Gene expression during the development of Mycobacterium smegmatis biofilms on hydroxyapatite surfaces / Expresión génica durante el desarrollo de biopelículas de Mycobacterium smegmatis en superficies de hidroxiapatita

Bacterial biofilms are a consortium of bacteria that are strongly bound to each other and the surface on which they developed irreversibly. Bacteria can survive adverse environmental conditions and undergo changes when transitioning from a planktonic form to community cells. The process of mycobacteria adhesion is complex, involving characteristics and properties of bacteria, surfaces, and environmental factors; therefore, the formation of different biofilms is possible. Cell wall-, lipid-, and lipid transporter-related genes (glycopeptidolipids, GroEL1, protein kinase) are important in mycobacterial biofilm development. We investigated gene expression during in vitro development of Mycobacterium smegmatis biofilms on a hydroxyapatite (HAP) surface. Biofilm formation by M. smegmatis cells was induced for 1, 2, 3, and 5 days on the HAP surface. Mycobacteria on polystyrene generated an air–liquid interface biofilm, and on the fifth day, it increased by 35% in the presence of HAP. Six genes with key roles in biofilm formation were analyzed by real-time RT‒qPCR during the biofilm formation of M. smegmatis on both abiotic surfaces. The expression of groEL1, lsr2, mmpL11, mps, pknF, and rpoZ genes during biofilm formation on the HAP surface did not exhibit significant changes compared to the polystyrene surface. These genes involved in biofilm formation are not affected by HAP.(AU)

Endoplasmic reticulum stress-related genes construct a risk score and prognostic model for squamous lung cancer in female players

Objective: To investigate the prognostic impact of endoplasmic reticulum stress-related genes on lung squamous carcinoma. Methods: Gene expression data were obtained from the TCGA database, and prognostic models were constructed by LASSO-COX analysis. Gene expression data were obtained from the TCGA database and prognostic models were constructed by LASSO-COX analysis. Immunohistochemistry was performed to validate the gene expression. Subtypes were obtained by clustering analysis and correlation analysis of subtypes was performed. The accuracy of the model was reflected by Monogram. Result: Endoplasmic reticulum stress-related genes LRRK2 and WFS1 constructed the prognostic model. the ROC curve was 0.584 at -1 year, 0.598 at 2 years and 0.603 at 3 years. The risk score constructed from the prognostic model was an independent predictor of overall survival and influenced the tumor microenvironment as well as drug sensitivity. Conclusions: Endoplasmic reticulum stress-related genes can be used to predict the prognosis and influence the immune status of squamous lung cancer, and modulating the expression of these genes is a potential therapeutic option. (AU)

Fear stress promotes glioma progression through inhibition of ferroptosis by enhancing FSP1 stability

Purpose Patients diagnosed with cancer often suffer from emotional stressors, such as anxiety, depression, and fear of death. However, whether fear stress could influence the glioma progression is still unclear. Methods Xenograft glioma animal models were established in nude mice. Tumor-bearing mice were subjected to fear stress by living closely with cats and then their depressive behaviors were measured using an open field test. Hematoxylin and eosin staining, the TUNEL staining and immunochemical staining were used to detect the histopathological changes of tumor tissues. Gene expression profiling was used to screen the aberrant gene expression. Methylated RNA immunoprecipitation was used to identify the RNA m6A level. Gene expression was measured by western blot and real-time PCR, respectively. Results We found that fear stress promoted glioma tumor progression in mice. Fear stress-induced upregulation of METTL3 and FSP1, increased m6A level of glioma tumor tissues, and inhibited ferroptosis in glioma progression, which were reversed by knockdown of METTL3 and FSP1 in vivo. In addition, we found that when iFSP1 (a ferroptosis inducer by targeting inhibition of FSP1) was introduced to glioma cells, the cells viability of glioma significantly was decreased and ferroptosis was enhanced in glioma cells. Conclusions Fear stress-induced upregulation of METTL3 stabilized FSP1 mRNA by m6A modification, leading to tumor progression through inhibition of ferroptosis. Our study provides a new understanding of psychological effects on glioma development, and new insights for glioma therapy (AU)

A review of literature: role of long noncoding RNA TPT1-AS1 in human diseases

Human diseases are multifactorial processes mainly driven by the intricate interactions of genetic and environmental factors. Long noncoding RNAs (lncRNAs) represent a type of non-coding RNAs with more than 200 nucleotides. Multiple studies have demonstrated that the dysregulation of lncRNAs is associated with complex biological as well as pathological processes through various mechanism, especially the regulation of gene transcription and related signal transduction pathways. Moreover, an increasing number of studies have explored lncRNA-based clinical applications in different diseases. For instance, the lncRNA Tumor Protein Translationally Controlled 1 (TPT1) Antisense RNA 1 (TPT1-AS1) was found to be dysregulated in several types of disease and strongly associated with patient prognosis and diverse clinical features. Recent studies have also documented that TPT1-AS1 modulates numerous biological processes through multiple mechanisms, including cell proliferation, apoptosis, autophagy, invasion, migration, radiosensitivity, chemosensitivity, stemness, and extracellular matrix (ECM) synthesis. Furthermore, TPT1-AS1 was regarded as a promising biomarker for the diagnosis, prognosis and treatment of several human diseases. In this review, we summarize the role of TPT1-AS1 in human diseases with the aspects of its expression, relevant clinical characteristics, molecular mechanisms, biological functions, and subsequent clinical applications (AU)

LncRNA NCK1-AS1-mediated regulatory functions in human diseases

Disease development requires the activation of complex multi-factor processes involving numerous long noncoding RNAs (lncRNAs), which describe non-protein-coding RNAs longer than 200 nucleotides. Emerging evidence indicates that lncRNAs act as essential regulators that perform pivotal roles in the pathogenesis and progression of human diseases. The mechanisms underlying lncRNA involvement in diverse diseases have been extensively explored, and lncRNAs are considered powerful biomarkers for clinical practice. The lncRNA noncatalytic region of tyrosine kinase adaptor protein 1 (NCK1) antisense 1 (NCK1-AS1), also known as NCK1 divergent transcript (NCK1-DT), is encoded on human chromosome 3q22.3 and produces a 27,274-base-long transcript. NCK1-AS1 has increasingly been characterized as a causative agent for multiple diseases. The abnormal expression and involvement of NCK1-AS1 in various biological processes have been associated with several diseases. Further exploration of the mechanisms through which NCK1-AS1 contributes to disease development and progression will provide a foundation for potential clinical applications of NCK1-AS1 in the diagnosis and treatment of various diseases. This review summarizes the current understanding of the various functions and mechanisms through which NCK1-AS1 contributes to various diseases and the clinical application prospects for NCK1-AS1 (AU)

Identification of biomarkers and candidate small-molecule drugs in lipopolysaccharide (LPS)-induced acute lung injury by bioinformatics analysis

Background/objective: Acute lung injury (ALI) is a critical clinical syndrome with high rates of incidence and mortality. However, its molecular mechanism remains unclear. The current work aimed to explore the molecular mechanisms of ALI by identifying different expression genes (DEGs) and candidate drugs using a combination of chip analysis and experimental validation. Methods: Three microarray datasets were downloaded from Gene Expression Omnibus (GEO) database to obtain DEGs. We conducted a Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway-enrichment analyses of overlapping DEGs among three databases. The expression level of key gene was verified by Western blotting analysis in LPS-treated ALI cell models. Finally, we predicted the candidate drugs targeting the key gene that might be effective for ALI treatment, and the role of candidate drug in treating ALI was verified by investigation. Results: A total 29 overlapping DEGs were up-regulated in LPS-induced ALI groups. They were enriched in inflammation and inflammation-related pathways. Serpin family A member 3 (SERPINA3) was defined as a key gene because it was associated with inflammation pathway and up-regulated in microarray datasets in LPS-induced ALI. In LPS-induced human bronchial epithelial cells transformed with Ad12-SV40-2B (BEAS-2B) cells, SERPINA3 was enhanced. Pyridoxal phosphate as an upstream drug of SERPINA3 could improve cell viability and reduce expression inflammatory factors in LPS-treated BEAS-2B cells. Conclusion: Our study suggested that pyridoxal phosphate could be a candidate drug targeting SERPINA3 gene in LPS-induced ALI. It has protective and anti-inflammatory effects in BEAS-2B cells, and may become a potential novel treatment for ALI (AU)

Alteraciones de la expresión génica en la infección por SARS-CoV-2 en distintos modelos de estudio / Alterations of gene expression in SARS-CoV-2 infection in different study models

Objetivo: La pandemia por SARS-CoV-2 ha supuesto un auténtico reto para el mundo científico debido a la rápida transmisión y elevada mortalidad que produce este nuevo coronavirus. La enfermedad asociada se ha denominado COVID-19 y abarca desde casos asintomáticos hasta graves que evolucionan rápidamente a síndrome de distrés respiratorio agudo, alteraciones multisistémicas y la muerte. La comunidad científica ha aunado esfuerzos para tratar de conocer mejor el proceso fisiopatológico de la infección con la intención de combatir de forma más eficaz la enfermedad. En este trabajo presentamos un estudio para conocer las alteraciones de la expresión génica provocadas por la infección. Metodología: Se han usado tres modelos de estudio distintos: cultivos de células epiteliales bronquiales, organoides de las vías respiratorias y muestras obtenidas de autopsias en pacientes, con y sin infección por SARS-CoV-2. Se han analizado los perfiles de expresión alterados por la infección en cada modelo, así como las categorías funcionales enriquecidas. Resultados: Solo 4 genes son comunes en los tres tipos de modelos de estudio, siendo el modelo de autopsias el más dispar. Dentro de los genes comunes en los modelos de cultivo celular y organoide de pulmón encontramos funciones relacionadas con procesos inflamatorios. Conclusiones: Los estudios in vitro son un buen modelo para tener una foto fija de las alteraciones en los patrones de infección, mientras que las autopsias no son un buen modelo debido al sesgo provocado por la necrosis. (AU)

Short summary: Three different study models have been used to study the gene expression profiles produced by SARS-CoV-2: bronchial epithelial cell cultures, airway organoids, and autopsy samples from patients with and without SARS-infection. CoV-2. Basis: The SARS-CoV-2 pandemic has been a real challenge for the scientific world due to the rapid transmission and high mortality caused by this new coronavirus. The associated disease has been named COVID-19 and ranges from asymptomatic to severe cases that rapidly progress to acute respiratory distress syndrome, multisystem disorders, and death. The scientific community has joined efforts to try to better understand the pathophysiological process of the infection with the intention of combating the disease more effectively. In this work we present a study to determine the alterations in gene expression caused by the infection. Methods: Three different study models have been used: bronchial epithelial cell cultures, airway organoids, and samples obtained from autopsies in patients with and without SARS-CoV-2 infection. The expression profiles altered by the infection in each model have been analyzed, as well as the functional categories enriched. Results: Only 4 genes are common in the three types of study models, the autopsy model being the most disparate. Within the common genes in cell and organoid culture models of the lung, we find functions related to inflammatory processes. Conclusions: In vitro studies are a good model to have a snapshot of alterations in infection patterns, while autopsies are not a good model due to bias caused by necrosis. (AU)

Relation of beclin-1 and bcl-2 expressions with pathological parameters and prognosis in clear cell renal cell carcinomas

Introduction: The most common type of renal cancers is the clear cell renal cell carcinoma (CCRCC) and 98% of CCRCCshave a loss of sequence in the short arm of chromosome 3 by deletion or translocation. Programmed cell death; another possiblemechanism of tumorigenesis, comprises two separate components: apoptosis and autophagy. This study aims to show the relation between the prognostic parameters and survival, and Beclin-1, as the representative marker of autophagy, and Bcl-2 as therepresentative marker of apoptosis in CCRCC patients. In this study, we aimed to determine if Beclin-1 and Bcl-2 expressionlevels can provide any prognostic information about CCRCC patients.Methods: We examined a total of 84 patients who underwent partial or radical nephrectomy and were diagnosed as havingCCRCC between January 2008 and December 2015. Immunohistochemical staining was performed, the evaluation was forBeclin-1 and Bcl-2 semi-quantitative, and based on the percentage of positively stained cells (proportion) and staining intensity.Results: There was only a statistical significance between Beclin-1 expression and age (r:-0.274; p=0.012; p <0.05). There wasa marginal significance between ISUP grade and Beclin-1 (p=0.051). The relation of Bcl-2 expression with the ISUP grade,recurrence, metastasis, and mortality revealed statistical significance (p=0.001, p=0.019, p=0.009, p=0.013, respectively). TheISUP grade and the Bcl-2 expression revealed statistical significance on multivariate analysis ( HR 7.453, 95% CI: 1.935-28.713,p=0.004). The 5-year and 10-year tumor recurrences rates were lower in Bcl-2 positive group, and Bcl-2 positive group experienced longer disease free and overall survival...(AU)

Dietary interventions in mice affect oxidative stress and gene expression of the Prlr and Esr1 in the adipose tissue and hypothalamus of dams and their offspring

Maternal diet is key to the progeny’s health since it may impact on the offspring’s adult life. In this study, mice dams received standard (CONT), restrictive (RD), or hypercaloric (HD) diets during mating, pregnancy, and lactation. Male offspring of each group of dams also received these diets: CONT, RD, HD. Aiming to evaluate the oxidative stress in the adipose tissue, reactive oxygen species (ROS) production, catalase (CAT), and superoxide dismutase (SOD) activities were analyzed in dams and offspring. In the adipose tissue and hypothalamus, gene expression of prolactin (Prlr) and estrogen alpha (Esr1) receptors was performed in dams and offspring. Protein expression of Stat5 was evaluated in the adipose tissue of the offspring from RD-fed dams. HD-fed dams increased triglycerides and leptin serum concentrations, and decreased SOD activity in the adipose tissue. In the offspring’s adipose tissue, we observed a maternal diet effect caused by HD, with increased ROS production and SOD and CAT activities. Gene expression of Prlr and Esr1 in the offspring’s adipose tissue was decreased due to maternal RD. Mice from HD-fed dams showed higher Stat5 expression compared to the offspring from CONT and RD dams in the adipose tissue. In the hypothalamus, we found decreased expression of Prlr in RD and HD dams, compared to CONT; and a maternal diet effect on Prlr and Esr1 gene expression in the offspring. In conclusion, we can affirm that maternal nutrition impacts the redox state and influences the gene expression of Prlr and Esr1, which are involved in energy metabolism, both peripherally and centrally in the adult life of the female offspring. (AU)

Relación de la expresión génica de IP-10 con la actividad del lupus eritematoso sistémico / Relationship of IP-10 gene expression to systemic lupus erythematosus activity

Objetivo: Evaluar la expresión génica del gen IP-10 en pacientes con lupus eritematoso sistémico (LES) y su posible relación con la actividad de la enfermedad. Pacientes y métodos: El estudio incluyó 120 pacientes diagnosticados con LES y 30 controles sanos. Se investigó la expresión génica relativa de IP-10 con el método fold change, la cual fue correlacionada con el nivel de actividad lúpica evaluado con el instrumento SLEDAI 2-K. Resultados: Se encontraron diferentes niveles en la expresión génica de IP-10 relacionada con la actividad lúpica (p =<0,001). Estos fueron mayores en los pacientes con actividad grave respecto a aquellos sin actividad, baja y moderada. El incremento en la expresión génica del grupo con actividad grave fue significativo con un fold change de tres. Conclusión: El incremento significativo en la expresión génica relativa IP-10 puede ser un marcador de actividad lúpica grave.(AU)

Objectives: To evaluate IP-10 gene expression in patients with SLE, and its possible relationship with disease activity. Patients and methods: This study included 120 patients diagnosed with SLE and 30 healthy controls. The relative gene expression of IP-10 was investigated with the Fold Change method, which was correlated with the level of lupus activity evaluated with the SLEDAI 2-K instrument. Results: Different levels of gene expression were found according to the SLE activity (p =<0.001). IP-10 gene expression levels were higher in patients with severe activity than in those with no activity, low activity, and moderate activity. The increase in gene expression in the severe activity group was significant with a Fold Change of 3. Conclusion: The significant increase in relative gene expression IP-10 may be a marker of severe lupus activity.(AU)

Variabilidad de la expresión clínica de la encefalopatía KCNB1 / Variability of the clinical expression of KCNB1 encephalopathy

Introducción: El gen KCNB1 codifica un canal de potasio dependiente del voltaje que regula corrientes transmembrana en las neuronas piramidales. Variantes en heterocigosis se han asociado recientemente con encefalopatías epilépticas de inicio precoz y discapacidad intelectual, pero su caracterización clínica no está completamente definida.Objetivo: Describir el espectro clínico asociado con variantes de KCNB1 en pacientes pediátricos. Pacientes y métodos: Estudio retrospectivo de cuatro pacientes procedentes de tres familias con encefalopatía KCNB1, analizando características clínicas y electroencefalográficas de la epilepsia, manifestaciones neurológicas asociadas y patrón de neurodesarrollo. Resultados: En dos, la mutación en KCNB1 fue de novo; las otras dos, hermanas, heredaron la variante de un progenitor con mosaicismo germinal. Todos presentaban discapacidad intelectual leve-moderada; dos pacientes, trastorno del espectro autista; y otros dos, trastorno por déficit de atención/hiperactividad. Sólo el caso 2 mostro´ alteraciones en la resonancia magnética cerebral: atrofia cortical evolutiva. Tres desarrollaron epilepsia (casos 1-3). Caso 1: inicio a los 9,5 meses con síndrome de West bien controlado con vigabatrina y zonisamida. Caso 2: inicio a los 13 meses con síndrome de West; desarrollo evolutivo de crisis polimorfas (atónicas, hipermotoras, disautonómicas y tónicas) refractarias a 10 fármacos antiepilépticos y corticoides. Asocio´ trastorno del movimiento caracterizado por ataxia, discinesias y temblor. Caso 3: inicio a los 14,5 años con crisis atónicas, patrón multifocal en el electroencefalograma y adecuado control con levetiracetam. Conclusiones: La encefalopatía KCNB1 presenta una evolución natural heterogénea, principalmente respecto a la epilepsia, y se observan desde pacientes con epilepsia refractaria hasta pacientes sin crisis epilépticas...(AU)

Introduction: The KCNB1 gene encodes a voltage-dependent potassium channel that regulates transmembrane currents in pyramidal neurons. Heterozygous variants have recently been associated with early-onset epileptic encephalopathies and intellectual disability, but their clinical characterisation has not yet been fully defined. Aim: To describe the clinical spectrum associated with variants of KCNB1 in paediatric patients. Patients and methods. Retrospective study of four patients from three families with KCNB1 encephalopathy, including an analysis of the clinical and electroencephalographic features of epilepsy, associated neurological manifestations and neurodevelopmental pattern. Results: In two of them, the mutation in KCNB1 was de novo; the other two, who were sisters, inherited the variant from a parent with germline mosaicism. All had mild-to-moderate intellectual disability, two patients had autistic spectrum disorder and two had attention deficit hyperactivity disorder. Only case 2 displayed alterations in the MRI brain scan: progressive cortical atrophy. Three of them developed epilepsy (cases 1-3). Case 1: onset at 9.5 months with West syndrome that was well controlled with vigabatrine and zonisamide. Case 2: onset at 13 months with West syndrome, evolutionary development of polymorphic seizures (atonic, hypermotor, dysautonomic and tonic) that were refractory to 10 antiepileptic drugs and corticosteroids. Accompanied by a movement disorder characterised by ataxia, dyskinesias and tremor. Case 3: onset at 14.5 years with atonic seizures, multifocal EEG pattern and adequate control with levetiracetam.Conclusions: KCNB1 encephalopathy has a heterogeneous natural history, mainly with respect to epilepsy, ranging from patients with refractory epilepsy to patients without any epileptic seizures. All had neurodevelopmental disorders, such as intellectual disability or autism spectrum disorder, independent of epilepsy.(AU)

Physio-pathological effects of m6A modification and its potential contribution to melanoma

Methylation of N6-adenosine (m6A) is the most prevalent internal RNA modification and is especially common among the messenger RNAs. These m6A modifications regulate splicing, translocation, stability and translation of RNA through dynamic and reversible interactions with m6A-binding proteins, namely the writers, erasers and readers. RNA methyltransferases catalyze the m6A modifications, while demethylases reverse this methylation. Deregulation of the m6A modification process has been implicated in human carcinogenesis, including melanoma—which carries one of the highest mutant rates. In this review, we provide an up-to-date summary of m6A regulation and its biological impacts on normal and cancer cells, with emphasis on the deregulation of m6A modification and m6A regulators in melanoma. In addition, we highlight the prospective potential of exploiting m6A modification in the treatment of melanoma and non-cancer diseases. (AU)

PD-1 and PD-L1 gene expressions and their association with Epstein-Barr virus infection in chronic lymphocytic leukemia

PurposeThe PD-1 (programmed cell death-1) receptor is expressed on the surface of activated T cells. Its ligand, programmed cell death ligand-1 (PD-L1), is expressed on the surface of dendritic cells or macrophages. The PD-1/PD-L1 interaction ensures prevention of autoimmunity by activating the immune system only when needed. In cancers, PD-L1 expressed on the tumour cells binds to PD-1 receptors on the activated T cells, leading to inhibition of the cytotoxic T cells and immunosuppression. PD-1/PD-L1 pathway is upregulated in EBV infection that is known to worsen the CLL prognosis. Therefore, we aimed to study the association between PD-1 and PD-L1 expressions, EBV status and the CLL prognosis.Methods and patientsThe study was conducted on 80 newly diagnosed CLL patients and 80 controls. We analyzed PD-1 and PD-L1 expressions and EBV-DNA load by real-time PCR. The cytogenetic abnormalities and expression of ZAP70 and CD38 were detected by FISH and Flow cytometry, respectively.ResultsPD-1/PD-L1 expressions were significantly upregulated in CLL patients compared to controls. In addition, their mRNA levels were significantly higher in EBV( +) versus EBV( −) patients. High expression of PD-1/PD-L1 was associated with poor prognostic markers (RAI stages of CLL, del 17p13, ZAP70, and CD38 expression), failure of complete remission, shorter progression-free survival, and overall survival.ConclusionHigh expression of PD-1 and PD-L1, together with high EBD-DNA load were linked to worse prognosis in CLL. In addition, PD-1 and PD-L1 might represent suitable therapeutic targets for patients suffering from aggressive CLL. (AU)

Tumor microenvironment-associated gene C3 can predict the prognosis of colorectal adenocarcinoma: a study based on TCGA

Background Colorectal cancer is one of the most common malignancies. With continuous exploration of the interaction between tumor cells and the immune system, tumor immunotherapy has become a revolution. However, CRC remains one of the less effective tumors for immunotherapy. The tumor microenvironment plays an important role in tumorigenesis and progression. The aim of this study is to explore tumor microenvironment-related genes that can predict the prognosis of colorectal adenocarcinoma, and also to provide new ideas for the mechanism of tumor development as well as immunotherapy. Methods After estimating Stromalscore and Immunescore of colorectal adenocarcinoma tumor samples according to RNA-Seq expression data downloaded from TCGA, we screened for TME-related differential genes. We filtered prognosis-related core genes by constructing protein–protein interaction networks and making one-factor cox analysis for prognosis. Finally, the relative content of 22 immune cells in tumor tissues was evaluated, and then immune cells associated with core genes were identified. Results We screened 773 differential genes related to the TME. Then we identified C3 as a core gene associated with prognosis. Single gene analysis showed that C3 expression was significantly higher in tumor tissues than in normal tissues (p < 0.001). High C3 expression was associated with lower overall survival (p = 0.046). Tumor immune cell analysis showed that mast cells resting, mast cells activated, T cells CD4 memory activated, eosinophils, and macrophages M0 were C3-associated immune cells. Conclusions C3 has potential as a biomarker for colorectal adenocarcinoma and could provide new research ideas for the diagnosis and treatment of colorectal adenocarcinoma, especially for immunotherapy (AU)

Biopsia con aguja gruesa para la determinación del perfil de expresión génica tumoral, mediante la plataforma de 70 genes / Core needle biopsy for determining tumour gene expression profile through the 70 gene signature

Objetivos Evaluar la utilidad de la biopsia ecoguiada con aguja gruesa diagnóstica (BAG1) para la determinación del perfil de expresión génica tumoral (PEG), en los tumores malignos de mama. Materiales y métodos Revisión de 130 biopsias ecoguiadas con aguja gruesa (BAG), con resultado de malignidad. Se consideraron «aptas» las muestras con, al menos, un 30% de células tumorales. Se estudió la influencia del tamaño tumoral (menor de 1 cm, entre 1-2 cm y mayor de 2 cm) y se analizaron las causas que motivaron muestras no aptas. Se utilizó la plataforma MammaPrint® (70 genes). Se evaluó la influencia del grado histológico y del riesgo genómico en los resultados. Resultados En la BAG1 se obtuvieron muestras aptas en 100 biopsias (76,92%). Entre los 36 casos en los que se utilizó la BAG para obtener el PEG, en 32 (88,89%) se realizó a partir de la BAG1. Entre los 30 casos en los que la BAG1 no resultó apta, en 26 casos no se obtuvo el porcentaje mínimo de células tumorales en la muestra. Ni el grado histológico ni el riesgo genómico influyeron en los resultados. Conclusiones Las muestras diagnósticas de la biopsia ecoguiada con aguja gruesa (BAG1) pueden ser válidas para la determinación del perfil de expresión génica. Ello facilita y acelera el proceso de evaluación pronóstica en los tumores infiltrantes de mama. Por ello, proponemos que, de manera rutinaria y ante el diagnóstico de tumor maligno infiltrante, conste el porcentaje de células tumorales en los informes anatomopatológicos. (AU)

Objectives To assess the utility of diagnostic ultrasound-guided core needle biopsy (CNB1) for determining tumour gene expression profile (GEP) in malignant breast tumours. Materials and Methods Review of 130 diagnostic ultrasound-guided core needle biopsies (CNB1) indicating malignancy. Samples with at least 30% tumour cells were considered suitable. The influence of tumour size (less than 1 cm, between 1-2 cm and greater than 2 cm) was studied and the causes of unsuitable samples were analysed. The MammaPrint® platform (70 genes) was used. The influence of histological grade and genomic risk was evaluated. Results Suitable CNB1 samples were obtained in 100 biopsies (76.92%). Among the 36 cases in which CNB was used to obtain the GEP, in 32 (88.89%) it was performed using the CNB1. Among the 30 cases in which CNB1 was not suitable, the minimum percentage of tumour cells in the sample was not obtained in 26 cases. Neither histological grade nor genomic risk influenced the results. Conclusions Diagnostic samples from ultrasound-guided biopsy (CNB1) can be valid to determine GEP. This facilitates and accelerates the prognostic evaluation process in infiltrating breast tumours. Therefore, we propose that, when diagnosing an infiltrating malignant tumour, the percentage of tumour cells should be routinely recorded in the pathology reports. (AU)

Evaluation of miR-210 expression in common variable immunodeficiency: patients with unsolved genetic defect

Background Common variable immunodeficiency (CVID) is one of the most prevalent forms of primary immunodeficiency diseases (PID). CVID is characterized by failure in the final differentiation of B lymphocytes and impaired antibody production but the pathogenesis is not known in the majority of patients. We postulated that the expression pattern of miRNAs in unsolved CVID patients might be the underlying epigenetic cause of the disease. Therefore, we aimed to assess the expression of hsa-miR-210-5p and FOXP3 transcription factor in CVID cases in comparison with healthy individuals. Methods Eleven CVID cases with no genetic defects (all PID known genes excluded) and 10 sex and age-matched healthy individuals were enrolled in the study. T lymphocytes were purified from PBMC, and expression levels of miR-210-5p and FOXP3 mRNA were evaluated by real-time PCR. Results We demonstrated that miR-210 expression in patients was significantly higher than the control group (P = 0.03). FOXP3 expression was slightly lower in patients compared with healthy controls (P = 0.86). There was a negative correlation between miR and gene expression (r: −0.11, P = 0.73). Among various clinical complications, autoimmunity showed a considerable rate in high-miR patients (P = 0.12, 42.8%), while autoimmunity was not observed in normal miR-210 patients. Conclusions Our results suggest a role for miR-210 in the pathogenesis of autoimmunity in CVID patients. Further studies would better elucidate epigenetic roles in CVID patients with no genetic defects (AU)

Biopsia con aguja gruesa para la determinación del perfil de expresión génica tumoral, mediante la plataforma de 70 genes / Core needle biopsy for determining tumour gene expression profile through the 70 gene signature

Objetivos Evaluar la utilidad de la biopsia ecoguiada con aguja gruesa diagnóstica (BAG1) para la determinación del perfil de expresión génica tumoral (PEG), en los tumores malignos de mama. Materiales y métodos Revisión de 130 biopsias ecoguiadas con aguja gruesa (BAG), con resultado de malignidad. Se consideraron «aptas» las muestras con, al menos, un 30% de células tumorales. Se estudió la influencia del tamaño tumoral (menor de 1 cm, entre 1-2 cm y mayor de 2 cm) y se analizaron las causas que motivaron muestras no aptas. Se utilizó la plataforma MammaPrint® (70 genes). Se evaluó la influencia del grado histológico y del riesgo genómico en los resultados. Resultados En la BAG1 se obtuvieron muestras aptas en 100 biopsias (76,92%). Entre los 36 casos en los que se utilizó la BAG para obtener el PEG, en 32 (88,89%) se realizó a partir de la BAG1. Entre los 30 casos en los que la BAG1 no resultó apta, en 26 casos no se obtuvo el porcentaje mínimo de células tumorales en la muestra. Ni el grado histológico ni el riesgo genómico influyeron en los resultados. Conclusiones Las muestras diagnósticas de la biopsia ecoguiada con aguja gruesa (BAG1) pueden ser válidas para la determinación del perfil de expresión génica. Ello facilita y acelera el proceso de evaluación pronóstica en los tumores infiltrantes de mama. Por ello, proponemos que, de manera rutinaria y ante el diagnóstico de tumor maligno infiltrante, conste el porcentaje de células tumorales en los informes anatomopatológicos. (AU)

Objectives To assess the utility of diagnostic ultrasound-guided core needle biopsy (CNB1) for determining tumour gene expression profile (GEP) in malignant breast tumours. Materials and Methods Review of 130 diagnostic ultrasound-guided core needle biopsies (CNB1) indicating malignancy. Samples with at least 30% tumour cells were considered suitable. The influence of tumour size (less than 1 cm, between 1-2 cm and greater than 2 cm) was studied and the causes of unsuitable samples were analysed. The MammaPrint® platform (70 genes) was used. The influence of histological grade and genomic risk was evaluated. Results Suitable CNB1 samples were obtained in 100 biopsies (76.92%). Among the 36 cases in which CNB was used to obtain the GEP, in 32 (88.89%) it was performed using the CNB1. Among the 30 cases in which CNB1 was not suitable, the minimum percentage of tumour cells in the sample was not obtained in 26 cases. Neither histological grade nor genomic risk influenced the results. Conclusions Diagnostic samples from ultrasound-guided biopsy (CNB1) can be valid to determine GEP. This facilitates and accelerates the prognostic evaluation process in infiltrating breast tumours. Therefore, we propose that, when diagnosing an infiltrating malignant tumour, the percentage of tumour cells should be routinely recorded in the pathology reports. (AU)

A signature of 14 immune-related gene pairs predicts overall survival in gastric cancer

Objective Increasing evidence demonstrates that immune signature plays an important role in the prognosis of gastric cancer (GC). We aimed to develop and validate a robust immune-related gene pair (IRGP) signature for predicting the prognosis of GC patients. Methods RNA-Seq data and corresponding clinical information of GC cohort were downloaded from the TCGA (The Cancer Genome Atlas Program) data portal. GSE84437 and GSE15459 microarray datasets were included as independent external cohorts. Least absolute shrinkage and selection operator (LASSO) regression analysis was used to build the best prognostic signature. All patients were classified into the high immune-risk and low immune-risk groups via the optimal cut-off of the signature scores determined by time-dependent receiver-operating characteristic (ROC) curve analysis. The prognostic role of the signature was measured by a log-rank test and a Cox proportional hazard regression model. Results 14 immune gene pairs consisting of 25 unique genes were identified to construct the immune prognostic signature. High immune-risk groups showed poor prognosis in the TCGA datasets and GSE84437 datasets as well as in the GSE15459 datasets (all P < 0.001). The 14-IRGP signature was an independent prognostic factor of GC after adjusting for other clinical factors (P < 0.05). Functional analysis revealed that DNA integrity checkpoint, DNA replication, T-cell receptor signaling pathway, and B-cell receptor signaling pathway were enriched in the low immune-risk groups. B cells naive and Monocytes were significantly higher in the high-risk group, and B-cell memory and T-cell CD4 memory activated were significantly higher in the low-risk group. The prognostic signature based on IRGP reflected infiltration by several types of immune cells. Conclusion The novel proposed clinical-immune signature is a promising biomarker for prediction overall survival in patients with GC and providing new insights into the treatment strategies (AU)

Immunomodulatory effects of two silymarin isomers in a Balb/c mouse model of allergic asthma

INTRODUCTION AND OBJECTIVES: Allergic asthma is a complex chronic disease of the respiratory system presenting with cough, dyspnea, wheezing and airway obstruction. More than 300 million people of all age spectrums suffer from asthma worldwide. Immunological and inflammatory processes are main contributors to asthma. Cytokines produced by T helper 2 lymphocytes play main roles in asthma development and progression. Silymarin, a therapeutic agent with anti-oxidative properties, is a main component of Silybium marinum. We herein aimed to compare the anti-inflammatory and anti-allergic effects of two silymarin isomers, isosilybin A and silydianin, in the treatment of allergic asthma. MATERIALS AND METHODS: After isolating and purifying isosilybin A and silydianin, Balb/c mouse model of allergic asthma was produced using ovalbumin injection. Seventy mice were categorized into five (1 normal and 4 asthmatic) groups (n = 14 per group). Mice in three of four asthmatic groups were treated with either isosilybin A, silydianin or budesonide. The 4th asthmatic group was used as positive control, with the non-asthmatic group serving as negative control. Airway hyperresponsiveness (AHR) and levels of IL-4, IL-5 and IL-13 in the BAL fluid were determined. Gene expressions of IL-4, IL-5, IL-13 and MUC5ac, as well as IgE serum level were also measured. Cellular composition of BAL fluid and lungs histopathology were finally investigated. RESULTS: Isosilybin A and silydianin reduced eosinophilic infiltration of lungs, IL-4 and IL-5 levels in BAL fluid, IL-4 and IL-5 gene expressions, as well as AHR in Balb/c mouse model of asthma. However, no significant changes were observed in IL-13 level and mucus hyper-secretion. CONCLUSION: According to our study, isosilybin A and silydianin can control main symptoms of asthma by modulating immune responses

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Estudio de la expresión de factores óseos en el hueso murino ante la falta de pleiotrofina y sus cambios en la situación inflamatoria / Study of bone factor expression in murine model in the absence of pleiotrophin and its changes in the inflammatory situation

La pleitrofina (PTN) en un péptido implicado en el desarrollo y el mantenimiento del tejido óseo, y con importantes funciones en los procesos inflamatorios. Sin embargo, la deleción de la PTN en modelos murinos no produce un deterioro óseo significativo, sin que hasta la fecha se hayan estudiado los mecanismos que compensan su perdida. Con este estudio quisimos comprobar cómo afecta la deleción de PTN y la inflamación aguda a la expresión de factores óseos. Para ello empleamos ratones hembra de tres meses deficientes para la PTN (PTNKo) a las que indujimos una inflamación aguda por administración de lipopolisacárido (LPS). Se aislaron las vértebras y las tibias para poder medir la expresión de genes y realizar un recuento de osteocitos. En cultivos celulares comprobamos si la PTN podía proteger a células MC3T3 (osteoblásticas) y MLOY4 (osteocitos) de la inducción de muerte celular producida por etopósido. Nuestros resultados muestran que la expresión de osteocalcina está incrementada en las vértebras de los ratones PTNKo, y que la inflamación produjo el incremento de expresión de podaplanina (E11), conexina 43 (Cox43) y el péptido relacionado con la parathormona (PTHrP) en los ratones PTNKo tratados con LPS. La administración de PTN redujo de manera significativa la muerte inducida por etopósido en cultivos de células MC3T3 y MLOY4. En conclusión, la deficiencia de PTN indujo un aumento de la expresión de OCN, y la inflamación aguda produjo la sobrexpresión de E11, PTHrP, y Cox43 en ratones PTNKo. La PTN aumentó la viabilidad de células osteblásticas y osteocitos frente al tratamiento con etopósido

Pleiotrophin (PTN) is a peptide involved in the development and maintenance of bone tissue with important functions in inflammatory processes. However, the deletion of PTN in murine models does not produce a significant bone deterioration, but the mechanisms that compensate for its loss have not been studied to date. Our study was aimed at verifying how the deletion of PTN and acute inflammation affect the expression of bone factors. To this end, we used three-month-old female mice deficient for PTN (PTNKo) to which we induced acute inflammation by administration of lipopolysaccharide (LPS). Vertebrae and tibiae were isolated to measure gene expression and carry out an osteocyte count. In cell cultures, we checked whether PTN could protect MC3T3 (osteoblast) and MLOY4 (osteocyte) cells from the induction of cell death caused by etoposide. Our results show that the expression of osteocalcin is increased in the vertebrae of PTNKo mice, and that inflammation increased the expression of podhalanin (E11), connexin 43 (Cox43) and the parathormone-related peptide (PTHrP) in the PTNKo mice treated with LPS. Administering PTN significantly reduced etoposide-induced death in MC3T3 and MLOY4 cell cultures. Thus, PTN deficiency induced increased expression of OCN, and acute inflammation produced overexpression of E11, PTHrP, and Cox43 in PTNKo mice. PTN increased the viability of osteoblastic cells and osteocytes compared to etoposide treatment