 0.84 or more was reached in 78% of the cases. In 69%, the histopathological diagnosis was confirmed, and for some of these also refined, 6% were incongruent, and the re-evaluation favoured the methylation-based classification. In the remaining 3% of cases, the methylation class was non-contributory. The change in diagnosis would have had a direct impact on the clinical management in 5% of all patients. CONCLUSIONS: Integrating DNA methylation-based tumour classification into routine clinical analysis improves diagnostics and provides molecular information that is important for treatment decisions. The results from methylation profiling should be interpreted in the context of clinical and histopathological information.

Stemness and clinical features in relation to the subventricular zone in diffuse lower-grade glioma: an exploratory study.

Background: The subventricular zone (SVZ) of the human brain is a site of adult stem cell proliferation and a microenvironment for neural stem cells (NSCs). It has been suggested that NSCs in the SVZ are potential cells of origin containing driver mutations of glioblastoma, but their role in the origin of diffuse lower-grade gliomas (dLGGs) is not much studied. Methods: We included 188 patients ≥18 years with IDH-mutated dLGG (WHO grades 2-3) histologically diagnosed between 2007 and 2020. Tissue microarrays of tumor samples for patients between 2007 and 2016 were used for immunodetection of Nestin, SOX2, SOX9, KLF4, NANOG, CD133 cMYC, and Ki67. DNA methylation profile was used for stemness index (mDNAsi). Tumor contact with the SVZ was assessed and the distance was computed. Results: Overall, 70.2% of the dLGG had SVZ contact. Tumors with SVZ contact were larger (102.4 vs 30.9 mL, P < .01), the patients were older (44.3 vs 40.4 years, P = .04) and more often had symptoms related to increased intracranial pressure (31.8% vs 7.1%, P < .01). The expression of SOX2, SOX9, Nestin, and Ki67 showed intersample variability, but no difference was found between tumors with or without SVZ contact, nor with the actual distance to the SVZ. mDNAsi was similar between groups (P = .42). Conclusions: We found no statistical relationship between proximity with the SVZ and mDNAsi or expression of SOX2, SOX9, Nestin, and Ki67 in IDH-mutated dLGG. Our data suggest that the potential impact of SVZ on IDH-mutated dLGG is probably not associated with a more stemness-like tumor profile.

[Social determinants of access to the explicit guarantees health program implemented in Chile]. / Determinantes sociales que influyen en el acceso en Chile al Plan GES, según CASEN 2017.

BACKGROUND: In Chile, an eventual implementation of a plan with universal health coverage is a challenge. The already implemented explicit health guarantees plan (GES) could be a benchmark. For this reason, it is important to obtain information about the results of its implementation. AIM: To identify the social determinants of health that influence the access to GES. MATERIAL AND METHODS: The National Socioeconomic Characterization Survey performed in 2017 was used as a data source. The beneficiaries of 20 diseases covered by GES and inquired in the survey were considered for the present study. RESULTS: People with the higher probability of access to GES plan belong to the lowest income quintiles, are nationals, live in the central-southern metropolitan Santiago, have lower education, have a public health insurance program (FONASA) and are aged mostly over 60 years. The diseases with the highest probability of access to the program are primary arterial hypertension, type 1 and type 2 diabetes mellitus, acute myocardial infarction, moderate and severe bronchial asthma, breast cancer, colon cancer, and bipolar disorder. CONCLUSIONS: The access probability to the GES program is in line with the epidemiological profile of the Chilean population, and with a greater social vulnerability.

DNA methylation alterations across time and space in paediatric brain tumours.

DNA methylation is increasingly used for tumour classification and has expanded upon the > 100 currently known brain tumour entities. A correct diagnosis is the basis for suitable treatment for patients with brain tumours, which is the leading cause of cancer-related death in children. DNA methylation profiling is required for diagnosis of certain tumours, and used clinically for paediatric brain tumours in several countries. We therefore evaluated if the methylation-based classification is robust in different locations of the same tumour, and determined how the methylation pattern changed over time to relapse. We sampled 3-7 spatially separated biopsies per patient, and collected samples from paired primary and relapse brain tumours from children. Altogether, 121 samples from 46 paediatric patients with brain tumours were profiled with EPIC methylation arrays. The methylation-based classification was mainly homogeneous for all included tumour types that were successfully classified, which is promising for clinical diagnostics. There were indications of multiple subclasses within tumours and switches in the relapse setting, but not confirmed as the classification scores were below the threshold. Site-specific methylation alterations did occur within the tumours and varied significantly between tumour types for the temporal samples, and as a trend in spatial samples. More alterations were present in high-grade tumours compared to low-grade, and significantly more alterations with longer relapse times. The alterations in the spatial and temporal samples were significantly depleted in CpG islands, exons and transcription start sites, while enriched in OpenSea and regions not affiliated with a gene, suggesting a random location of the alterations in less conserved regions. In conclusion, more DNA methylation changes accumulated over time and more alterations occurred in high-grade tumours. The alterations mainly occurred in regions without gene affiliation, and did not affect the methylation-based classification, which largely remained homogeneous in paediatric brain tumours.

Spatial heterogeneity in DNA methylation and chromosomal alterations in diffuse gliomas and meningiomas.

Adult-type diffuse gliomas and meningiomas are the most common primary intracranial tumors of the central nervous system. DNA methylation profiling is a novel diagnostic technique increasingly used also in the clinic. Although molecular heterogeneity is well described in these tumors, DNA methylation heterogeneity is less studied. We therefore investigated the intratumor genetic and epigenetic heterogeneity in diffuse gliomas and meningiomas, with focus on potential clinical implications. We further investigated tumor purity as a source for heterogeneity in the tumors. We analyzed genome-wide DNA methylation profiles generated from 126 spatially separated tumor biopsies from 39 diffuse gliomas and meningiomas. Moreover, we evaluated five methods for measurement of tumor purity and investigated intratumor heterogeneity by assessing DNA methylation-based classification, chromosomal copy number alterations and molecular markers. Our results demonstrated homogeneous methylation-based classification of IDH-mutant gliomas and further corroborates subtype heterogeneity in glioblastoma IDH-wildtype and high-grade meningioma patients after excluding samples with low tumor purity. We detected a large number of differentially methylated CpG sites within diffuse gliomas and meningiomas, particularly in tumors of higher grades. The presence of CDKN2A/B homozygous deletion differed in one out of two patients with IDH-mutant astrocytomas, CNS WHO grade 4. We conclude that diffuse gliomas and high-grade meningiomas are characterized by intratumor heterogeneity, which should be considered in clinical diagnostics and in the assessment of methylation-based and molecular markers.

Social determinants of malnutrition in Chilean children aged up to five.

BACKGROUND: This study aimed to ascertain the Social Determinants (SDs) of malnutrition (over and undernutrition) of Chilean children aged up to five. METHODS: The study was carried out using a sample of children from zero to five years old (n = 1,270,485; 52.2% female) from the National Socioeconomic Characterization Survey (CASEN) 2017. A multinomial logistic regression model was used, where the "child nutritional status" outcome variable assumed three possible values: normal nutrition, overnutrition, and undernutrition, while taking those variables reported in previous literature as independent variables. RESULTS: The model, by default, set normal nutrition as the reference group, Count R2 = 0.81. Results show a higher likelihood of both overnutrition and undernutrition among male children from the lowest quintiles, with native ethnic backgrounds, reporting health problems, having public health insurance, and who attend kindergarten. Additionally, higher probabilities of undernutrition in younger than two and living in the north of the country, while overnutrition is more likely in the south. CONCLUSIONS: Socioeconomic variables are fundamentally related to both over and undernutrition; the current single schema program to prevent malnutrition should consider SDs such as ethnicity and geographical location, among others; moreover, successful nutritional programs-which focused on the lowest quintiles, need to be expanded to other vulnerable groups and pay more attention to overnutrition.

Determinantes sociales que influyen en el acceso en Chile al Plan GES, según CASEN 2017 / Social determinants of access to the explicit guarantees health program implemented in Chile

BACKGROUND: In Chile, an eventual implementation of a plan with universal health coverage is a challenge. The already implemented explicit health guarantees plan (GES) could be a benchmark. For this reason, it is important to obtain information about the results of its implementation. AIM: To identify the social determinants of health that influence the access to GES. MATERIAL AND METHODS: The National Socioeconomic Characterization Survey performed in 2017 was used as a data source. The beneficiaries of 20 diseases covered by GES and inquired in the survey were considered for the present study. RESULTS: People with the higher probability of access to GES plan belong to the lowest income quintiles, are nationals, live in the central-southern metropolitan Santiago, have lower education, have a public health insurance program (FONASA) and are aged mostly over 60 years. The diseases with the highest probability of access to the program are primary arterial hypertension, type 1 and type 2 diabetes mellitus, acute myocardial infarction, moderate and severe bronchial asthma, breast cancer, colon cancer, and bipolar disorder. CONCLUSIONS: The access probability to the GES program is in line with the epidemiological profile of the Chilean population, and with a greater social vulnerability.

Lactiplantibacillus plantarum Strains Modulate Intestinal Innate Immune Response and Increase Resistance to Enterotoxigenic Escherichia coli Infection.

Currently, probiotic bacteria with not transferable antibiotic resistance represent a sustainable strategy for the treatment and prevention of enterotoxigenic Escherichia coli (ETEC) in farm animals. Lactiplantibacillus plantarum is among the most versatile species used in the food industry, either as starter cultures or probiotics. In the present work, the immunobiotic potential of L. plantarum CRL681 and CRL1506 was studied to evaluate their capability to improve the resistance to ETEC infection. In vitro studies using porcine intestinal epithelial (PIE) cells and in vivo experiments in mice were undertaken. Expression analysis indicated that both strains were able to trigger IL-6 and IL-8 expression in PIE cells in steady-state conditions. Furthermore, mice orally treated with these strains had significantly improved levels of IFN-γ and TNF-α in the intestine as well as enhanced activity of peritoneal macrophages. The ability of CRL681 and CRL1506 to beneficially modulate intestinal immunity was further evidenced in ETEC-challenge experiments. In vitro, the CRL1506 and CRL681 strains modulated the expression of inflammatory cytokines (IL-6) and chemokines (IL-8, CCL2, CXCL5 and CXCL9) in ETEC-stimulated PIE cells. In vivo experiments demonstrated the ability of both strains to beneficially regulate the immune response against this pathogen. Moreover, the oral treatment of mice with lactic acid bacteria (LAB) strains significantly reduced ETEC counts in jejunum and ileum and prevented the spread of the pathogen to the spleen and liver. Additionally, LAB treated-mice had improved levels of intestinal IL-10 both at steady state and after the challenge with ETEC. The protective effect against ETEC infection was not observed for the non-immunomodulatory TL2677 strain. Furthermore, the study showed that L. plantarum CRL1506 was more efficient than the CRL681 strain to modulate mucosal immunity highlighting the strain specific character of this probiotic activity. Our results suggest that the improved intestinal epithelial defenses and innate immunity induced by L. plantarum CRL1506 and CRL681 would increase the clearance of ETEC and at the same time, protect the host against detrimental inflammation. These constitute valuable features for future probiotic products able to improve the resistance to ETEC infection.

The Respiratory Commensal Bacterium Dolosigranulum pigrum 040417 Improves the Innate Immune Response to Streptococcus pneumoniae.

Previously, we demonstrated that the nasal administration of Dolosigranulum pigrum 040417 differentially modulated the respiratory innate immune response triggered by the activation of Toll-like receptor 2 in infant mice. In this work, we aimed to evaluate the beneficial effects of D. pigrum 040417 in the context of Streptococcus pneumoniae infection and characterize the role of alveolar macrophages (AMs) in the immunomodulatory properties of this respiratory commensal bacterium. The nasal administration of D. pigrum 040417 to infant mice significantly increased their resistance to pneumococcal infection, differentially modulated respiratory cytokines production, and reduced lung injuries. These effects were associated to the ability of the 040417 strain to modulate AMs function. Depletion of AMs significantly reduced the capacity of the 040417 strain to improve both the reduction of pathogen loads and the protection against lung tissue damage. We also demonstrated that the immunomodulatory properties of D. pigrum are strain-specific, as D. pigrum 030918 was not able to modulate respiratory immunity or to increase the resistance of mice to an S. pneumoniae infection. These findings enhanced our knowledge regarding the immunological mechanisms involved in modulation of respiratory immunity induced by beneficial respiratory commensal bacteria and suggested that particular strains could be used as next-generation probiotics.

DNA methylation profiling for molecular classification of adult diffuse lower-grade gliomas.

BACKGROUND: DNA methylation profiling has facilitated and improved the classification of a wide variety of tumors of the central nervous system. In this study, we investigated the potential utility of DNA methylation profiling to achieve molecular diagnosis in adult primary diffuse lower-grade glioma (dLGG) according to WHO 2016 classification system. We also evaluated whether methylation profiling could provide improved molecular characterization and identify prognostic differences beyond the classical histological WHO grade together with IDH mutation status and 1p/19q codeletion status. All patients diagnosed with dLGG in the period 2007-2016 from the Västra Götaland region in Sweden were assessed for inclusion in the study. RESULTS: A total of 166 dLGG cases were subjected for genome-wide DNA methylation analysis. Of these, 126 (76%) were assigned a defined diagnostic methylation class with a class prediction score ≥ 0.84 and subclass score ≥ 0.50. The assigned methylation classes were highly associated with their IDH mutation status and 1p/19q codeletion status. IDH-wildtype gliomas were further divided into subgroups with distinct molecular features. CONCLUSION: The stratification of the patients by methylation profiling was as effective as the integrated WHO 2016 molecular reclassification at predicting the clinical outcome of the patients. Our study shows that DNA methylation profiling is a reliable and robust approach for the classification of dLGG into molecular defined subgroups, providing accurate detection of molecular markers according to WHO 2016 classification.

WHO Grade Loses Its Prognostic Value in Molecularly Defined Diffuse Lower-Grade Gliomas.

BACKGROUND: While molecular insights to diffuse lower-grade glioma (dLGG) have improved the basis for prognostication, most established clinical prognostic factors come from the pre-molecular era. For instance, WHO grade as a predictor for survival in dLGG with isocitrate dehydrogenase (IDH) mutation has recently been questioned. We studied the prognostic role of WHO grade in molecularly defined subgroups and evaluated earlier used prognostic factors in the current molecular setting. MATERIAL AND METHODS: A total of 253 adults with morphological dLGG, consecutively included between 2007 and 2018, were assessed. IDH mutations, codeletion of chromosomal arms 1p/19q, and cyclin-dependent kinase inhibitor 2A/B (CDKN2A/B) deletions were analyzed. RESULTS: There was no survival benefit for patients with WHO grade 2 over grade 3 IDH-mut dLGG after exclusion of tumors with known CDKN2A/B homozygous deletion (n=157) (log-rank p=0.97). This was true also after stratification for oncological postoperative treatment and when astrocytomas and oligodendrogliomas were analyzed separately. In IDH-mut astrocytomas, residual tumor volume after surgery was an independent prognostic factor for survival (HR 1.02; 95% CI 1.01-1.03; p=0.003), but not in oligodendrogliomas (HR 1.02; 95% CI 1.00-1.03; p=0.15). Preoperative tumor size was an independent predictor in both astrocytomas (HR 1.03; 95% CI 1.00-1.05; p=0.02) and oligodendrogliomas (HR 1.05; 95% CI 1.01-1.09; p=0.01). Age was not a significant prognostic factor in multivariable analyses (astrocytomas p=0.64, oligodendrogliomas p=0.08). CONCLUSION: Our findings suggest that WHO grade is not a robust prognostic factor in molecularly well-defined dLGG. Preoperative tumor size remained a prognostic factor in both IDH-mut astrocytomas and oligodendrogliomas in our cohort, whereas residual tumor volume predicted prognosis in IDH-mut astrocytomas only. The age cutoffs for determining high risk in patients with IDH-mut dLGG from the pre-molecular era are not supported by our results.