Sildenafil normalizes MALAT1 level in diabetic cardiomyopathy

Introduction A large body of evidence recently highlighted the involvement of long non-coding RNAs (lncRNAs) in cardiovascular disease [1] and some dysregulated lncRNAs have been associated with diabetic cardiomyopathy [2\u20135]. Among them, a higher expression of the lncRNA metastasis associated lung adenocarcinoma transcript 1 (MALAT1) has been observed in diabetic cardiomyopathy [6, 7]. However, a clear understanding of the molecular mechanisms leading to pathological regulation of lncRNAs in diabetic cardiomyopathy is still missing. Our prior work by Barbati et al. [8], established that, in the presence of high glucose, nitric oxide (NO) signaling derangement might alter the epigenetic landscape of cardiac cells, both in vitro and in vivo, via transcription factor CREM activation. Aim The present study is aimed at investigating the role of high glucose (HG) and NO pathway in the regulation of MALAT1 in the heart of mice after 6 months of prolonged hyperglycemia and in two cellular models of cardiomyocytes exposed to HG

MALAT1 as a Regulator of the Androgen-Dependent Choline Kinase A Gene in the Metabolic Rewiring of Prostate Cancer

Simple Summary Despite the rapid advance in cancer therapies, treatment-resistant relapse remains a significant challenge in cancer treatment. Acquired resistance arises during or after treatment administration, and is usually the main contributor to relapse. For example, prostate cancer, the most frequent type of cancer in the elderly male population, frequently develops into aggressive forms resistant to chemical and hormonal therapies. In this condition, the so-called "cholinic phenotype" that is characterized by the overexpression of choline kinase alpha (CHKA) and increased phosphocholine levels leads to aberrant lipid metabolism. Our work demonstrates that CHKA, which is necessary for membrane phospholipid synthesis, is a target of the long non-coding RNA MALAT1. This study helps to further decipher how MALAT1 affects the regulation of crucial phospholipid/sphingolipid metabolic enzymes, as well as how the androgen receptor pathway is involved in MALAT1-dependent transcriptional regulation. Background. Choline kinase alpha (CHKA), an essential gene in phospholipid metabolism, is among the modulated MALAT1-targeted transcripts in advanced and metastatic prostate cancer (PCa). Methods. We analyzed CHKA mRNA by qPCR upon MALAT1 targeting in PCa cells, which is characterized by high dose-responsiveness to the androgen receptor (AR) and its variants. Metabolome analysis of MALAT1-depleted cells was performed by quantitative High-resolution 1 H-Nuclear Magnetic Resonance (NMR) spectroscopy. In addition, CHKA genomic regions were evaluated by chromatin immunoprecipitation (ChIP) in order to assess MALAT1-dependent histone-tail modifications and AR recruitment. Results. In MALAT1-depleted cells, the decrease of CHKA gene expression was associated with reduced total choline-containing metabolites compared to controls, particularly phosphocholine (PCho). Upon MALAT1 targeting a significant increase in repressive histone modifications was observed at the CHKA intron-2, encompassing relevant AR binding sites. Combining of MALAT1 targeting with androgen treatment prevented MALAT1-dependent CHKA silencing in androgen-responsive (LNCaP) cells, while it did not in hormone-refractory cells (22RV1 cells). Moreover, AR nuclear translocation and its activation were detected by confocal microscopy analysis and ChIP upon MALAT1 targeting or androgen treatment. Conclusions. These findings support the role of MALAT1 as a CHKA activator through putative association with the liganded or unliganded AR, unveiling its targeting as a therapeutic option from a metabolic rewiring perspective

MALAT1 and HOTAIR long non-coding RNAs play opposite role in estrogen-mediated transcriptional regulation in prostate cancer cells

In the complex network of nuclear hormone receptors, the long non-coding RNAs (lncRNAs) are emerging as critical determinants of hormone action. Here we investigated the involvement of selected cancer-associated lncRNAs in Estrogen Receptor (ER) signaling. Prior studies by Chromatin Immunoprecipitation (ChIP) Sequencing showed that in prostate cancer cells ERs form a complex with the endothelial nitric oxide synthase (eNOS) and that in turn these complexes associate with chromatin in an estrogen-dependent fashion. Among these associations (peaks) we focused our attention on those proximal to the regulatory region of HOTAIR and MALAT1. These transcripts appeared regulated by estrogens and able to control ERs function by interacting with ER\u3b1/ER\u3b2 as indicated by RNA-ChIP. Further studies performed by ChIRP revealed that in unstimulated condition, HOTAIR and MALAT1 were present on pS2, hTERT and HOTAIR promoters at the ERE/eNOS peaks. Interestingly, upon treatment with17\u3b2-estradiol HOTAIR recruitment to chromatin increased significantly while that of MALAT1 was reduced, suggesting an opposite regulation and function for these lncRNAs. Similar results were obtained in cells and in an ex vivo prostate organotypic slice cultures. Overall, our data provide evidence of a crosstalk between lncRNAs, estrogens and estrogen receptors in prostate cancer with important consequences on gene expression regulatio

Neutralizing antibodies to Omicron after the fourth SARS-CoV-2 mRNA vaccine dose in immunocompromised patients highlight the need of additional boosters

IntroductionImmunocompromised patients have been shown to have an impaired immune response to COVID-19 vaccines.MethodsHere we compared the B-cell, T-cell and neutralizing antibody response to WT and Omicron BA.2 SARS-CoV-2 virus after the fourth dose of mRNA COVID-19 vaccines in patients with hematological malignancies (HM, n=71), solid tumors (ST, n=39) and immune-rheumatological (IR, n=25) diseases. The humoral and T-cell responses to SARS-CoV-2 vaccination were analyzed by quantifying the anti-RBD antibodies, their neutralization activity and the IFN-γ released after spike specific stimulation.ResultsWe show that the T-cell response is similarly boosted by the fourth dose across the different subgroups, while the antibody response is improved only in patients not receiving B-cell targeted therapies, independent on the pathology. However, 9% of patients with anti-RBD antibodies did not have neutralizing antibodies to either virus variants, while an additional 5.7% did not have neutralizing antibodies to Omicron BA.2, making these patients particularly vulnerable to SARS-CoV-2 infection. The increment of neutralizing antibodies was very similar towards Omicron BA.2 and WT virus after the third or fourth dose of vaccine, suggesting that there is no preferential skewing towards either virus variant with the booster dose. The only limited step is the amount of antibodies that are elicited after vaccination, thus increasing the probability of developing neutralizing antibodies to both variants of virus.DiscussionThese data support the recommendation of additional booster doses in frail patients to enhance the development of a B-cell response directed against Omicron and/or to enhance the T-cell response in patients treated with anti-CD20

Efecto del cierre de las escuelas sobre el brote de influenza A H1N1 en Tierra del Fuego, Argentina Impact of school closings on the influenza A (H1N1) outbreak in Tierra del Fuego, Argentina

Se evaluó el efecto del cierre de las escuelas en la reducción del número de casos de enfermedades del tipo influenza durante un brote de influenza A por el virus H1N1, de características pandémicas, en las dos principales ciudades de la provincia de Tierra del Fuego, en la región austral de Argentina, en conjunto con otras medidas de control. Se comparó la incidencia antes y después del cierre de las escuelas en 2009 mediante la prueba de la t para muestras relacionadas. Hasta la semana 40 se detectaron 6 901 casos de enfermedades del tipo influenza, de ellos 281 se confirmaron como influenza A H1N1 mediante pruebas de laboratorio; 38 pacientes recibieron hospitalización. Se observó una reducción de cerca de 10 veces en la incidencia promedio registrada en los centros de salud después de la intervención. Los resultados indican que el cierre de las escuelas durante el brote de influenza A H1N1 se acompañó de una reducción significativa de la incidencia de enfermedades tipo influenza, aunque no se debe descartar el efecto de otras acciones, como el tratamiento de los casos y la profilaxis de sus contactos. Esta intervención, aplicada oportunamente, junto con otras medidas, puede contribuir a reducir la propagación de los brotes de influenza."The impact of school closings on reducing the number of cases of influenza-like illness during an outbreak of influenza A (H1N1), which reached pandemic proportions, was assessed, along with other control measures, in the two main cities of Tierra del Fuego Province in southern Argentina. The incidence before and after the school closings in 2009 was compared by means of the t-test for related samples. By week 40, 6 901 cases of influenza-like illness had been detected, 281 of which were confirmed as influenza A (H1N1) through laboratory tests; 38 patients were hospitalized. After the intervention, there were nearly 10 times fewer cases than the average recorded in the health centers. The results indicate that closing schools during the influenza A (H1N1) outbreak resulted in a significantly lower incidence of influenza-like illness. However, the impact of other measures, such as case management and protection against exposure, should not be ignored. Timely implementation of this intervention, together with other measures, can help minimize the spread of influenza outbreaks

Genetic parameters estimation in quantitative traits of a cross of triticale (x Triticosecale W.)

Triticale (x Triticosecale Wittmack) is a cereal used in Argentina as a winter seasonal pasture or feeding grain. Efforts made on breeding have led to varieties with potential grain yield similar or greater than with wheat. The aim of this study was to determine the gene effects, heritability, genetic gain, heterosis and inbreeding depression in various quantitative traits of a cross from contrasting lines of triticale using generation means analysis. The thousand kernel weight was adjusted to a model of additive-dominance, while the rest of the traits exhibited significant epistatic effects: additive x additive (i) for hectoliter weight, additive x dominance (j) for number grains per spike and grain yield per plant. Other traits showed a more complex as inheritance additive x additive (i) and additive x dominance (j) interactions effects for spike length or additive x dominance (j) and dominance x dominance (l) interactions effects for number of spikes per plant and number of spikelets per spike. High values of heritability and genetic gain were also detected for grain yield, spikelets per spike, spike per plant and hectoliter weight. All traits except spike per plant had positive relative mid-parent heterosis, but none resulted in positive heterobeltiosis, therefore this cross is not recommended for obtaining hybrids varieties. Selection in early generations by thousand kernel weight and hectoliter weight would be efficient, while the selection in advanced generations where segregation is minimal would be recommended for the remaining traits

First episode psychoses in people over-35 years old: uncovering potential actionable targets for early intervention services

The traditional youth-oriented design of Early Intervention Services (EIS) may lead to the exclusion of patients who have their psychotic onset later in life. A retrospective study was conducted to compare first-episode psychosis (FEP) patients who accessed treatment when aged ≤ 35 years with those ≥36+. A total of 854 patients were identified among 46,222 individuals who had access to community psychiatric services from 1991 to 2021. FEP were aged 18–65, received care between 2012 and 2021 and had a diagnosis of affective or non-affective FEP. Two groups were identified (FEP diagnosed at age ≤ 35 vs ≥ 36) and compared for sociodemographic and clinical characteristics. Most patients were diagnosed when aged ≥ 36+ (61.8%). Compared to the ≤ 35 group, older patients were more likely to be women, married and diagnosed with affective psychosis, and they were less frequently hospitalized. Long-acting injectables antipsychotics (LAI) were less frequently prescribed in the ≥ 36+ group, whereas antidepressants were more frequently prescribed compared to those aged ≤ 35. In both age groups, women were less frequently prescribed LAIs compared to men. These findings highlight the need to reorient EIS to accommodate the needs of older FEP, especially women

H19-Dependent Transcriptional Regulation of β3 and β4 Integrins Upon Estrogen and Hypoxia Favors Metastatic Potential in Prostate Cancer

Estrogen and hypoxia promote an aggressive phenotype in prostate cancer (PCa), driving transcription of progression-associated genes. Here, we molecularly dissect the contribution of long non-coding RNA H19 to PCa metastatic potential under combined stimuli, a topic largely uncovered. The effects of estrogen and hypoxia on H19 and cell adhesion molecules’ expression were investigated in PCa cells and PCa-derived organotypic slice cultures (OSCs) by qPCR and Western blot. The molecular mechanism was addressed by chromatin immunoprecipitations, overexpression, and silencing assays. PCa cells’ metastatic potential was analyzed by in vitro cell-cell adhesion, motility test, and trans-well invasion assay. We found that combined treatment caused a significant H19 down-regulation as compared with hypoxia. In turn, H19 acts as a transcriptional repressor of cell adhesion molecules, as revealed by up-regulation of both β3 and β4 integrins and E-cadherin upon H19 silencing or combined treatment. Importantly, H19 down-regulation and β integrins induction were also observed in treated OSCs. Combined treatment increased both cell motility and invasion of PCa cells. Lastly, reduction of β integrins and invasion was achieved through epigenetic modulation of H19-dependent transcription. Our study revealed that estrogen and hypoxia transcriptionally regulate, via H19, cell adhesion molecules redirecting metastatic dissemination from EMT to a β integrin-mediated invasion

Transcription factor CREM mediates high glucose response in cardiomyocytes and in a male mouse model of prolonged hyperglycemia

This study aims at investigating the epigenetic landscape of cardiomyocytes exposed to long elevated glucose levels in vitro and in vivo Exposure of mouse HL-1 and rat differentiated H9C2 cardiomyocyte cell lines to high glucose (30 mM) for 72 hours determined some epigenetic changes including upregulation of class I and III HDAC protein levels and activity, inhibition of histone acetylase p300 activity, increase in histone H3 lysine (K) 27 trimethylation (H3K27me3) and reduction in H3K9 acetylation (H3K9Ac). Gene expression analysis focused on cardiotoxicity, revealed that high glucose induced markers associated with tissue damage, fibrosis and cardiac remodelling such as Nexilin, Versican, CREM and ADRA2. Notably, the transcription factor CREM was found to be important in the regulation of cardiotoxicity-associated genes as assessed by specific siRNA and chromatin immunoprecipitation experiments. In CD1 mice, made hyperglycaemic by STZ injection, cardiac structural alterations were evident at six months after STZ treatment and were associated with a significant increase of H3K27me3 and reduction of H3K9Ac. Consistently, Nexilin, CREM and ADRA2 expression was significantly induced at RNA and protein level. Confocal microscopy analysis of Nexilin localization showed this protein irregularly distributed along the sarcomeres in the heart of hyperglycaemic mice. This evidence suggested for a structural alteration of cardiac Z-disk with potential consequences on contractility. In conclusion, high glucose may alter the epigenetic landscape of cardiac cells. Sildenafil, restoring cGMP levels, counteracted the increase of CREM and Nexilin providing a protective effect in the presence of hyperglycemia

Establishment of a protocol to extend the lifespan of human hormone-secreting pituitary adenoma cells

Purpose The aim of this study was to generate immortalized human anterior pituitary adenoma cells. Reliable cell models for the study of human pituitary adenomas are as yet lacking and studies performed so far used repeated passaging of freshly excised adenomas, with the attendant limitations due to limited survival in culture, early senescence, and poor reproducibility. Methods & Results We devised a technique based upon repeated co-transfections of two retroviral vectors, one carrying the catalytic subunit of human telomerase, hTERT, the other SV40 large T antigen. This approach extended the lifespan of cells derived from a human growth hormone-secreting adenoma up to 18 months while retaining morphology of primary cells, growth hormone synthesis and growth hormone secretion. Conclusions Our attempt represents the first demonstration of successful lifespan extension of human growth hormone-secreting pituitary adenoma cells via co-transfection of hTERT and SV40T and paves the way to future attempts to obtain stable cell lines