Low expression of E-Cadherin and CDH1 variants associated with diffuse gastric cancer.

Background: Reduced or null expression of E-cadherin protein is a frequent cause of diffuse gastric cancer (DGC). More than 50% of patients with DGC present somatic variants in CDH1 gene. Objectives: The objectives of this study were to study E-cadherin expression and identify variants in the CDH1 gene in gastric tumors of patients with DGC. Methods: We studied 18 Mexican DGC patients who attended a hospital of the Mexican Social Security Institute; E-cadherin expression was determined by immunohistochemistry, and variants were identified by Sanger sequencing in promoter and coding regions. Predictive analysis was performed using PolyPhen-2 and HOPE software. Results: We found that 56% of DGC patients showed reduced expression of E-cadherin. All patients carried CDH1 variants; overall, 12 different CDH1 variants were identified. Predictive analysis revealed that the rs114265540 variant was probably damaging, with a value of 0.985, indicating a functional impact on the E-cadherin protein. Variants rs34939176 and rs33964119 were identified as risk factors for DGC (odds' ratios [OR] = 31.3, 95% CI 6.3-154.0, p < 0.001; OR = 6.1, 95% CI 2.0-19.0, p < 0.001, respectively) given their elevated frequency and by comparing it with those reported for MXL population in the 1000 Genomes Project database. Conclusions: In this Mexican population, the percentage of diffuse gastric tumors with reduced expression of E-cadherin was similar to that reported in other populations. All gastric tumors of DGC patients studied had somatic CDH1 gene variants; however, the rs114265540, rs34939176, and rs33964119 variants were importantly related to DGC.

New inhibitor targeting Acyl-CoA synthetase 4 reduces breast and prostate tumor growth, therapeutic resistance and steroidogenesis.

Acyl-CoA synthetase 4 (ACSL4) is an isoenzyme of the fatty acid ligase-coenzyme-A family taking part in arachidonic acid metabolism and steroidogenesis. ACSL4 is involved in the development of tumor aggressiveness in breast and prostate tumors through the regulation of various signal transduction pathways. Here, a bioinformatics analysis shows that the ACSL4 gene expression and proteomic signatures obtained using a cell model was also observed in tumor samples from breast and cancer patients. A well-validated ACSL4 inhibitor, however, has not been reported hindering the full exploration of this promising target and its therapeutic application on cancer and steroidogenesis inhibition. In this study, ACSL4 inhibitor PRGL493 was identified using a homology model for ACSL4 and docking based virtual screening. PRGL493 was then chemically characterized through nuclear magnetic resonance and mass spectroscopy. The inhibitory activity was demonstrated through the inhibition of arachidonic acid transformation into arachidonoyl-CoA using the recombinant enzyme and cellular models. The compound blocked cell proliferation and tumor growth in both breast and prostate cellular and animal models and sensitized tumor cells to chemotherapeutic and hormonal treatment. Moreover, PGRL493 inhibited de novo steroid synthesis in testis and adrenal cells, in a mouse model and in prostate tumor cells. This work provides proof of concept for the potential application of PGRL493 in clinical practice. Also, these findings may prove key to therapies aiming at the control of tumor growth and drug resistance in tumors which express ACSL4 and depend on steroid synthesis.

Phenotypical, Clinical, and Molecular Aspects of Adults and Children With Homozygous Familial Hypercholesterolemia in Iberoamerica.

OBJECTIVE: Characterize homozygous familial hypercholesterolemia (HoFH) individuals from Iberoamerica. Approach and Results: In a cross-sectional retrospective evaluation 134 individuals with a HoFH phenotype, 71 adults (age 39.3±15.8 years, 38.0% males), and 63 children (age 8.8±4.0 years, 50.8% males) were studied. Genetic characterization was available in 129 (96%). The majority (91%) were true homozygotes (true HoFH, n=79, 43.0% children, 46.8% males) or compound heterozygotes (compound heterozygous familial hypercholesterolemia, n=39, 51.3% children, 46.2% males) with putative pathogenic variants in the LDLR. True HoFH due to LDLR variants had higher total (P=0.015) and LDL (low-density lipoprotein)-cholesterol (P=0.008) compared with compound heterozygous familial hypercholesterolemia. Children with true HoFH (n=34) tended to be diagnosed earlier (P=0.051) and had a greater frequency of xanthomas (P=0.016) than those with compound heterozygous familial hypercholesterolemia (n=20). Previous major cardiovascular events were present in 25 (48%) of 52 children (missing information in 2 cases), and in 43 (67%) of 64 adults with LDLR variants. Children who are true HoFH had higher frequency of major cardiovascular events (P=0.02), coronary heart (P=0.013), and aortic/supra-aortic valve diseases (P=0.022) than compound heterozygous familial hypercholesterolemia. In adults, no differences were observed in major cardiovascular events according to type of LDLR variant. From 118 subjects with LDLR variants, 76 (64%) had 2 likely pathogenic or pathogenic variants. In 89 subjects with 2 LDLR variants, those with at least one null allele were younger (P=0.003) and had a greater frequency of major cardiovascular events (P=0.038) occurring at an earlier age (P=0.001). CONCLUSIONS: There was a high frequency of cardiovascular disease even in children. Phenotype and cardiovascular complications were heterogeneous and associated with the type of molecular defect.

Predicting Pathogenicity of CDH1 Gene Variants in Patients with Early-onset Diffuse Gastric Cancer from Western Mexico.

BACKGROUND: Early-onset diffuse gastric cancer (EODGC) occurs at or before 50 years of age. Pathogenic mutations and germline deletions in the CDH1 gene (E-cadherin) are well-documented genetic factors associated with the causes of EODGC. OBJECTIVE: The objective of the study was to study CDH1 germline variants and their potential functional impact in patients with EODGC in a Mexican population. METHODS: We studied seven EODGC patients from a biomedical research center in western Mexico. Variants were identified by Sanger sequencing and multiplex ligation-dependent probe amplification. The DeepSEA and SNPClinic v.1.0 software and the Ensembl (1000 Genomes Project, 1kGP) and ClinVar databases were used to predict functional single-nucleotide polymorphisms (SNPs). The genetic admixture of the Mexican patients was corroborated by 22 short tandem repeat loci genotyping and structure analysis. RESULTS: We found 12 germline CDH1 variants in all EODGC patients, and all of them are considered as polymorphisms: rs34561447, rs5030625, rs16260, rs1330727101, rs28372783, rs942269593, rs3743674, rs1801552, rs34939176, rs33964119, rs3556654, and rs1801026. The prediction of regulatory SNPs in the promoter suggests a role for a retrovirus in EODGC that induces the transcription of interferon-related genes through toll-like receptor-interferon response factor 3 signaling, as three SNPs in the CDH1 promoter alter three binding sites for this transcription factor. In addition, SNPs rs28372783 and rs1801026 could alter upstream stimulatory factors 1 (USF1)/USF2-mediated telomerase-dependent lymphocyte activation in EODGC. Other interesting result is a CTCF-dependent shorter CDH1 isoform lacking exon 14, probably due to exon-skipping mediated by rs33964119. CONCLUSIONS: Classical pathogenic germline mutations in the CDH1 gene were not found in these 7 EODGC patients. However, the in silico approaches revealed the possible involvement of a retrovirus and a shorter E-cadherin isoform in EODGC. Nevertheless, further in vitro and in vivo assays are needed to confirm these predictions.

Autosomal dominant early onset Alzheimer's disease in the Mexican state of Jalisco: High frequency of the mutation PSEN1 c.1292C>A and phenotypic profile of patients.

Mutations in three genes (APP, PSEN1, and PSEN2) are the main cause of the autosomal dominant early-onset Alzheimer's disease (AD-EOAD). In PSEN1, the A431E (c.1292C>A, rs63750083) mutation is suspected to have exerted a founder effect in the State of Jalisco, Mexico. In Guadalajara, Jalisco, Mexico, this mutation was found in 46 index cases evaluated for AD-EOAD. In our genealogical analysis, 301 affected relatives of the mutation carriers were identified, 195 of whom were already deceased at the time of interview. Moreover, 560 descendants had a 50% risk of carrying the mutation, and 348 were potentially at risk. A systematic phenotyping was performed in 39 patients. The mean onset age was 42.5 ± 3.9 years, and no significant difference in onset age was observed between the male and female patients. Furthermore, a substantial clinical heterogeneity and high frequencies of spastic paraparesis, language disorders, and neuropsychiatric symptoms were observed. To our knowledge, the investigated families represent the second biggest population carrying a PSEN1 mutation in Latin America, offering a unique opportunity to study the genetic basis of Alzheimer's disease. Addressing AD-EOAD warrants an integral approach involving a deep understanding of its clinical behavior, as well as counseling protocols and prevention studies.

Cost-Effectiveness of the Prenatal Detection of Congenital Heart Diseases: A Systematic Literature Review.

Background: Congenital heart disease is the most common congenital condition worldwide, with a prevalence of 80 cases per 10 000 live births. In addition to perinatal morbidity and mortality, it entails long-term consequences such as multiple surgeries, prolonged hospitalizations, lifelong cardiac follow-up, reduced quality of life, risk of heart failure, and premature mortality in adulthood. This significant health and economic burden on healthcare systems and families highlights the relevance of evaluating the cost-effectiveness of methods for early detection of this condition. Objective: To conduct a systematic literature review (SLR) to identify and analyze existing economic evaluations on prenatal detection of congenital heart diseases through ultrasound, focusing on the reported cost-effectiveness results and the methodological quality of the evaluated studies according to established criteria. Methods: An SLR of economic evaluations was conducted following PRISMA guidelines. A quantitative synthesis of key methodological components of each economic evaluation was performed. The incremental medical costs, effectiveness measures, and cost-effectiveness ratios reported in each study were compiled and compared. The methodological quality was assessed according to compliance with the 24 CHEERS criteria. Results: We found 785 articles, of which only 7 met all inclusion criteria. Most were cost-effectiveness analyses, with the most common outcome being number of cases detected. Screening with only 4-chamber views interpreted by general practitioners or cardiologists were dominant strategies compared with screening with 4-chamber plus outflow views interpreted by a general practitioner. Fetal echocardiography was most effective but most expensive. Screening with 4-chamber and outflow view, followed by referral to a specialist, were recommended as the least expensive strategy per defect detected. On average, articles met 17 of the 24 CHEERS criteria. Discussion: While recent cost-effectiveness analyses demonstrated improved methodological quality, there was a lack of homogeneity due to differences in comparators and population subgroups analyzed. Despite this heterogeneity, fetal ultrasonography screening was consistently identified as a cost-effective strategy, with its cost-effectiveness heavily influenced by the expertise of the interpreting physician. Conclusion: Most studies recommend implementing obstetric ultrasonography screening, without routine fetal echocardiography, for detecting congenital heart diseases.

Clinical and molecular characterization of Costello syndrome in unrelated Mexican patients.

This study intends to describe for the first time a cohort of Mexican patients with Costello syndrome. The five exons of the HRAS gene were amplified in DNA samples from 13 patients with a clinical suspicion of Costello syndrome. PCR products were sequenced using the Ready Reaction Big Dye Terminator v.3.0 Kit and an ABI PRISM 310 sequencer. Only five patients (38%) showed causal variant in codon 12 of the HRAS gene (four with the p.Gly12Ser and one with the p.Gly12Ala variant). Three patients showed silent polymorphic variants (p.His27His and p.Leu159Leu). Clinical features in patients carrying the causal variant were variable. The alternative diagnosis of cardio-facio-cutaneous syndrome was considered in patients who did not have a causative variant in HRAS.

Low Expression of E-Cadherin and Cdh1 Variants Associated with Diffuse Gastric Cancer

ABSTRACT Background Reduced or null expression of E-cadherin protein is a frequent cause of diffuse gastric cancer (DGC). More than 50% of patients with DGC present somatic variants in CDH1 gene. Objectives The objectives of this study were to study E-cadherin expression and identify variants in the CDH1 gene in gastric tumors of patients with DGC. Methods We studied 18 Mexican DGC patients who attended a hospital of the Mexican Social Security Institute; E-cadherin expression was determined by immunohistochemistry, and variants were identified by Sanger sequencing in promoter and coding regions. Predictive analysis was performed using PolyPhen-2 and HOPE software. Results We found that 56% of DGC patients showed reduced expression of E-cadherin. All patients carried CDH1 variants; overall, 12 different CDH1 variants were identified. Predictive analysis revealed that the rs114265540 variant was probably damaging, with a value of 0.985, indicating a functional impact on the E-cadherin protein. Variants rs34939176 and rs33964119 were identified as risk factors for DGC (odds' ratios [OR] = 31.3, 95% CI 6.3-154.0, p < 0.001; OR = 6.1, 95% CI 2.0-19.0, p < 0.001, respectively) given their elevated frequency and by comparing it with those reported for MXL population in the 1000 Genomes Project database. Conclusions In this Mexican population, the percentage of diffuse gastric tumors with reduced expression of E-cadherin was similar to that reported in other populations. All gastric tumors of DGC patients studied had somatic CDH1 gene variants; however, the rs114265540, rs34939176, and rs33964119 variants were importantly related to DGC.

Predicting Pathogenicity of CDH1 Gene Variants in Patients with Early-onset Diffuse Gastric Cancer from Western Mexico

ABSTRACT Background: Early-onset diffuse gastric cancer (EODGC) occurs at or before 50 years of age. Pathogenic mutations and germline deletions in the CDH1 gene (E-cadherin) are well-documented genetic factors associated with the causes of EODGC. Objective: The objective of the study was to study CDH1 germline variants and their potential functional impact in patients with EODGC in a Mexican population. Methods: We studied seven EODGC patients from a biomedical research center in western Mexico. Variants were identified by Sanger sequencing and multiplex ligation-dependent probe amplification. The DeepSEA and SNPClinic v.1.0 software and the Ensembl (1000 Genomes Project, 1kGP) and ClinVar databases were used to predict functional single-nucleotide polymorphisms (SNPs). The genetic admixture of the Mexican patients was corroborated by 22 short tandem repeat loci genotyping and structure analysis. Results: We found 12 germline CDH1 variants in all EODGC patients, and all of them are considered as polymorphisms: rs34561447, rs5030625, rs16260, rs1330727101, rs28372783, rs942269593, rs3743674, rs1801552, rs34939176, rs33964119, rs3556654, and rs1801026. The prediction of regulatory SNPs in the promoter suggests a role for a retrovirus in EODGC that induces the transcription of interferon-related genes through toll-like receptor-interferon response factor 3 signaling, as three SNPs in the CDH1 promoter alter three binding sites for this transcription factor. In addition, SNPs rs28372783 and rs1801026 could alter upstream stimulatory factors 1 (USF1)/USF2-mediated telomerase-dependent lymphocyte activation in EODGC. Other interesting result is a CTCF-dependent shorter CDH1 isoform lacking exon 14, probably due to exon-skipping mediated by rs33964119. Conclusions: Classical pathogenic germline mutations in the CDH1 gene were not found in these 7 EODGC patients. However, the in silico approaches revealed the possible involvement of a retrovirus and a shorter E-cadherin isoform in EODGC. Nevertheless, further in vitro and in vivo assays are needed to confirm these predictions.

Multi-objective optimization of methionine+cysteine levels in diets for growing pigs / Nivel óptimo multiobjetivo de metionina+cisteína en dietas para cerdos en crecimiento / Otimização multiobjetivo do nível de metionina+cisteína em dietas para suínos em crescimento

Abstract Background: The addition of synthetic methionine to growing pig diets should consider the requirements of the methionine+cysteine complex, as most cysteine is obtained through dietary methionine. Objective: To determine an optimal level of methionine+cysteine (Met+Cys) in growing pig diets. Methods: Ninety-nine hybrid (Yorkshire×Landrace×Duroc) barrows were used in a completely randomized design (initial body weight: 25.90±3.99 kg). Nine levels of Met+Cys (0.500, 0.525, 0.550, 0.575, 0.600, 0.625, 0.650, 0.675, and 0.700%) were evaluated during 35 days. To determine a multi-objective optimal level of Met+Cys, growth performance and strongly related carcass traits were evaluated. Results: Average daily feed intake (ADFI) and feed:gain ratio (FGR) showed a linear response to the dietary Met+Cys level (p<0.05), with estimated optimal concentration at 0.700%. Average daily gain (ADG) did not show any effect on the Met+Cys level (p>0.05). Carcass characteristics were not affected by the Met+Cys concentration (p>0.05). The multi-objective optimal level for ADG, ADFI, and FGR was 0.667% Met+Cys. Conclusions: According to our results, 0.667% Met+Cys is the best concentration in diets for growing pigs.

Resumen Antecedentes: Las dietas para cerdos en crecimiento deben adicionarse con metionina sintética para suplir los requerimientos del complejo metionina+cisteína (Met+Cis), pues ellos producen gran parte de la cisteína que requieren a partir de la metionina dietaría. Objetivo: determinar el nivel óptimo multiobjetivo de Met+Cis en dietas para cerdos en crecimiento. Métodos: noventa y nueve machos castrados híbridos (Yorkshire×Landrace×Duroc) se utilizaron en un diseño completamente al azar (peso vivo inicial: 25,90±3,99 kg). Se evaluaron nueve niveles de Met+Cys (0,500, 0,525, 0,550, 0,575, 0,600, 0,625, 0,650, 0,675 y 0,700%) durante 35 d. Para determinar el nivel óptimo multiobjetivo de Met+Cis se consideraron las variables de rendimiento de crecimiento y características de la canal que mantienen una fuerte interrelación. Resultados: el promedio de consumo de alimento diario (ADFI) y la conversión alimenticia (FGR) mostraron una respuesta lineal al nivel de Met+Cis en la dieta (p<0,05), con una concentración óptima estimada en 0,700%. El nivel de Met+Cis no afecto a la ganancia diaria de peso (ADG; p>0,05). La concentración de Met+Cis tampoco afectó las características de la canal (p>0,05). El nivel óptimo multi- objetivo de Met+Cis para ADFI, FGR y ADG fue 0,667%. Conclusión: los resultados indican que 0,667% de Met+Cis es la mejor concentración en la dieta para cerdos en crecimiento.

Resumo Antecedentes: a adição de metionina em dietas para suínos em crescimento deve ser feita para atender às exigências do complexo metionina+cisteína (Met+Cis), porque grande parte da cisteína é obtida através da dieta com metionina. Objetivo: determinar o nível multiobjetivo ótimo de Met+Cis em dietas para suínos em crescimento. Métodos: noventa e nove suínos machos castrados híbridos (Yorkshire×Landrace×Duroc) foram utilizados em delineamento inteiramente casualizado, com peso inicial de 25,90±3,99 kg. Nove níveis de Met+Cis (0,500, 0,525, 0,550, 0,575, 0,600, 0,625, 0,650, 0,675 y 0,700%) foram avaliados em suínos em crescimento por 35 d. Para determinar o nível multiobjetivo ótimo de Met+Cis, foram consideradas as variáveis de comportamento de crescimento e características do canal que mantêm uma forte inter-relação. Resultados: a ingestão média diária de alimentos (ADFI) e a conversão alimentar (FGR) mostraram uma resposta linear ao nível de Met+Cis na dieta (p<0,05), estimando a concentração ótima em 0,700%. O ganho de peso diário (ADG) não mostrou efeito devido ao nível de Met+Cis na dieta (p>0,05). As características da carcaça não foram afetadas pela concentração de Met+Cis na dieta (p>0,05). Com ADFI, FGR e ADG, o nível ótimo de Met+Cis foi obtido, encontrando este nível em 0,667%. Conclusões: os resultados indicam que 0,667% de Met+Cis é a melhor concentração na dieta para suínos em crescimento.

Effectiveness and optimum level of protected methionine in fattening pig diets / Efectividad y nivel óptimo de metionina protegida en dietas para cerdos en engorde / Eficácia e nível ideal de metionina protegida em dietas para suínos de engorda

Abstract Background: Methionine (Met) requirements have not been clearly established for fattening pigs due to their metabolic interrelationships and its bioavailability for protein synthesis. Objective: To determine the optimum level of regular crystalline or protected Met in pig diets from nursery to finishing. Methods: A total of 48 crossbred pigs (11.74±1.72 kg of initial body weight) were used. The treatments consisted of adding four levels (0.00, 0.05, 0.10, 0.15% in addition to dietary content) and two types of Met (regular and protected) to pig diets. Results: Nursery, Finishing I, and II pigs fed protected Met increased daily feed intake (DFI; p≤0.10). Protected Met raised daily weight gain (DWG) in nursery pigs and increased backfat thickness (BT) in nursery and grower pigs (p≤0.10). In Finishing I pigs, protected Met increased DWG and improved carcass characteristics (p≤0.10). In nursery and grower pigs, an extra 0.15% Met decreased feed:gain ratio (FGR; p≤0.10). In grower and Finishing II pigs fed extra 0.05% Met improved DWG and extra 0.10% Met reduced plasma urea concentration (p≤0.10). Conclusions: Feeding protected Met in pig diets increases DWG, DFI and BT. Increasing 0.05-0.15% Met level improves FGR, DWG, potentially reducing nitrogen excretion to the environment.

Resumen Antecedentes: los requerimientos de metionina (Met) para cerdos en crecimiento no han sido claramente establecidos, lo que se debe a sus relaciones metabólicas y su biodisponibilidad para la síntesis proteica. Objetivo: determinar el nivel óptimo de Met regular o protegida en dietas para cerdos en crecimiento. Métodos: Se utilizaron un total de 48 cerdos híbridos (11,74±1,72 kg peso vivo inicial). Los tratamientos consistieron en niveles incrementales (0,00, 0,05, 0,10, 0,15% adicionales al contenido de la dieta) y dos tipos de Met (regular y protegida) en la dieta. Resultados: los cerdos en iniciación, Finalización I, y II, alimentados con Met protegida tuvieron un mayor consumo diario de alimento (DFI; p≤0,10). La Met protegida aumentó la ganancia diaria de peso (DWG) durante la etapa de iniciación, e incrementó el grosor de la grasa dorsal (BT) en iniciación y levante (p≤0,10). Durante Finalización I, la Met protegida aumentó la DWG y mejoró las características de la canal (p≤0,10). Durante iniciación y levante, 0,15% extra de Met disminuyó la conversión alimenticia (FGR; p≤0,10). Los cerdos en levante y Finalización II alimentados con 0,05% extra de Met mejoraron la DWG y con 0.10% extra de Met redujeron la concentración de urea en plasma (p≤0,10). Conclusiones: el uso de Met protegida incrementa DWG, DFI y BT. El aumento del nivel de Met de 0,05-0,15% mejora FGR y DWG, y podria disminuir la excreción de nitrógeno al ambiente.

Resumo Antecedentes: os requisitos de metionina (Met) para suínos de engorda não foram claramente estabelecidos devido às suas relações metabólicas e sua biodisponibilidade para a síntese de proteínas. Objetivo: determinar o nível ideal de Met regular ou protegida em dietas para suínos de engorda. Métodos: foram utilizados 48 suínos híbridos (11,74±1,72 kg de peso vivo inicial). Os tratamentos consistiram em quatro níveis (0,00, 0,05, 0,10, 0,15% mais) e dois tipos (regular e protegida) de Met em dietas para suínos de engorda. Resultados: suínos no início, finalização I e II alimentados com Met protegida aumentaram o consumo diário de ração (DFI; p≤0,10). Met protegido aumentou o ganho de peso diário (DWG) em suínos na iniciação e, aumentou espessura da gordura dorsal (BT) em suínos em iniciação e crescimento (p≤0,10). Nos suínos finalização I, Met protegido aumentou DWG e melhorou as características do canal (p≤0,10). Para suínos de iniciação e crescimento, 0,15% extra de Met diminuiu conversão alimentar (FGR; p≤0,10). No crescimento e finalização II, a adição de 0,05% de Met melhoraram o DWG e com 0,10% reduziram a concentração de uréia (p≤0,10). Conclusões: o uso de Met protegida melhora DWG, DFI e BT. O nível de Met aumentado de 0,05-0,15% melhora FGR e DWG; além disso, a excreção de nitrogênio ambiental pode ser diminuída.