Evaluation of single-cell RNAseq labelling algorithms using cancer datasets.

Single-cell RNA sequencing (scRNA-seq) clustering and labelling methods are used to determine precise cellular composition of tissue samples. Automated labelling methods rely on either unsupervised, cluster-based approaches or supervised, cell-based approaches to identify cell types. The high complexity of cancer poses a unique challenge, as tumor microenvironments are often composed of diverse cell subpopulations with unique functional effects that may lead to disease progression, metastasis and treatment resistance. Here, we assess 17 cell-based and 9 cluster-based scRNA-seq labelling algorithms using 8 cancer datasets, providing a comprehensive large-scale assessment of such methods in a cancer-specific context. Using several performance metrics, we show that cell-based methods generally achieved higher performance and were faster compared to cluster-based methods. Cluster-based methods more successfully labelled non-malignant cell types, likely because of a lack of gene signatures for relevant malignant cell subpopulations. Larger cell numbers present in some cell types in training data positively impacted prediction scores for cell-based methods. Finally, we examined which methods performed favorably when trained and tested on separate patient cohorts in scenarios similar to clinical applications, and which were able to accurately label particularly small or under-represented cell populations in the given datasets. We conclude that scPred and SVM show the best overall performances with cancer-specific data and provide further suggestions for algorithm selection. Our analysis pipeline for assessing the performance of cell type labelling algorithms is available in https://github.com/shooshtarilab/scRNAseq-Automated-Cell-Type-Labelling.

Modified Endaural Approach With a Peripheral Dissection of the Superficial Musculoaponeurotic System to Access the Temporomandibular Joint to Prevent Facial Nerve Injury.

The purpose of this study was to assess the effects of protecting the facial nerve with a modified endaural approach with a peripheral dissection of the superficial musculoaponeurotic system to access the temporomandibular joint which allows an excellent operative field of visualization, multiple surgical procedures of the temporomandibular joint, and general care which improves the immediate postoperative period, making this a less morbid surgery. This study included 33 patients (39 sides) who underwent surgical treatment for disorders of the temporomandibular joint from the years 2021 to 2023 at the maxillofacial department of the clinic "Colsanitas" located in Bogota, Colombia. Therapeutic results were evaluated by postoperative facial nerve injury, with the House-Brackman scale; every patient was examined for adequate facial musculature function immediately after surgery. Notably, zero patients presented facial nerve injury. These results imply that the modified endaural approach with a peripheral dissection of the superficial musculoaponeurotic system to access the temporomandibular joint reduces the incidence of facial nerve injuries, improves operative site exposure, and lowers the frequency of complications.

Absent right common carotid artery with associated anterior communicating aneurysm: embryology and imaging correlation.

PURPOSE: To report an unusual absent right common carotid artery with embryological and clinical emphasis. METHODS: A 63-year-old female with a multilobulated anterior communicating artery saccular aneurysm was referred to our center for definite treatment. An analysis of her carotid system on CTA and DSA with 3D modeling was performed to assess the embryology of an absent right common carotid and its association with aneurysm development. RESULTS: Cerebral angiogram demonstrated an absent right common carotid artery and separate origin of the internal and external right carotid arteries arising from the brachiocephalic trunk. CONCLUSION: Absence of the common carotid artery is an uncommon anatomical variant, usually asymptomatic and commonly associated with other vascular abnormalities. It is important to be familiar with the association between this anatomical variant given its hemodynamic stress, high risk of stroke, and aneurysm formation.

A Pleiotropic Flowering Time QTL Exhibits Gene-by-Environment Interaction for Fitness in a Perennial Grass.

Appropriate flowering time is a crucial adaptation impacting fitness in natural plant populations. Although the genetic basis of flowering variation has been extensively studied, its mechanisms in nonmodel organisms and its adaptive value in the field are still poorly understood. Here, we report new insights into the genetic basis of flowering time and its effect on fitness in Panicum hallii, a native perennial grass. Genetic mapping in populations derived from inland and coastal ecotypes identified flowering time quantitative trait loci (QTL) and many exhibited extensive QTL-by-environment interactions. Patterns of segregation within recombinant hybrids provide strong support for directional selection driving ecotypic divergence in flowering time. A major QTL on chromosome 5 (q-FT5) was detected in all experiments. Fine-mapping and expression studies identified a gene with orthology to a rice FLOWERING LOCUS T-like 9 (PhFTL9) as the candidate underlying q-FT5. We used a reciprocal transplant experiment to test for local adaptation and the specific impact of q-FT5 on performance. We did not observe local adaptation in terms of fitness tradeoffs when contrasting ecotypes in home versus away habitats. However, we observed that the coastal allele of q-FT5 conferred a fitness advantage only in its local habitat but not at the inland site. Sequence analyses identified an excess of low-frequency polymorphisms at the PhFTL9 promoter in the inland lineage, suggesting a role for either selection or population expansion on promoter evolution. Together, our findings demonstrate the genetic basis of flowering variation in a perennial grass and provide evidence for conditional neutrality underlying flowering time divergence.

The state of art of awake craniotomy in Latin American countries: a scoping review.

BACKGROUND AND OBJECTIVE: Awake craniotomy (AC) is a valuable technique for surgical interventions in eloquent areas, but its adoption in low- and middle-income countries faces challenges like limited infrastructure, trained personnel shortage, and inadequate funding. This scoping review explores AC techniques in Latin American countries, focusing on patient characteristics, tumor location, symptomatology, and outcomes. METHODS: A scoping review followed PRISMA guidelines, searching five databases in English, Spanish, and Portuguese. We included 28 studies with 258 patients (mean age: 43, range: 11-92). Patterns in AC use in Latin America were analyzed. RESULTS: Most studies were from Brazil and Mexico (53.6%) and public institutions (70%). Low-grade gliomas were the most common lesions (55%), most of them located in the left hemisphere (52.3%) and frontal lobe (52.3%). Gross-total resection was achieved in 34.3% of cases. 62.9% used an Asleep-Awake-Asleep protocol, and 14.8% used Awake-Awake-Awake. The main complication was seizures (14.6%). Mean post-surgery discharge time was 68 h. Challenges included limited training, infrastructure, and instrumentation availability. Strategies discussed involve training in specialized centers, seeking sponsorships, applying for awards, and multidisciplinary collaborations with neuropsychology. CONCLUSION: Improved accessibility to resources, infrastructure, and adequate instrumentation is crucial for wider AC availability in Latin America. Despite disparities, AC implementation with proper training and teamwork yields favorable outcomes in resource-limited centers. Efforts should focus on addressing challenges and promoting equitable access to this valuable surgical technique in the region.

Bridging Validity Frameworks in Assessment: Beyond Traditional Approaches in Health Professions Education.

Construct: High-stakes assessments measure several constructs, such as knowledge, competencies, and skills. In this case, validity evidence for test scores' uses and interpretations is of utmost importance, because of the consequences for everyone involved in their development and implementation. Background: Educational assessment requires an appropriate understanding and use of validity frameworks; however, health professions educators still struggle with the conceptual challenges of validity, and frequently validity analyses have a narrow focus. Important obstacles are the plurality of validity frameworks and the difficulty of grounding these abstract concepts in practice. Approach: We reviewed the validity frameworks literature to identify the main elements of frequently used models (Messick and Kane's) and proposed linking frameworks including Russell's recent overarching proposal. Examples are provided with commonly used assessment instruments in health professions education. Findings: Several elements in these frameworks can be integrated into a common approach, matching and aligning Messick's sources of validity with Kane's four inference types. Conclusions: This proposal to contribute evidence for assessment inferences may provide guidance to understanding the use of validity evidence in applied settings. The evolving field of validity research provides opportunities for its integration and practical use in health professions education.

Acellular Dermal Matrix, Osseoplastic Flap, and Transnasal Endoscopic Approach for Frontal Mucocele.

PURPOUSE: The primary objective of this study is to suggest the use of acellular dermal matrix through an osteoplastic flap and transnasal endoscopic treatment for the management of frontal mucoceles. The secondary objective is to propose the characteristics to choose this approach. METHODS: A retrospective cohort study was carried out on patients with frontal mucoceles of different etiologies where an osteoplastic flap was made to address the sinus and subsequently obliterated with acellular dermal matrix. RESULTS: A cohort of 11 patients were included in the study, 6 (67%) were female and 5 (56%) were male, with a mean age of 56 years (range 35-71). The majority of patients (73%) with a history of trauma and all the patients were treated with frontal osteoplastic flap and obliteration with acellular dermal matrix. No evidence of recurrence in a follow-up period with a mean of 18 months and a low rate of complications. CONCLUSIONS: The frontal osteoplastic flap and obliteration with acellular dermal matrix is a simple and safe technique to perform with low morbidity. Also, an orbital reconstruction can be performed simultaneously.

Antibiotic prescription errors: the relationship with clinical competence in junior medical residents.

BACKGROUND: A large portion of prescribing errors can be attributed to deficiencies in medication knowledge. These errors are preventable and most often occur at the time of prescription. Antimicrobials are the drug class most common incorrectly prescribed. OBJECTIVE: To characterize the relationship between clinical competence and antibiotic prescription errors. We also investigated the frequency and severity of antibiotic prescription errors to identify items and attributes of clinical competence which are correlated with the antibiotic prescription error ratio. METHOD: A cross-sectional study was applied to assess clinical competence of junior medical residents in two reference academic hospitals and a regional hospital in Mexico City. It was conducted during February 2019. We used an infectious disease Objective Structured Clinical Examination (OSCE) to assess clinical competence and a measure of frequency, and severity of antibiotic prescription errors. RESULTS: The number of eligible participants was ~ 255 (hospital meeting attendance), and the number of residents in this study were 51 (~ 20%), 31 were female (60.8%). The mean OSCE score was 0.692 ± 0.073. The inter-item (Cronbach's alpha = 0.927) and inter-station internal consistency was adequate (Cronbach's alpha = 0.774). The G coefficient in generalizability theory analysis was 0.84. The antibiotic prescription error ratio was 45.1% ± 7%. The most frequent category of severity of antibiotic prescription errors was category E (errors that may contribute to or result in temporary harm to the patient and require intervention), 235 (65.2%). We observed a negative and significant correlation between clinical competence and antibiotic prescription errors (r = -0.33, p < 0.05, CI95% -0.57 to -0.07), which remained significant after controlling for the effect of gender and time since graduation from medical school (r = -0.39, p < 0.01, CI95% -0.625 to -0.118). Using exploratory factor analysis we identified two factors, which explained 69% of the variance in clinical competence, factor 1 evaluated socio-clinical skills and factor 2 evaluated diagnostic-therapeutic skills. Factor 2 was correlated with antibiotic prescription error ratio (r = -0.536, p < 0.001). CONCLUSIONS: We observed a negative correlation between clinical competence and antibiotic prescription error ratio in graduated physicians who have been accepted in a medical specialty. The therapeutic plan, which is a component of the clinical competence score, and the prescription skills had a negative correlation with antibiotic prescription errors. The most frequent errors in antibiotic prescriptions would require a second intervention.

Does molar-incisor hypomineralization (MIH) affect only permanent first molars and incisors? New observations on permanent second molars.

BACKGROUND: The second permanent molar plays an important role when considering extraction of first permanent molars severely affected by molar-incisor hypomineralization (MIH). AIM: To assess the association between MIH and enamel hypomineralization of the second permanent molars in terms of presence and severity. DESIGN: In this retrospective cross-sectional study, permanent teeth were evaluated in 453 patients between the ages of 13 and 16 at a dental clinic, using intraoral photographs. A calibrated examiner classified enamel hypomineralization using the MIH index and dental caries experience using the DMFT and DMFS indices. Statistical analyses were performed using a linear generalized model and ordinal logistic regression. RESULTS: The most commonly affected teeth by enamel hypomineralization were permanent molars, whereas incisors, premolars, and canines were less affected. The presence of severe defects in the first permanent molars was associated with mild defects in the second permanent molars (OR = 4.01; 95% CI: 2.50-7.77). Enamel hypomineralization was associated with increased caries experience (PR = 5.54; 95% CI: 3.81-9.06). CONCLUSION: Enamel hypomineralization mainly affects the first and second permanent molars. Mild defects in the second permanent molars tend to be more frequent in patients with severe MIH in the first permanent molars. The presence of enamel hypomineralization was associated with higher dental caries experience.

Survival of stainless-steel crowns and composite resin restorations in molars affected by molar-incisor hypomineralization (MIH).

BACKGROUND: Currently, there is no consensus on the superiority of any material for the restorative treatment of molars affected by molar-incisor hypomineralization (MIH). AIM: To evaluate the survival of restorations with stainless-steel crown (SSC) or composite resin (CR) in first permanent molars affected by MIH for 24 months. DESIGN: In this retrospective cohort study, 61 CR and 54 SSC restorations placed on molars affected by MIH of patients, aged between 7 and 10, that were treated and overseen at a university dental clinic in the period of 2017-2020 were evaluated. The primary outcome was the failure-free survival time. Parametric survival models were used for data censored by interval, and the comparison between SSC and CR was performed using the hazard ratio function with a 95% confidence interval. RESULTS: The survival of SSC and CR restorations after 24 months was 94.4% and 49.2%, respectively. This difference was influenced by the presence of previous restoration (aHR = 3.4; 95% CI: 1.2-9.4) and cusp involvement (aHR = 4.0; 95% CI: 1.5-11.2). CONCLUSION: In molars with MIH and the need for restorative treatment, SSC had a significantly higher survival rate than CR over 24 months.

High jugular bulb with a diverticulum and vestibular aqueduct dehiscence: an anatomical variant to be aware in patients with hearing loss.

PURPOSE: To describe an anatomical variant that should be consider in patients with hearing loss. METHODS: An 8-year-old girl underwent to temporal bone computed tomography for the evaluation of bilateral conductive hearing loss and further assessment of possible enlarged vestibular aqueduct or high jugular bulb on brain magnetic resonance imaging (MRI). RESULTS: CT of temporal bone showed a cystic cavity with bony sclerotic margins extending from the right jugular foramen to the vestibular aqueduct. Bony dehiscence of the jugular foramen with the right carotid canal was also noted. On brain MRI, there was no evidence of enlargement of the endolymphatic duct and sac on T2 thin-section gradient echo sequence. Time of flight MR angiography did not show arterial flow in the cavity. Contrast enhanced MR venography confirmed the presence of a high right jugular bulb with a diverticulum extending into the vestibular aqueduct due to jugular bulb-vestibular aqueduct dehiscence. CONCLUSION: Knowledge of high jugular bulb-vestibular aqueduct dehiscence is important in the assessment of patients with otologic symptoms such as vertigo, tinnitus and hearing loss.

Evaluation of the usefulness of vitamin D as a predictor of mortality in patients with COVID-19.

INTRODUCTION: One of the functions of vitamin D is to regulate respiratory epithelium inflammatory response; therefore, deficiency of this vitamin in the context of COVID-19 could constitute a predictive biomarker of the disease outcome. OBJECTIVE: To evaluate the usefulness of vitamin D for predicting mortality in patients with COVID-19. METHODS: Observational, retrospective study in which 154 patients diagnosed with COVID-19 were included, out of whom 111 survived and 43 died. Vitamin D concentration was determined in all of them. RESULTS: A log-rank p-value < 0.032 was obtained for survival when vitamin D concentration was used as a categorical variable (≤ 20 ng/mL and > 20 ng/mL). On Cox proportional analysis, age and vitamin D concentration were shown to be risk factors associated with mortality in patients with COVID-19 (age: HR = 1.036, 95% CI = 1.016-1.058, p < 0.001; vitamin D: HR (≤ 20 ng/mL and > 20 ng/mL) = 0.478, 95% CI = 0.237-0.966, p < 0.040). CONCLUSION: Age and vitamin D concentration were predictive factors for mortality in COVID-19-infected patients.

INTRODUCCIÓN: Una de las funciones de la vitamina D es regular la respuesta inflamatoria del epitelio respiratorio; por ello, la deficiencia de esa vitamina en el contexto de COVID-19 podría constituir un biomarcador preditivo del desenlace de COVID-19. OBJETIVO: Evaluar la utilidad de la vitamina D para predecir la mortalidad en pacientes con COVID-19. MÉTODOS: Estudio observacional y retrospectivo en el que se incluyeron 154 pacientes con diagnóstico de COVID-19, de los cuales 111 sobrevivieron y 43 fallecieron. En todos se determinó la concentración de vitamina D. RESULTADOS: Se obtuvo un valor log-rank de p < 0.032 para la supervivencia al utilizar la concentración de vitamina D como variable categórica (≤ 20 ng/mL y > 20 ng/mL). Mediante análisis proporcional de Cox se encontró que la edad y concentración de vitamina D mostraron ser factores de riesgo asociados a la mortalidad en pacientes con COVID-19 (edad: HR = 1.036, IC 95 % = 1.016-1.058, p < 0.001; vitamina D: HR ≤ 20 ng/mL y > 20 ng/mL = 0.478, IC 95 % = 0.237-0.966, p < 0.040). CONCLUSIÓN: La edad y la concentración de vitamina D constituyeron factores predictivos de mortalidad en pacientes infectados por COVID-19.

Efficacy of osimertinib plus bevacizumab in glioblastoma patients with simultaneous EGFR amplification and EGFRvIII mutation.

BACKGROUND: Amplification of EGFR and its active mutant EGFRvIII are common in glioblastoma (GB). While EGFR and EGFRvIII play critical roles in pathogenesis, targeted therapy with EGFR-tyrosine kinase inhibitors or antibodies has shown limited efficacy. To improve the likelihood of effectiveness, we targeted adult patients with recurrent GB enriched for simultaneous EGFR amplification and EGFRvIII mutation, with osimertinib/bevacizumab at doses described for non-small cell lung cancer. METHODS: We retrospectively explored whether previously described EGFRvIII mutation in association with EGFR gene amplification could predict response to osimertinib/bevacizumab combination in a subset of 15 patients treated at recurrence. The resistance pattern in a subgroup of subjects is described using a commercial next-generation sequencing panel in liquid biopsy. RESULTS: There were ten males (66.7%), and the median patient's age was 56 years (range 38-70 years). After their initial diagnosis, 12 patients underwent partial (26.7%) or total resection (53.3%). Subsequently, all cases received IMRT and concurrent and adjuvant temozolomide (TMZ; the median number of cycles 9, range 6-12). The median follow-up after recurrence was 17.1 months (95% CI 12.3-22.6). All patients received osimertinib/bevacizumab as a second-line intervention with a median progression-free survival (PFS) of 5.1 months (95% CI 2.8-7.3) and overall survival of 9.0 months (95% CI 3.9-14.0). The PFS6 was 46.7%, and the overall response rate was 13.3%. After exposure to the osimertinib/bevacizumab combination, the main secondary alterations were MET amplification, STAT3, IGF1R, PTEN, and PDGFR. CONCLUSIONS: While the osimertinib/bevacizumab combination was marginally effective in most GB patients with simultaneous EGFR amplification plus EGFRvIII mutation, a subgroup experienced a long-lasting meaningful benefit. The findings of this brief cohort justify the continuation of the research in a clinical trial. The pattern of resistance after exposure to osimertinib/bevacizumab includes known mechanisms in the regulation of EGFR, findings that contribute to the understanding and targeting in a stepwise rational this pathway.

The Woven EndoBridge device for ruptured intracranial aneurysms: international multicenter experience and updated meta-analysis.

PURPOSE: The Woven EndoBridge (WEB) can be used to treat wide-necked aneurysms without antiplatelet medications, suggesting it may have advantages in the setting of aneurysmal subarachnoid hemorrhage (aSAH). The goal was assessment of safety and efficacy of WEB in aSAH given the delayed nature of aneurysmal thrombosis. METHODS: An international retrospective analysis of patients with aSAH treated with WEB was conducted at 7 tertiary centers from 2016 to 2020. Outcomes included rates of rebleeding, retreatment, complications, and complete occlusion. Furthermore, a systematic review and meta-analysis was conducted from 2011 to 2020 assessing the same outcomes. All pooled event rates were calculated using a random effect model. RESULTS: Consecutive patients with aSAH harbored 25 aneurysms that were treated with 29 WEB devices. The mean age was 53 years, and 65% were female. Zero experienced rebleeding, 2 were retreated, 2 experienced complications, 16 were completely occluded at 3 months, and 21 were completed occluded at 9-12 months. Meta-analysis of 309 WEB treatments for aSAH from 7 case series revealed 2.5% (95% CI 1-5%) had rebleeding, 9% (95% CI 4-17%) were retreated, 17% (95% CI 10-30%) had complications, and 61% (95% CI 51-71%) were completely occluded at 3-6 months. CONCLUSION: WEB embolization in the setting of aSAH provides similar protection against rebleeding with comparable retreatment rates to traditional approaches. However, there is a higher rate of incomplete radiographic occlusion and operative complications compared to WEB embolization of unruptured aneurysms. Long-term prospective studies are needed to fully delineate the role of WEB embolization in aSAH.

Endovascular treatment of anterior cranial fossa dural arteriovenous fistula: a multicenter series.

PURPOSE: We report a multicenter experience using endovascular embolization as the first line approach for treatment of anterior cranial fossa (ACF) dural arteriovenous fistula (DAVF). METHODS: All patients with DAVFs located in the anterior cranial fossa who were treated with endovascular technique as a first line approach were included. Demographics, clinical presentation, angioarchitecture, strategy, complications, immediate angiographic, and follow-up results were included in the analysis. RESULTS: Twenty-three patients met the inclusion criteria (18 male and 5 female). Age ranged from 14 to 79 years (mean 53 years). Twelve patients presented with hemorrhage. Twenty-eight endovascular procedures were performed. The overall immediate angiographic cure rate after endovascular treatment was 82.6% (19/23 patients). The angiographic cure rate of the transvenous strategy was significantly superior to the transarterial strategy (p ≤ 0.001). There was 1 complication in 28 total procedures (3.6%). Angiographic follow-up was available in 21 out of the 23 patients with a mean of 25 months (range 2 to 108 months). In these 21 patients, the DAVF was completely cured in 20 (95%). At last follow-up, all patients had a modified Rankin scale (mRS) 0 to 2. CONCLUSION: Our experience suggests that endovascular treatment for ACF DAVFs has an acceptable safety profile with high rates of complete occlusion, particularly with transvenous approach. Whenever possible, transvenous approach should be preferred over transarterial approach as first line strategy.

Anesthetic protocol for right ventricular dysfunction management in heart transplantation: systematic review, development and validation.

BACKGROUND: Right Ventricular Dysfunction (RVD) is the most frequent intraoperative hemodynamic complication in Heart Transplantation (HTx). RVD occurs in 0.04-1.0% of cardiac surgeries with cardiotomy and in 20-50% of HTx, with mortality up to 75%. No consensus has been established for how anesthesiologists should manage RVD, with management methods many times remaining unvalidated. METHODS: We conducted a systematic review, following PRISMA guidelines, to create an anesthetic protocol to manage RVD in HTx, using databases that include PubMed and Embase, until September 2018 based on inclusion and exclusion criteria. The articles screening for the systematic review were done two independent reviewers, in case of discrepancy, we consulted a third independent reviewer. Based on the systematic review, the anesthetic protocol was developed. The instrument selected to perform the validation of the protocol was AGREE II, for this purpose expert anesthetists were recruited to do this process. The minimum arbitration score for domains validation cutoff of AGREE II is arbitered to 70%. This study was registered at PROSPERO (115600). RESULTS: In the systematic review, 152 articles were included. We present the protocol in a flowchart with six steps based on goal-directed therapy, invasive monitoring, and transesophageal echocardiogram. Six experts judged the protocol and validated it. CONCLUSION: The protocol has been validated by experts and new studies are needed to assess its applicability and potential benefits on major endpoints.

PSYCOGENIC POLYDIPSIA AND SEVERE HYPONATREMIA IN BIPOLAR AFFECTIVE DISORDER.

Psychogenic polydipsia, primary polydipsia or potomania is a disorder of multifactorial etiology which is associated with substantial morbidity and mortality. It occurs frequently in patients with psychiatric diseases, particularly those with schizophrenia, however, it is not exclusive, it has been reported in a lower proportion in patients with anxiety disorders and mood disorders. Although, is still poorly understood and therefore underdiagnosed condition.

Role of RUNX2 transcription factor in epithelial mesenchymal transition in non-small cell lung cancer lung cancer: Epigenetic control of the RUNX2 P1 promoter.

Lung cancer has a high mortality rate in men and women worldwide. Approximately 15% of diagnosed patients with this type of cancer do not exceed the 5-year survival rate. Unfortunately, diagnosis is established in advanced stages, where other tissues or organs can be affected. In recent years, lineage-specific transcription factors have been associated with a variety of cancers. One such transcription factor possibly regulating cancer is RUNX2, the master gene of early and late osteogenesis. In thyroid and prostate cancer, it has been reported that RUNX2 regulates expression of genes important in tumor cell migration and invasion. In this study, we report on RUNX2/ p57 overexpression in 16 patients with primary non-small cell lung cancer and/or metastatic lung cancer associated with H3K27Ac at P1 gene promoter region. In some patients, H3K4Me3 enrichment was also detected, in addition to WDR5, MLL2, MLL4, and UTX enzyme recruitment, members of the COMPASS-LIKE complex. Moreover, transforming growth factor-ß induced RUNX2/ p57 overexpression and specific RUNX2 knockdown supported a role for RUNX2 in epithelial mesenchymal transition, which was demonstrated through loss of function assays in adenocarcinoma A549 lung cancer cell line. Furthermore, RUNX2 increased expression of epithelial mesenchymal transition genes VIMENTIN, TWIST1, and SNAIL1, which reflected increased migratory capacity in lung adenocarcinoma cells.

Differential placental ceramide levels during gestational diabetes mellitus (GDM).

BACKGROUND: Gestational diabetes mellitus (GDM) is associated with important factors that influence fetal development. Sphingolipids are known to be associated with the development of diabetes. Our objective was to examine ceramide, a key sphingolipid, hyperosmolarity, and apoptosis in placentas from GDM patients treated with insulin or diet. METHODS: Ceramide levels were assessed in placental tissues using immunohistochemistry. Immunoblot was performed to quantify serine palmitoyltransferase (SPT), the rate-limiting enzyme in ceramide biosynthesis, NFAT5, SMIT, AR, caspase 3 and the X-linked inhibitor of apoptosis. Trophoblast cells were treated with insulin or ceramide and assessments for mitochondrial respiration, caspase 3 and XIAP were also performed. RESULTS: Immunohistochemistry showed increased ceramides in the placental villous trophoblasts of the insulin-treated GDM patients. Nuclear SPT was upregulated only in the insulin-treated GDM placenta when compared to controls. Nuclear NFAT5 was also increased in the GDM placenta. Active caspase 3 was elevated in placentas from both insulin- and diet-treated GDM patients. Mitochondrial respiration was decreased in trophoblasts treated with ceramide. Active caspase was not changed while XIAP protein was increased in trophoblasts treated with ceramide. CONCLUSIONS: Our findings confirm the presence of ceramide in the human placenta of control and GDM patients. Furthermore, we conclude that ceramide is increased in the placental trophoblast during insulin treatment and that its upregulation correlates with elevated NFAT5, SMIT, increased apoptosis and decreased trophoblast mitochondrial respiration.