PKD2 regulates autophagy and forms a protein complex with BECN1 at the primary cilium of hypothalamic neuronal cells.

The primary cilium, hereafter cilium, is an antenna-like organelle that modulates intracellular responses, including autophagy, a lysosomal degradation process essential for cell homeostasis. Dysfunction of the cilium is associated with impairment of autophagy and diseases known as "ciliopathies". The discovery of autophagy-related proteins at the base of the cilium suggests its potential role in coordinating autophagy initiation in response to physiopathological stimuli. One of these proteins, beclin-1 (BECN1), it which is necessary for autophagosome biogenesis. Additionally, polycystin-2 (PKD2), a calcium channel enriched at the cilium, is required and sufficient to induce autophagy in renal and cancer cells. We previously demonstrated that PKD2 and BECN1 form a protein complex at the endoplasmic reticulum in non-ciliated cells, where it initiates autophagy, but whether this protein complex is present at the cilium remains unknown. Anorexigenic pro-opiomelanocortin (POMC) neurons are ciliated cells that require autophagy to maintain intracellular homeostasis. POMC neurons are sensitive to metabolic changes, modulating signaling pathways crucial for controlling food intake. Exposure to the saturated fatty acid palmitic acid (PA) reduces ciliogenesis and inhibits autophagy in these cells. Here, we show that PKD2 and BECN1 form a protein complex in N43/5 cells, an in vitro model of POMC neurons, and that both PKD2 and BECN1 locate at the cilium. In addition, our data show that the cilium is required for PKD2-BECN1 protein complex formation and that PA disrupts the PKD2-BECN1 complex, suppressing autophagy. Our findings provide new insights into the mechanisms by which the cilium controls autophagy in hypothalamic neuronal cells.

Pediatric anesthesiology in Brazil, Chile, and Mexico.

BACKGROUND: Latin America comprises an extensive and diverse territory composed of 33 countries in the Caribbean, Central, and South America where Romance languages-languages derived from Latin are predominantly spoken. Economic disparities exist, with inequitable access to pediatric surgical care. The Latin American Surgical Outcomes Study in Pediatrics (LASOS-Peds), a multi-national collaboration, will determine safety of pediatric anesthesia and perioperative care. OBJECTIVE: Below, we provide a descriptive initiative to share how pediatric anesthesia in Brazil, Chile, and Mexico operate. Theses descriptions do not represent all of Latin America. DESCRIPTIONS AND CONCLUSIONS: Brazil an upper middle-income country, population 203 million, has a public system insufficiently resourced and a private system, resulting in inequitable safety and accessibility. Surgical complications constitute the third leading cause of mortality. Anesthesiology residency is 3 years, with required rotations in pediatric anesthesia; five hospitals offer pediatric anesthesia fellowships. Anesthesiology is a physician-only practice. A Pediatric Anesthesia Committee within the Brazilian Society of Anesthesiology offers education through seasonal courses and workshops including pediatric advanced life support. Chile is a high-income country, population 19.5 million, the majority cared for in the public system, the remainder in university, private, or military systems. Government efforts have gradually corrected the long-standing anesthesiology shortage: twenty 3-year residency programs prepare graduates for routine pediatric cases. The Chilean Society of Anesthesiology runs a 1-month program for general anesthesiologists to enhance pediatric anesthesia skills. Pediatric anesthesia fellowship training occurs in Europe, USA, and Australia, or in two 2-year Chilean university programs. Public health policies have increased the medical and surgical pediatric specialists and general anesthesiologists, but not pediatric anesthesiologists, which creates safety concerns for neonates, infants, and medically complex. Chile needs more pediatric anesthesia fellowship programs. Mexico, an upper middle-income country, with a population of about 126 million, has a five-sector healthcare system: public, social security for union workers, state for public employees, armed forces for the military, and a private "self-pay." There are inequities in safety and accessibility for children. Pediatric Anesthesiology fellowship is 2 years, after 3 years residency. A shortage of pediatric anesthesiologists limits accessibility and safety for surgical care, driven by added training at low salary and hospital under appreciation. The Mexican Society of Pediatric Anesthesiology conducts refresher courses, workshops, and case conferences. Insufficient resources and culture limits research.

Measurable residual disease conversion rate with consolidation chemotherapy in acute myeloid leukemia.

The rate of MRD clearance in AML with standard consolidation chemotherapy is not well defined. A multi-institution retrospective analysis was performed on 107 consecutively treated AML patients in morphologic complete remission with detectable MRD post-induction therapy who received standard chemotherapy consolidation. In response to standard intermediate/high-dose cytarabine consolidation therapy, 26 of 60 patients (43.3%) with MRD threshold of detection of at least 0.1% converted to MRD-negative status (undetectable with assay used), and 6 of 47 patients (12.8%) with MRD threshold of detection > 0.1% converted to MRD-negative status. Multivariable logistic regression for patients with MRD threshold of detection of at least 0.1% showed that, when controlling for age, ELN risk category, dose of cytarabine, and use of a combination agent, treatment with 1 cycle of consolidation cytarabine versus ≥2 cycles decreased the odds of conversion of AML to MRD-negative (OR = 0.24, 95% CI 0.07-0.85, p = 0.03).

Progesterone and its metabolite allopregnanolone promote invasion of human glioblastoma cells through metalloproteinase­9 and cSrc kinase.

Glioblastomas are the most aggressive and common primary brain tumors in adults. Glioblastoma cells have a great capacity to migrate and invade the brain parenchyma, often reaching the contralateral hemisphere. Progesterone (P4) and its metabolite, allopregnanolone (3α-THP), promote the migration and invasion of human glioblastoma-derived cells. P4 induces migration in glioblastoma cells by the activation of the proto-oncogene tyrosine-protein kinase Src (cSrc) and focal adhesion kinase (Fak). In breast cancer cells, cSrc and Fak promote invasion by increasing the expression and activation of extracellular matrix metalloproteinases (MMPs). However, the mechanism of action by which P4 and 3a-THP promote invasion in glioblastoma cells remains unclear. The effects of P4 and 3α-THP on the protein expression levels of MMP-2 and -9 and the participation of cSrc in progestin effects in U251 and U87 human glioblastoma-derived cells were evaluated. It was determined by western blotting that the P4 increased the protein expression level of MMP-9 in U251 and U87 cells, and 3α-THP increased the protein expression level of MMP-9 in U87 cells. None of these progestins modified MMP-2 protein expression levels. The increase in MMP-9 expression was reduced when the intracellular progesterone receptor and cSrc expression were blocked with small interfering RNAs. Cell invasion induced by P4 and 3α-THP was also blocked by inhibiting cSrc activity with PP2 or by cSrc gene silencing. These results suggest that P4 and its metabolite 3α-THP induce the invasion of glioblastoma cells by increasing MMP-9 expression through the cSrc kinase family. The results of this study provide information of interest in the context of targeted therapies against molecular pathways involved in glioblastoma invasion.

Sex and Age Influence on Association of CYP450 Polymorphism with Midazolam Levels in Critically Ill Children.

Midazolam is a drug that is metabolized by cytochrome P450 (CYP450) enzymes, particularly CYP3A4 and CYP3A5. The present study aimed to determine the sex and age influence on association of CYP450 polymorphism with midazolam levels in critically ill children. Seventy-two DNA samples were genotyped by real-time PCR. Children ≤ five years of age who carry the rs776746 (T) allele in CYP3A5 gene were associated with lower plasma midazolam levels. The concentration median in patients was 0.0 ng/mL, while in patients with the normal (C) allele, it was 438.17 ng/mL (Q25 135.75-Q75 580.24), p = 0.005. The midazolam plasmatic concentration in female patients with the minor (T) allele was 0.0 ng/mL (Q250.00-Q75204.3), while in patients with the normal (C) allele median it was 459.0 ng/mL (Q25296.9-Q75789.7), p = 0.002. Analysis of the dominant model for the rs2740574 variant in CYP3A4 revealed a median of 0.38 L/kg (Q250.02-Q751.5) for the volume of distribution parameter in female patients with the normal T allele, while female patients with the minor C allele showed a median of 18.1 L/kg (Q257.5-Q7528.7) p = 0.02. Our results suggest an altered midazolam metabolism due to the presence the allelic rs2740574 variants of CYP3A4 and rs776746 of CYP3A5, and also the strong influence of age and sex.

Expression and estrogen regulation of G protein-coupled estrogen receptor in human glioblastoma cells.

Glioblastoma (GB) is the most frequent primary brain tumor with a very poor prognosis. Sex hormones are crucial players in the development of GBs. 17 ß-estradiol (E2) signaling is involved through its corresponding intracellular receptors [estrogen receptor α (ERα) and ß (ERß)] in GB cell proliferation and progression. E2 activates G-protein coupled estrogen receptor (GPER), leading to rapidly occurring effects, independently of gene transcription. GPER activation is involved in tumor progression in various cancer types. Currently, available data concerning the occurrence and role of GPER in GB are very limited. In the present study, it was observed that GPER was expressed in human brain tumor cell lines [U251 (astrocytoma-derived cell line), U87, LN229 and T98 (glioblastoma-derived cell line)]. Immunofluorescence assays revealed that GPER localizes in the plasma membrane, cytoplasm and nucleus. An in silico analysis identified two potential E2 response elements in the promoter region of the GPER gene. E2 increased GPER expression in the U251, U87 and LN229 cell lines. Molecular modeling data derived from in silico analysis predicted the three-dimensional conformation of GPER, and docking analysis identified potential binding sites of E2 and its specific agonist, G1. Taken together, these results indicate that GPER may be differentially expressed in human GB cell lines with E2 possibly upregulating GPER expression. The present study raises further questions about the implications of GPER-mediated E2 signaling in the biology of GBs.

Palmitic and Stearic Acids Inhibit Chaperone-Mediated Autophagy (CMA) in POMC-like Neurons In Vitro.

The intake of food with high levels of saturated fatty acids (SatFAs) is associated with the development of obesity and insulin resistance. SatFAs, such as palmitic (PA) and stearic (SA) acids, have been shown to accumulate in the hypothalamus, causing several pathological consequences. Autophagy is a lysosomal-degrading pathway that can be divided into macroautophagy, microautophagy, and chaperone-mediated autophagy (CMA). Previous studies showed that PA impairs macroautophagy function and insulin response in hypothalamic proopiomelanocortin (POMC) neurons. Here, we show in vitro that the exposure of POMC neurons to PA or SA also inhibits CMA, possibly by decreasing the total and lysosomal LAMP2A protein levels. Proteomics of lysosomes from PA- and SA-treated cells showed that the inhibition of CMA could impact vesicle formation and trafficking, mitochondrial components, and insulin response, among others. Finally, we show that CMA activity is important for regulating the insulin response in POMC hypothalamic neurons. These in vitro results demonstrate that CMA is inhibited by PA and SA in POMC-like neurons, giving an overview of the CMA-dependent cellular pathways that could be affected by such inhibition and opening a door for in vivo studies of CMA in the context of the hypothalamus and obesity.

EZH2 Mediates Proliferation, Migration, and Invasion Promoted by Estradiol in Human Glioblastoma Cells.

Glioblastomas (GBM) are the most frequent and aggressive brain tumors. 17ß-estradiol (E2) increases proliferation, migration, and invasion of human GBM cells; however underlying mechanisms are no fully understood. Zeste 2 Enhancer Homologous enzyme (EZH2) is a methyltransferase part of Polycomb 2 repressor complex (PRC2). In GBM, EZH2 is overexpressed and involved in the cell cycle, migration, and invasion processes. We studied the role of EZH2 in the pro-oncogenic actions of E2 in human GBM cells. EZH2 gene silencing and pharmacological inhibition of EZH2 blocked proliferation, migration, and invasion of GBM cells induced by E2. We identified in silico additional putative estrogen response elements (EREs) at the EZH2 promoter, but E2 did not modify EZH2 expression. In silico analysis also revealed that among human GBM samples, EZH2 expression was homogeneous; in contrast, the heterogeneous expression of estrogen receptors (ERs) allowed the classification of the samples into groups. Even in the GBM cluster with high expression of ERs and those of their target genes, the expression of PCR2 target genes did not change. Overall, our data suggest that in GBM cells, pro-oncogenic actions of E2 are mediated by EZH2, without changes in EZH2 expression and by mechanisms that appear to be unrelated to the transcriptional activity of ERs.

Hemophagocytic Lymphohistiocytosis in the Emergency Department: Recognizing and Evaluating a Hidden Threat.

BACKGROUND: Hemophagocytic lymphohistiocytosis (HLH) is a life-threatening hematologic disorder resulting from an ineffective and pathologic activation of the immune response system that may mimic common emergency department presentations, including sepsis, acute liver failure, disseminated intravascular coagulation, and flu-like illnesses such as coronavirus disease 2019 (COVID-19). OBJECTIVE: This narrative review provides a summary of the disease and recommendations for the recognition and diagnostic evaluation of HLH with a focus on the emergency clinician. DISCUSSION: Though the condition is rare, mortality rates are high, ranging from 20% to 80% and increasing with delays in treatment. Importantly, HLH has been recognized as a severe variation of the cytokine storm associated with COVID-19. Common features include a history of infection or malignancy, fever, splenomegaly or hepatomegaly, hyperferritinemia, cytopenias, coagulopathies, abnormal liver enzymes, and hypertriglyceridemia. Using specific features of the history, physical examination, laboratory studies, and tools such as the HScore, HLH-2004/2009, and hyperferritinemia thresholds, the emergency clinician can risk-stratify patients and admit for definitive testing. Once diagnosed, disease specific treatment can be initiated. CONCLUSION: This review describes the relevant pathophysiology, common presentation findings, and a framework for risk stratification in the emergency department.

Statistical approach to Trichinella infection in horses handled by rural slaughterhouses across five distinctive socioeconomic regions in Mexico

Abstract This work aimed to investigate the prevalence of Trichinella infection in horses (Equus ferus caballus) handled by rural slaughterhouses across five distinctive socioeconomic regions in Mexico. Serum samples were obtained by non-probabilistic convenience sampling in the Eastern, Southern Central and Western regions (100 samples of each). Additionally, muscle tissue samples were collected from the East (n = 45), Southeastern (n = 88), Southern Central (n = 39) and Southwestern (n = 11) regions. Antibodies were determined by Western blot and the muscle tissue was examined by artificial digestion. A global antibody prevalence of 2% was obtained. Regionally, a prevalence of 5% was observed in the East and 1% in the Southern Central region. No antibodies were detected in the West region and no larvae were found in the muscle tissue samples. These findings support the low presence of Trichinella in Mexican horses, which can positively impact the Mexican horse meat trade.

Resumen Se investigó la prevalencia de Trichinella en caballos (Equus ferus caballus) demataderos rurales en cinco regiones socioeconómicas de México. Los sueros se obtuvieron por Trichinella muestreo de conveniencia no probabilístico en las regiones Este, Centrosur y Oeste (100 muestras de cada una). Además se colectaron muestras de tejido muscular de animales de las regionesEste (n = 45), Centro Sur (n = 39), Sudeste (n = 88) y Sudoeste (n = 11). Los anticuerpos se determinaron por Western blot y el tejido muscular se examinó por digestión artificial. Se obtuvo una prevalencia global de anticuerpos del 2%. A nivel regional se observó una prevalencia del 5% enel Este y del 1% en el Centrosur. No se detectaron anticuerpos en el Oeste y no se encontraron larvas en el tejido muscular. Los datos respaldan la baja presencia de Trichinella en caballos mexicanos, lo que puede impactar positivamente en su comercio.

Statistical approach to Trichinella infection in horses handled by rural slaughterhouses across five distinctive socioeconomic regions in Mexico.

This work aimed to investigate the prevalence of Trichinella infection in horses (Equus ferus caballus) handled by rural slaughterhouses across five distinctive socioeconomic regions in Mexico. Serum samples were obtained by non-probabilistic convenience sampling in the Eastern, Southern Central and Western regions (100 samples of each). Additionally, muscle tissue samples were collected from the East (n=45), Southeastern (n=88), Southern Central (n=39) and Southwestern (n=11) regions. Antibodies were determined by Western blot and the muscle tissue was examined by artificial digestion. A global antibody prevalence of 2% was obtained. Regionally, a prevalence of 5% was observed in the East and 1% in the Southern Central region. No antibodies were detected in the West region and no larvae were found in the muscle tissue samples. These findings support the low presence of Trichinella in Mexican horses, which can positively impact the Mexican horse meat trade.

Recent Progress in Nanomaterials for Modern Concrete Infrastructure: Advantages and Challenges.

Modern concrete infrastructure requires structural components with higher mechanical strength and greater durability. A solution is the addition of nanomaterials to cement-based materials, which can enhance their mechanical properties. Some such nanomaterials include nano-silica (nano-SiO2), nano-alumina (nano-Al2O3), nano-ferric oxide (nano-Fe2O3), nano-titanium oxide (nano-TiO2), carbon nanotubes (CNTs), graphene and graphene oxide. These nanomaterials can be added to cement with other reinforcement materials such as steel fibers, glass, rice hull powder and fly ash. Optimal dosages of these materials can improve the compressive, tensile and flexural strength of cement-based materials, as well as their water absorption and workability. The use of these nanomaterials can enhance the performance and life cycle of concrete infrastructures. This review presents recent researches about the main effects on performance of cement-based composites caused by the incorporation of nanomaterials. The nanomaterials could decrease the cement porosity, generating a denser interfacial transition zone. In addition, nanomaterials reinforced cement can allow the construction of high-strength concrete structures with greater durability, which will decrease the maintenance requirements or early replacement. Also, the incorporation of nano-TiO2 and CNTs in cementitious matrices can provide concrete structures with self-cleaning and self-sensing abilities. These advantages could help in the photocatalytic decomposition of pollutants and structural health monitoring of the concrete structures. The nanomaterials have a great potential for applications in smart infrastructure based on high-strength concrete structures.

Low-temperature conditioning of "seed" cloves enhances the expression of phenolic metabolism related genes and anthocyanin content in 'Coreano' garlic (Allium sativum) during plant development.

Low-temperature conditioning of garlic "seed" cloves accelerated the development of the crop cycle, decreased plant growth, and increased the synthesis of phenolic compounds and anthocyanins in the outer scale leaves of the bulbs at harvest time, leading to 3-fold content increase compared with those conditioned at room temperature. Cold conditioning of "seed" cloves also altered the anthocyanin profile during bulb development and at harvest. Two new anthocyanins are reported for the first time in garlic. The high phenolics and anthocyanin contents in bulbs of plants generated from "seed" cloves conditioned at 5 °C for 5 weeks were preceded by overexpression of some putative genes of the phenolic metabolism [6-fold for phenylalanine ammonia lyase (PAL)] and anthocyanin synthesis [1-fold for UDP-sugar:flavonoid 3-O-glycosyltransferase (UFGT)] compared with those conditioned at room temperature.

Development and initial validation of the thyroid cancer module of the M. D. Anderson Symptom Inventory.

OBJECTIVE: The M. D. Anderson Symptom Inventory (MDASI) and its modules measure common symptoms related to cancer and its treatment. We report the development and initial validation of the MDASI-Thyroid Cancer module (MDASI-THY). METHODS: A list of thyroid-cancer-specific symptoms was generated through focus groups and interviews with thyroid cancer patients, clinicians and researchers. These MDASI-THY items were added to the original MDASI and administered to 60 patients with thyroid cancer. Symptom prevalence and severity were evaluated, along with the reliability and content, construct and known-group validity of the MDASI-THY. RESULTS: Cognitive debriefing performed on a subset of patients indicated that the MDASI-THY items were clear, concise, relevant and easy to understand. Fatigue, drowsiness, sleep disturbance, distress and difficulty remembering were the 5 most prevalent and severe symptoms. Twenty-eight percent of patients had moderate to severe fatigue (>or=5 on a 0-10 scale). Average severity was 1.28 and 1.29 for the symptom and interference subscales, respectively. MDASI-THY symptoms were severer for patients with poorer performance status. Cronbach alpha-values were 0.76, 0.85 and 0.92 for the thyroid-specific symptom items, core symptom subscale and interference subscale, respectively. CONCLUSIONS: This study demonstrates preliminary evidence for the validity and reliability of the MDASI-THY.