Oral pathology and oral medicine in Latin American countries: current stage.

BACKGROUND: Oral Pathology (OP) and Oral Medicine (OM) are specialties in dentistry whose main objective is the diagnosis and treatment of oral and maxillofacial diseases, and aspects related to the academic training of professionals and fields of practice are distinct and heterogeneous around the world. This study aimed to evaluate professional training and areas of activity in OP and OM in Latin American countries. MATERIAL AND METHODS: A questionnaire was sent to 11 countries, with a professional in each country responsible for answering it. The questionnaire had 21 questions related to the process of professional training, areas of practice, the existence of scientific events in each country, and also collected demographic and population information. RESULTS: OP and OM are practiced in all the countries studied, but the specialty is not recognized in all of them. Brazil was the first to recognize both as a specialty. Postgraduate programs designed to train specialists are available in various countries. Two countries offer residency programs, 6 countries provide specialization courses, 6 offer master's programs, and 3 have doctoral programs. Brazil boasts the highest number of undergraduate courses (n=412), while Uruguay has the lowest (n=2). Professional societies representing the specialty exist in ten countries. Brazil has the highest number of OP and OM specialists (n=422 and 1,072), while Paraguay has the smallest number (n=1 and 3). CONCLUSIONS: Although both specialties are widely practiced around the globe, professional training, the number of dentists trained and the fields of professional practice are very different between the countries studied.

G-protein coupled receptor 5C (GPRC5C) is required for osteoblast differentiation and responds to EZH2 inhibition and multiple osteogenic signals.

Osteoblast differentiation is epigenetically suppressed by the H3K27 methyltransferase EZH2, and induced by the morphogen BMP2 and transcription factor RUNX2. These factors also regulate distinct G protein coupled receptors (GPRCs; e.g., PTH1R, GPR30/GPER1). Because GPRCs transduce many physiological stimuli, we examined whether BMP2 or EZH2 inhibition (i.e., GSK126) regulates other GPRC genes in osteoblasts. RNA-seq screening of >400 mouse GPRC-related genes showed that many GPRCs are downregulated during osteogenic differentiation. The orphan receptor GPRC5C, along with a small subset of other GPRCs, is induced by BMP2 or GSK126 during Vitamin C dependent osteoblast differentiation, but not by all-trans retinoic acid. ChIP-seq analysis revealed that GSK126 reduces H3K27me3 levels at the GPRC5C gene locus in differentiating MC3T3-E1 osteoblasts, consistent with enhanced GPRC5C mRNA expression. Loss of function analyses revealed that shRNA-mediated depletion of GPRC5C decreases expression of bone markers (e.g., BGLAP and IBSP) and mineral deposition in response to BMP2 or GSK126. GPRC5C mRNA was found to be reduced in the osteopenic bones of KLF10 null mice which have compromised BMP2 signaling. GPRC5C mRNA is induced by the bone-anabolic activity of 17ß-estradiol in trabecular but not cortical bone following ovariectomy. Collectively, these findings suggest that GPRC5C protein is a key node in a pro-osteogenic axis that is normally suppressed by EZH2-mediated H3K27me3 marks and induced during osteoblast differentiation by GSK126, BMP2, and/or 17ß-estradiol. Because GPRC5C protein is an understudied orphan receptor required for osteoblast differentiation, identification of ligands that induce GPRC5C signaling may support therapeutic strategies to mitigate bone-related disorders.

The epigenetic regulator BRD4 is required for myofibroblast differentiation of knee fibroblasts.

Arthrofibrosis, which is characterized by excessive scar tissue and limited motion, can complicate the daily functioning of patients after total knee arthroplasty (TKA). Molecular hallmarks of arthrofibrosis include pathologic accumulation of myofibroblasts and disproportionate collagen deposition. Epigenetic mechanisms, including posttranslation modification of histones, control gene expression and may regulate fibrotic events. This study assessed the role of the bromodomain and extra-terminal (BET) proteins on myofibroblast differentiation. This group of epigenetic regulators recognize acetylated lysines and are targeted by a class of drugs known as BET inhibitors. RNA-seq analysis revealed robust mRNA expression of three BET members (BRD2, BRD3, and BRD4) while the fourth member (BRDT) is not expressed in primary TKA knee outgrowth fibroblasts. RT-qPCR and western blot analyses revealed that BET inhibition with the small molecule JQ1 impairs TGFß1-induced expression of ACTA2, a key myofibroblast marker, in primary outgrowth knee fibroblasts. Similarly, JQ1 administration also reduced COL3A1 mRNA levels and collagen deposition as monitored by picrosirius red staining. Interestingly, the inhibitory effects of JQ1 on ACTA2 mRNA and protein expression, as well as COL3A1 expression and collagen deposition, were paralleled by siRNA-mediated depletion of BRD4. Together, these data reveal that BRD4-mediated epigenetic events support TGFß1-mediated myofibroblast differentiation and collagen deposition as seen in arthrofibrosis. To our knowledge, these are the first studies that assess epigenetic regulators and their downstream events in the context of arthrofibrosis. Future studies may reveal clinical utility for drugs that target epigenetic pathways, specifically BET proteins, in the prevention and treatment of arthrofibrosis.

Brd4 is required for chondrocyte differentiation and endochondral ossification.

Differentiation of multi-potent mesenchymal stromal cells (MSCs) is directed by the activities of lineage-specific transcription factors and co-factors. A subset of these proteins controls the accessibility of chromatin by recruiting histone acetyl transferases or deacetylases that regulate acetylation of the N-termini of H3 and H4 histone proteins. Bromodomain (BRD) proteins recognize these acetylation marks and recruit the RNA pol II containing transcriptional machinery. Our previous studies have shown that Brd4 is required for osteoblast differentiation in vitro. Here, we investigated the role of Brd4 on endochondral ossification in C57BL/6 mice and chondrogenic differentiation in cell culture models. Conditional loss of Brd4 in the mesenchyme (Brd4 cKO, Brd4fl/fl: Prrx1-Cre) yields smaller mice that exhibit alteration in endochondral ossification. Importantly, abnormal growth plate morphology and delayed long bone formation is observed in juvenile Brd4 cKO mice. One week old Brd4 cKO mice have reduced proliferative and hypertrophic zones within the physis and exhibit a delay in the formation of the secondary ossification center. At the cellular level, Brd4 function is required for chondrogenic differentiation and maturation of both ATDC5 cells and immature mouse articular chondrocytes. Mechanistically, Brd4 loss suppresses Sox9 levels and reduces expression of Sox9 and Runx2 responsive endochondral genes (e.g., Col2a1, Acan, Mmp13 and Sp7/Osx). Collectively, our results indicate that Brd4 is a key epigenetic regulator required for normal chondrogenesis and endochondral ossification.

Brd4 Inactivation Increases Adenoviral Delivery of BMP2 for Paracrine Stimulation of Osteogenic Differentiation as a Gene Therapeutic Concept to Enhance Bone Healing.

Bromodomain (BRD) proteins are histone code interpreters that recognize acetylated lysines and link the dynamic state of chromatin with the transcriptional machinery. Here, we demonstrate that ablation of the Brd4 gene in primary mouse bone marrow-derived mesenchymal stem cells via a conditional Brd4fl/fl allele suppresses osteogenic lineage commitment. Remarkably, loss of Brd4 function also enhances expression of genes in engineered adenoviral vectors, including Cre recombinase and green fluorescent protein (GFP). Similarly, vector-based expression of BMP2 mRNA and protein levels are enhanced upon Brd4 depletion in cells transduced with an adenoviral vector that expresses BMP2 (Ad-BMP2). Importantly, Brd4 depletion in MC3T3-E1 and human adipose-derived mesenchymal stem cells (AMSCs) transduced with Ad-BMP2 enhances osteogenic differentiation of naïve MC3T3-E1 cells via paracrine mechanisms based on transwell and conditioned medium studies. Our studies indicate that Brd4 depletion enhances adenoviral transgene expression in mammalian cells, which can be leveraged as a therapeutic strategy to improve viral vector-based gene therapies. © 2021 The Authors. JBMR Plus published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research.

Results of Shouldice hernia repair after 18 years of follow-up in all the patients.

PURPOSE: Evaluate the long-term efficacy of the Shouldice technique performed by non-specialized surgeons and also to reflex on the quality parameters necessary to safely assess hernia recurrence rates. METHODS: During 3 years, a prospective study was conducted in 243 adult men who underwent surgery for primary inguinal hernias by 13 junior surgeons with an interest in hernia surgery. Using local anesthesia, a classic 4 step Shouldice repair, with polypropylene or polyester, was performed. All patients were followed for 18 years. The follow-up met the nine quality criteria proposed by the authors. RESULTS: At 18 years, 80.2% of patients were followed and only 6.5% were lost. There were 7 recurrences in the first 10 years, 5 of them secondary to a direct hernia, and the same after 18 years. The recurrence rate was 2.88%. Tolerance of the local anesthesia was excellent in 91.4%of patients and, after 3 years, the pain was considered moderate or severe in 4 patients (1.8%). CONCLUSIONS: It is necessary to incorporate more demanding criteria in the assessment of recurrence, to give more valid results. The Shouldice technique remains a useful technique today not only in patients under 30 years of age, and in the absence of risk factors, but also in cases of intolerance, patient rejection or absence of mesh. In addition, it provides the clinical and economic advantages of being possible to perform it under local anesthesia.

Multiple pharmacological inhibitors targeting the epigenetic suppressor enhancer of zeste homolog 2 (Ezh2) accelerate osteoblast differentiation.

Skeletal development and bone formation are regulated by epigenetic mechanisms that either repress or enhance osteogenic commitment of mesenchymal stromal/stem cells and osteoblasts. The transcriptional suppressive trimethylation of histone 3 lysine 27 (H3K27me3) hinders differentiation of pre-committed osteoblasts. Osteoblast maturation can be stimulated by genetic loss of the H3K27 methyltransferase Ezh2 which can also be mimicked pharmacologically using the classical Ezh2 inhibitor GSK126. Identification of other Ezh2 inhibitors (iEzh2) that enhance osteogenic potential would increase chemical options for developing new bone stimulatory compounds. In this study, we examined a panel of iEzh2s and show that all eight inhibitors we tested are capable of accelerating osteoblast differentiation to different degrees at concentrations that are well below cytotoxic concentrations. Inhibition of Ezh2 is commensurate with loss of cellular H3K27me3 levels while forced expression of Ezh2 reverses the effect of Ezh2 suppression. Reduced Ezh2 function by siRNA depletion of Ezh2 mRNA and protein levels also stimulates osteoblastogenesis, consistent with the specificity of iEzh2 to target the active site of Ezh2. Diminished Ezh2 levels preempt the effects of iEzh2s on H3K27me3. GSK126, EPZ-6438 and siRNA depletion of Ezh2 each are effective in reducing H3K27me3 levels. However, EPZ-6438 is more potent than GSK126 in stimulating osteoblastogenesis, as reflected by increased extracellular matrix mineralization. Collectively, our data indicate that Ezh2 inhibitors properly target Ezh2 consistent with their biochemical affinities. The range of compounds capable of promoting osteogenesis presented in this study offers the opportunity to develop diverse bone anabolic strategies for distinct clinical scenarios, including spine fusion, non-union of bone and dental implant enhancement.

Effect of a Multicomponent Exercise Program (VIVIFRAIL) on Functional Capacity in Elderly Ambulatory: A Non-Randomized Clinical Trial in Mexican Women with Dynapenia.

BACKGROUND: During aging, loss of muscle strength (dynapenia) combined with unhealthy lifestyles and sedentarism can lead to functional limitations and dependency; currently there is still limited evidence about multicomponent training as a treatment for dynapenia and dependence in ambulatory older women. OBJECTIVE: To evaluate the effect of a multicomponent physical exercise program (VIVIFRAIL) on physical functionality in elderly ambulatory women with dynapenia. DESIGN, SETTING AND PARTICIPANTS: A non-randomized clinical trial was realized in 61 ambulatory older women (65-80 years old) with dynapenia that belonged from two Integral Gerontological Centers (IGC) of Hidalgo, Mexico, from June to December 2019. INTERVENTION: The control group (CG) received the physical daily training applied in the IGC (yoga, cardio-dance or tai chi) and the intervention group (IG) participated in a multicomponent program training (resistance, flexibility, balance and gait) called "VIVIFRAIL" for 12 weeks, minimum 3 weekly sessions of 45-60 minutes. MEASURES: All participants were evaluated at baseline (0 weeks), intermediate (6 weeks) and final (12 weeks), evaluations included glucose, blood pressure (SBP and DBP), anthropometric test, body composition evaluation and functional performance test with Short Physical Performance Battery (SPPB), Timed Up and Go Test (TUGT), muscle strength and a falls risk short test. RESULTS: 52 older women completed the study, mean age was 71.9 ± 4.46. According to SPPB passport classification was 1 Type A, 1 Type B+, 9 Type C, 7 Type C+ and 9 Type D. After 12 weeks of intervention, statistical analysis showed that multicomponent exercise significantly improved strength (p=<0.001), gait speed (p<0.001), standing from a chair (p<0.001) and TUGT (p<0.001). About falls risk, intervention group went from 70% to 12.5% while control group went from 52.4% to 63.6% (p<0.05). CONCLUSION: Older women who completed the 12 weeks multicomponent exercise program significantly improved their functionality in muscle strength, gait speed (3mt and 6mt), standing from a chair and TUGT tests. Also, it showed a significantly decrease in falls risk, therefore, this type of intervention can reduce the risk of frailty in the elderly.

Differences in Cytotoxicity of Lidocaine, Ropivacaine, and Bupivacaine on the Viability and Metabolic Activity of Human Adipose-Derived Mesenchymal Stem Cells.

PURPOSE: We evaluated biological effects of distinct local anesthetics on human adipose-derived mesenchymal stem cells when applied to reduce periprocedural pain during mesenchymal stem cell injections. METHODS AND MATERIALS: Metabolic activity (MTS assay), viability (Live/Dead stain), and gene expression (quantitative real-time reverse-transcriptase polymerase chain reaction) were measured in mesenchymal stem cells incubated with various concentrations of lidocaine, ropivacaine, or bupivacaine during a 12-hr time course. RESULTS: Cell viability and metabolic activity decreased in a dose, time, and substance-specific manner after exposure to lidocaine, ropivacaine, and bupivacaine, with ropivacaine being the least cytotoxic. Cell viability decreases after brief exposure (<1.5 hrs) at clinically relevant concentrations (eg, 8 mg/ml of lidocaine, 2.5 mg/ml of ropivacaine or bupivacaine). Mesenchymal stem cells exposed to local anesthetics change their expression of mRNA biomarkers for stress response (EGR1, EGR2), proliferation (MKI67, HIST2H4A), ECM (COL1A1, COL3A1), and cell surface marker (CD105). CONCLUSIONS: Local anesthetics are cytotoxic to clinical-grade human mesenchymal stem cells in a dose-, time-, and agent-dependent manner and change expression of ECM, proliferation, and cell surface markers. Lidocaine and bupivacaine are more cytotoxic than ropivacaine. Single-dose injections of local anesthetics may affect the biological properties of mesenchymal stem cells in vitro but may not affect the effective dose of MSCs in a clinical setting.

Terapias de reemplazo renal contínuas (TRRC) en pacientes críticos inmunosuprimidos / Continuous renal replacement therapies (CRRT) in critically ill immunosuppressed patients

La injuria renal aguda (IRA) se caracteriza por un abrupto deterioro de la función renal asociado a lteraciones hidroelectrolíticas y metabólicas. La misma es frecuente en la unidad de cuidados intensivos (UCI) pediátricos y tiene un impacto significativo en la morbilidad y mortalidad. Las principales indicaciones de terapia de reemplazo renal (TRR) incluyen la corrección de los trastornos metabólicos y el manejo de la sobrecarga de fluidos. Varios modos de TRR pueden ser utilizadas en la UCI: hemodiálisis intermitente, diálisis peritoneal y las terapias de reemplazo renal continuas (TRRC). Las terapias de reemplazo renal continuas han ganado un rol preponderante en Cuidados Críticos ya que posibilitan dializar a pacientes hemodinámicamente inestables. Del total de pacientes admitidos en la UCI (n:1506) desde enero 2012 hasta diciembre 2018, requirieron TRRC el 6,7% (n: 102). La mortalidad predicha por el Score PIM3 fue de 19,53%, la mediana de edad en meses fue de 60 (RIC 25-75: 12-144), no hubo diferencias en cuanto al sexo. Los diagnósticos más frecuentes fueron trasplantados de órganos sólidos 33%, seguidos de trasplante de células progenitoras hematopoyéticas (TCPH) el 26%. La mediana de los días de internación fue de 16 (RIC 25-75: 7-29) y de días de requerimiento de una TRRC 5 (RIC 25-75 3-9). La técnica dialítica más utilizada fue CVVHD, en el 87% de los pacientes. La mortalidad global fue del 75%, presentando los pacientes con TCPH mayor mortalidad con respecto a otros diagnósticos. Se debe reconocer y categorizar precozmente a los pacientes con mayor riesgo de desarrollar IRA y aplicar medidas de nefroprotección para mejorar su sobrevida (AU)

Acute renal injury (IRA) is characterized by sudden deterioration of kidney function associated with hydroelectrolytic and metabolic disturbances. IRA is common in the pediatric intensive care unit (ICU) and has a significant impact on morbidity and mortality. The main indications for renal replacement therapy (RRT) include correction of the metabolic disorders and management of fluid overload. Different types of RRT may be used in the ICU: intermittent hemodialysis, peritoneal dialysis, and continuous renal replacement therapies (CRRT). Continuous renal replacement therapies have gained a major role in critical care as they allow for dialysis in hemodynamically unstable patients. Of all patients admitted to the ICU (n:1506) between January 2012 and December 2018, 6.7% required CRRT (n: 102). Predicted mortality rate according to the PIM3 score was 19.53%. Median age was 60 months (IQR 25-75: 12-144). No differences in sex were observed. The most common diagnoses were solid organ transplantation in 33%, followed by hematopoietic stem cell transplantation (HSCT) in 26%. Median length of hospital stay was 16 days (IQR 25-75: 7-29) and median days on CTTT was 5 (IQR 25-75 3-9). The most common dialysis technique was CVVHD, used in 87% of the patients. Overall mortality rate was 75%, with a higher mortality in HSCT patients compared to others. Patients at a higher risk of developing IRA should be timely recognized and categorized and nephroprotective measures should be started early to improve survival (AU)

Urgencias hematooncológicas: desde la perspectiva del intensivista / Hematology/oncology urgencies: from the perspective of the intensivist

Entre los años 2000 y 2016 en Argentina, se reportaron al Registro Oncopediátrico Hospitalario Argentino (ROHA) 22.450 casos de cáncer en niños menores de 15 años de edad. Las Leucemias constituyen la enfermedad oncológica más frecuente, seguida de los Tumores de Sistema Nervioso Central y los Linfomas. Esta distribución es similar a la descripta en los países desarrollados de Europa y Norteamérica. Su tasa de curación a nivel mundial, llega al 80% debido al uso de quimioterapia intensiva, situación que mejora la supervivencia pero que también aumenta la frecuencia de complicaciones. Estas complicaciones pueden ser debidas tanto al propio cáncer como al tratamiento y en ocasiones ser la primera manifestación de la enfermedad oncológica. Los eventos que amenazan la vida en pacientes inmunocomprometidos son mayores que en la población general, y cuando ocurren tienen una mortalidad elevada. El reconocimiento temprano es clave para el resultado en términos de sobrevida y disminución de la mortalidad. Las acciones deberán centrarse al reconocimiento temprano de eventos críticos en pacientes oncológicos. Los pacientes Hemato-Oncológicos constituyen un gran número de ingresos no planificados a las unidades de cuidados intensivos. Uno de cada 4 pacientes requerirá durante su evolución ingreso a Unidades de Cuidados Intensivos. El propósito de este artículo es describir tres de las urgencias oncológicas que requieren con mayor frecuencia admisión en UCI: la presentación y manejo del shock séptico, Shock Cardiogénico y las complicaciones neurológicas en los pacientes con leucemias agudas (AU)

Between 2000 and 2016, 22,450 cases of cancer in children younger than 15 years of age were reported to the Argentine Hospital Registry of Childhood Cancer (ROHA). Leukemia was the most common cancer reported, followed by central nervous system tumors and lymphoma. This distribution is similar to that described in the developed countries of Europe and North America. The worldwide cure rate is up to 80% due to the use of intensive chemotherapy, which improves survival but also increases the complication rate. These complications may be due both to the cancer itself and to the treatment and are sometimes the first manifestation of the disease. Life-threatening events are more common in immunocompromised patients than in the general population, and when they occur, the mortality rate is high. Early recognition is essential for the outcome in terms of survival and decreased mortality. Interventions should focus on early recognition of critical events in cancer patients. Patients with hematology-oncology diseases account for a large number of unplanned admissions to intensive care units (ICU), while one in four of these patients will require admission to the ICU in the course of their disease. The aim of this study was to describe three oncology emergencies that most frequently require ICU admission: septic shock and its management, cardiogenic shock, and neurological complications in patients with acute leukemia (AU)

Prospecting bacterial consortia from a geothermal site for metals biotransformation.

Biomats that flourished in a fumarole located on the geothermal site Los Azufres (Mexico) were used as inocula to select aerobic and sulfate-reducing bacteria consortia for studying their capacity to reduce hexavalent chromium [Cr(VI)], aiming to use these consortia in biotransformation technologies. The sample site is characterized by slightly warm (nearly 27 [Formula: see text]C), acid (pH 3) and about hypoxic (1.8 mg L[Formula: see text] of dissolved oxygen) conditions. Four culture systems (2 aerobic and 2 anaerobic) were investigated, including their enzymatic activity, capacity to produce biofilms, and an analysis of the total bacterial populations. For the anaerobic condition (using sulfate and sulfur as electron acceptors), four pH values (from 2 to 8) and four carbon sources (pyruvate, glycerol, Na-lactate and Na-acetate) were probed. Significant biological Cr(VI) removal was observed for all the pH values probed, particularly during the first 12 h, being more effective at the most acid conditions. At a pH value of 4 and using pyruvate as carbon source, 100 mg L[Formula: see text] of Cr(VI) were completely depleted in less than 12 h, while the use of Na-lactate was less effective but still reasonable. These results indicate that sulfate-reducing bacteria consortia from geothermal sites like the one studied here are capable of biotransforming Cr(VI) and have the potential to provide metal bioremediation technologies.

Differential biomarker profiles between unprovoked venous thromboembolism and cancer.

BACKGROUND: The relationship between cancer and venous thromboembolic disease (VTD) are complex because the activated coagulation factors are not only involved in thrombosis but also in malignant processes, such as angiogenesis and metastasis. OBJECTIVE: To compare phenotypes of extracellular vesicles (EVs), and levels of D-dimer, soluble P-selectin (sP-selectin) and antigenic tissue factor (TF) between unprovoked VTD patients, who did not develop cancer during one-year follow-up, and those with advanced stage of cancer but not associated with VTD. METHODS: A prospective study in which we included 138 unprovoked VTD patients and 67 advanced cancer patients, who did not develop thrombosis. Levels of EVs of different cellular origin (platelet, endothelium and leukocyte), EVs positive for tissue factor (TF) and P-selectin glycoprotein ligand 1 were quantified by flow cytometry. D-dimer, soluble P-selectin (sP-selectin) and antigenic TF were determined by ELISA. RESULTS: TF-positive EVs, D-dimer, and sP-selectin were markedly elevated in unprovoked VTD patients compared to cancer patients without association with thrombosis. CONCLUSIONS: Levels of TF-positive EVs, D-dimer and sP-selectin are able to discriminate between unprovoked VTD patients not related to cancer and cancer patients not associated with VTD. These results could lead to the application of EVs as biomarkers of both diseases. Key messages: Circulating EVs, specifically TF-positive EVs, in combination with plasmatic markers of hypercoagulable states, such as D-dimer, sP-selectin and antigen TF, are able to discriminate between cancer patients without thrombosis and patients with unprovoked VTD. Research fields could be opened. Future studies will assess if these biomarkers together serve as predicting thrombotic events in cancer populations.

Inhibition of the epigenetic suppressor EZH2 primes osteogenic differentiation mediated by BMP2.

Bone-stimulatory therapeutics include bone morphogenetic proteins (e.g. BMP2), parathyroid hormone, and antibody-based suppression of WNT antagonists. Inhibition of the epigenetic enzyme enhancer of zeste homolog 2 (EZH2) is both bone anabolic and osteoprotective. EZH2 inhibition stimulates key components of bone-stimulatory signaling pathways, including the BMP2 signaling cascade. Because of high costs and adverse effects associated with BMP2 use, here we investigated whether BMP2 dosing can be reduced by co-treatment with EZH2 inhibitors. Co-administration of BMP2 with the EZH2 inhibitor GSK126 enhanced differentiation of murine (MC3T3) osteoblasts, reflected by increased alkaline phosphatase activity, Alizarin Red staining, and expression of bone-related marker genes (e.g. Bglap and Phospho1). Strikingly, co-treatment with BMP2 (10 ng/ml) and GSK126 (5 µm) was synergistic and was as effective as 50 ng/ml BMP2 at inducing MC3T3 osteoblastogenesis. Similarly, the BMP2-GSK126 co-treatment stimulated osteogenic differentiation of human bone marrow-derived mesenchymal stem/stromal cells, reflected by induction of key osteogenic markers (e.g. Osterix/SP7 and IBSP). A combination of BMP2 (300 ng local) and GSK126 (5 µg local and 5 days of 50 mg/kg systemic) yielded more consistent bone healing than single treatments with either compound in a mouse calvarial critical-sized defect model according to results from µCT, histomorphometry, and surgical grading of qualitative X-rays. We conclude that EZH2 inhibition facilitates BMP2-mediated induction of osteogenic differentiation of progenitor cells and maturation of committed osteoblasts. We propose that epigenetic priming, coupled with bone anabolic agents, enhances osteogenesis and could be leveraged in therapeutic strategies to improve bone mass.

Genotoxicity in fishes environmentally exposed to As, Se, Hg, Pb, Cr and toxaphene in the lower Colorado River basin, at Mexicali valley, Baja California, México.

The environmental exposure to As, Se, Hg, Pb, Cr and toxaphene was assessed for 11 freshwater fish species in irrigation channels, agricultural return flow drains, a drain collecting lagoon and sections of the Colorado River at the Mexicali valley in Baja California, México, during August 2015-April 2016. Arsenic (2.90 ng ml-1) and Se (1.41 ng ml-1) in water had the highest concentrations in the return flow drains (Hardy River and Xochimilco Lagoon, respectively). However, fish axial muscle tissue had the highest concentration of Se (8.3 µg g-1) and Hg (0.36 µg g-1) in Colorado River fresh water, while As (1.7 µg g-1) in Hardy River fish was highest. Selenium concentrations in all fishes and toxaphene in Cyprinus carpio and Ameiurus natalis are above the safe levels for human consumption (0.3 µg g-1 and 180 ng g-1 respectively). Toxaphene was detected in the fish axial tissue, having the highest concentrations in Poecilia latipinna (690 ng g-1) in the Colorado River. The low proportion of the 8-Cl toxaphene congeners in fish suggests degradation of this pollutant. Tilapia. sp. cf. zillii had the most genotoxic damage with 7.4 micronucleated erythrocytes per 10,000 erythrocytes in Xochimilco Lagoon and 2 in Hardy River. The genotoxicity in all the fish species studied was significantly correlated to the concentrations of As and Se in water.

[Cystatin C: Biomarker of cardiovascular risk in HIV]. / Cistatina C: Biomarcador de riesgo cardiovascular en VIH.

INTRODUCTION: Patients infected with the human immunodeficiency virus (HIV) have a higher cardiovascular risk (CVR). The development of cardiovascular disease (CVD) in this population involves traditional CVR factors and factors related to the infection itself, such as chronic inflammatory status, immune dysfunction, as well as the antiretroviral therapy received. Cystatin C (CC) has shown to be useful in assessing the presence of CVR factors and CVD established in the general population, the elderly population, and patients with chronic kidney disease. An analysis was performed on this association in an HIV positive population (HIV+). MATERIAL AND METHODS: Analytical, observational, cross-sectional study was conducted, and included collecting information about CVR factors and CVD in HIV+, as well as measuring CC. The patients were divided into 2 groups: Group1=high CC (≥0.95mg/L) and Group2=normal CC (<0.95mg/L). RESULTS: A total of 95 patients were included. Group1=27 (28.4%) and Group2=68 (71.5%). A value of CC≥0.95mg/L was related to the presence of CVD (P=.01). It was also related with and an increase in waist circumference (P=.05), neck circumference (P=.04), systolic blood pressure (P=.04), diastolic blood pressure (P=.01), Framingham score (P=.03), and Framingham score adapted for HIV (P=.01). After performing multivariate analysis with incorporation of variables associated with CVD in the bivariate analysis, only CC≥0.95mg/L continued to be related to CVD. CONCLUSION: CC≥0.95mg/L was independently associated with CVD. This cut-off point was also linked to higher levels of blood pressure, and higher CVR at 10 years using the Framingham Score and Framingham Score adapted for HIV population.

Hypothermia and nutrient deprivation alter viability of human adipose-derived mesenchymal stem cells.

PURPOSE: Adipose-derived mesenchymal stem cells (MSCs) are attractive biological agents in regenerative medicine. To optimize cell therapies, it is necessary to determine the most effective delivery method for MSCs. Therefore, we evaluated the biological properties of MSCs after exposure to various temperatures to define optimal storage conditions prior to therapeutic delivery of MSCs. DESIGN: Prospective observational study. METHODS AND MATERIALS: Adherent and non-adherent MSCs were incubated at multiple temperatures (i.e., 4, 23 and 37 °C) in Lactated Ringers (LR) solution lacking essential cell growth ingredients, or in culture media which is optimized for cell growth. Cells were assessed either after the temperature changes (4 h) or after recovery (24 h). Metabolic activity of MSCs, cell number and expression of representative mRNA biomarkers were evaluated to assess the biological effects of temperature. We monitored changes in mRNAs expression related to cytoprotective- or stress-related responses (e.g., FOS, JUN, ATF1, ATF4, EGR1, EGR2, MYC), proliferation (e.g., HIST2H4, CCNB2), and extracellular matrix production (ECM; e.g., COL3A1, COL1A1) by quantitative real time reverse-transcriptase polymerase chain reaction (RT-qPCR) analysis. RESULTS: Our study demonstrates that storing MSCs in Lactated Ringers (LR) solution for 4 h decreases cell number and metabolic activity. The number of viable MSCs decreased significantly when cultured at physiological temperature (37 °C) and severe hypothermia (4 °C), while cells grown at ambient temperature (23 °C) exhibited the least detrimental effects. There were no appreciable biological differences in mRNA markers for proliferation or ECM deposition at any of the temperatures. However, biomarkers related to cytoprotective- or stress-responses were selectively elevated depending on temperature or media type (i.e., LR versus standard media). CONCLUSION: The biological impact of nutrient-free media and temperature changes after 4 h exposure persists after a 24 h recovery period. Hence, storage temperature and media conditions should be optimized to improve effective dosing of MSCs.

The epigenetic reader Brd4 is required for osteoblast differentiation.

Transcription networks and epigenetic mechanisms including DNA methylation, histone modifications, and noncoding RNAs control lineage commitment of multipotent mesenchymal progenitor cells. Proteins that read, write, and erase histone tail modifications curate and interpret the highly intricate histone code. Epigenetic reader proteins that recognize and bind histone marks provide a crucial link between histone modifications and their downstream biological effects. Here, we investigate the role of bromodomain-containing (BRD) proteins, which recognize acetylated histones, during osteogenic differentiation. Using RNA-sequencing (RNA-seq) analysis, we screened for BRD proteins (n = 40) that are robustly expressed in MC3T3 osteoblasts. We focused functional follow-up studies on Brd2 and Brd4 which are highly expressed in MC3T3 preosteoblasts and represent "bromodomain and extra terminal domain" (BET) proteins that are sensitive to pharmacological agents (BET inhibitors). We show that small interfering RNA depletion of Brd4 has stronger inhibitory effects on osteoblast differentiation than Brd2 loss as measured by osteoblast-related gene expression, extracellular matrix deposition, and alkaline phosphatase activity. Similar effects on osteoblast differentiation are seen with the BET inhibitor +JQ1, and this effect is reversible upon its removal indicating that this small molecule has no lasting effects on the differentiation capacity of MC3T3 cells. Mechanistically, we find that Brd4 binds at known Runx2 binding sites in promoters of bone-related genes. Collectively, these findings suggest that Brd4 is recruited to osteoblast-specific genes and may cooperate with bone-related transcription factors to promote osteoblast lineage commitment and maturation.

Análisis de factores psicológicos en pacientes con asma grave (estudio PSICOASMA) / Analysis of psychological factors in patients with severe asthma (PSICOASMA study)

OBJETIVOS: estudiar factores psicológicos en pacientes con asma grave y evaluar su relación con el control del asma. METODOLOGÍA: estudio prospectivo con inclusión de pacientes, diagnosticados de asma bronquial grave, revisados en consultas monográficas, participantes en el estudio de intervención psicológica (PSICOASMA). Se analizaron datos clínicos, cuestionario de control del asma (ACT), espirometría, inflamación (FENO). Se estudiaron alteraciones psicológicas con el cuestionario de ansiedad rasgo y estado (STAI-R y E), el inventario de depresión de BECK (BDI), cuestionario de Nijmegen. Se consideró alterado una puntuación en percentil >50 en el cuestionario STAI, Síndrome Hiperventilación si presentaban Nijmegen>23. RESULTADOS: Fueron incluidos 103 asmáticos graves, de edad 51,6 (13,7), mayoritariamente mujeres (79,6%). El 78,6% presentaban mal control del asma (ACT<20).Presentaban alterados test STAI-E, STAI-R el 50,5% y 68%, respectivamente y Nijmegen 47,5%. Respecto al nivel de depresión, según BDI, presentaron alteraciones leves, moderadas e intensas en el 31%, 25,2% y 11,6%, respectivamente. Se encontraron diferencias significativas en el control del asma medido por ACT entre los pacientes que presentaban o no alterados los test de ansiedad; STAI-E (p <0,05), STAI- R (p <0,01); depresión (p <0,01) o Nijmegen (p <0,01), estando peor controlados los que tenían alteración en los test psicológicos. No se encontraron diferencias en parámetros funcionales ni cifras de FENO. CONCLUSIONES: encontramos niveles elevados de ansiedad y depresión en pacientes con asma grave. Presentaban disnea funcional casi la mitad de los pacientes. Estas alteraciones psicológicas están relacionadas con el mal control de la enfermedad. Es importante y necesaria la evaluación psicológica en pacientes con asma grave

OBJECTIVES: to study the psychological factors in patients with severe asthma and evaluate their relationship with asthma control. METHODS: a prospective study including patients diagnosed with severe bronchial asthma who were examined in specialized clinics and who participated in the PSICOASMA psychological intervention study. The clinical data, asthma control test (ACT), spirometry and inflammation (FeNO) were analyzed. Psychological changes were studied using the State-Trait Anxiety Inventory (STAI-r and STAI-e), the Beck Depression Inventory (BDI) and the Nijmegen Questionnaire. A percentile score of >50 on the STAI was considered altered, and hyperventilation syndrome was determined for Nijmegen >23. RESULTS: 103 primarily female (79.6%) severely asthmatic patients were included with an average age of 51.6 (13.7). 78.6% showed poor asthma control (ACT <20). 50.5% and 68% showed altered STAI-e and STAI-r results, respectively, and 47.5% showed altered Nijmegen results. With regard to the degree of depression, according to the BDI, 31% of patients showed mild changes, 25.2% moderate changes and 11.6% intense changes. Significant differences were found in asthma control measured by ACT between patients with or without altered results in the anxiety test; STAI-e (p <0.05), STAI-r (p <0.01); depression questionnaire (p <0.01) and Nijmegen Questionnaire (p <0.01), with those showing alteration in the psychological tests being more poorly controlled. No differences were found in functional parameters or FeNO figures. CONCLUSIONS: We found high levels of anxiety and depression in patients with severe asthma. Nearly half of patients showed functional dyspnea. These psychological changes are related to the poor control of the disease. Psychological evaluation is important and necessary in patients with severe asthma

Causas de fallecimiento tras una media de 14 años de seguimiento en 1110 pacientes con asma (cohorte estudio perinasma) / Causes of death after an average follow-up of 14 years in 1110 patients with asthma (perinasma cohort study)

OBJETIVO: estudiar las causas de muerte en pacientes diagnosticados con asma en consultas especializadas tras una media de 14 años de seguimiento. METODOLOGÍA: se realizó un seguimiento (1998 - 2016) de 1.110 pacientes diagnosticados por primera vez de asma en consultas monográficas de asma bronquial. Se contactó telefónicamente con los pacientes. En caso de fallecimiento, buscamos la causa de muerte, según los informes de éxitus. Evaluamos estas causas en el plazo temporal referido. Los datos se expresan en % y se realizó análisis estadístico, estudiando las diferencias cualitativas con prueba de chi cuadrado, así como de las medias (DE), mediante t-student. RESULTADOS: del total estudiado, 690 pacientes (62,2%) eran mujeres (edad media 37,8 (17,6)) años y 420 (37,8%) hombres (edad media 29,5 (13,9) años, (p <0,01). Un 64,6% eran atópicos, 348 (31,4%) diagnosticados sólo de asma y 762 (68,6%) de asma con rinitis. En el periodo evaluado habían fallecido un total de 54 personas (4,9%), 46 mujeres (7,14%) y 8 hombres (1,9%). Las causas de fallecimiento fueron: 22 casos por neoplasias (40,7%), 9 casos de enfermedades del sistema circulatorio (16,6%), 2 casos por enfermedad neurodegenerativa (3,7%), 1 caso por infección (1,8%), 1 caso por asma e insuficiencia respiratoria (1,8%), 1 otras causas (1,8%) y 17 no conocido (31,4%). CONCLUSIONES: la causa más frecuente de fallecimiento de nuestros pacientes asmáticos fueron las neoplasias. El número de fallecimientos por enfermedades del sistema circulatorio fue bajo, inferior a la población general. Sólo hubo un caso recogido de fallecimiento por crisis de asma

OBJECTIVE: to study the cause of death in patients diagnosed with asthma in specialized outpatient clinics after an average follow-up of 14 years. METHODS: 1110 patients diagnosed with asthma for the first time in clinics specializing in bronchial asthma were monitored from 1998 to 2016. Patients were contacted by phone. In case of death, cause of death was determined according to the death certificate. These causes were evaluated during the abovementioned time period. The data is expressed as % and a statistical analysis was performed, studying the qualitative differences using the chi-square test as well as standard deviation using the student's t-test. RESULTS: of all patients studied, 690 (62.2%) were women (average age: 37.8 [17.6]) and 420 (37.8%) were men (average age: 29.5 [13.9]) (p <0.01). 64.6% were atopic, 348 (31.4%) were diagnosed only with asthma and 762 (68.6%) with asthma and rhinitis. During the evaluation period, a total of 54 patients died (4.9%): 46 women (7.14%) and 8 men (1.9%). The causes of death were: 22 cases due to neoplasm (40.7%), 9 cases due to circulatory system disease (16.6%), 2 cases due to neurodegenerative disease (3.7%), 1 case due to infection (1.8%), 1 case due to asthma and respiratory distress (1.8%), 1 case due to other causes (1.8%) and the cause of death was unknown in 17 patients (31.4%). CONCLUSIONS: the most frequent cause of death in our asthmatic patients was neoplasm. The number of deaths due to circulatory system disease was low, below the general population. Only one case of death due to asthma attack was recorded