RESUMEN
PURPOSE: Various anatomical variations of the inferior alveolar canal increase the incidence of surgical complications; Therefore, this study aimed to evaluate the frequency and configuration of bifid and trifid mandibular canals using cone beam computed tomography (CBCT) in the Turkish subpopulation. METHODS: The inferior alveolar canal was evaluated on 1014 hemi-mandibles in the CBCT (I-CAT 3D Imaging System) images of 513 patients. The frequency and configuration of the bifid and trifid mandibular canal (MC) were examined. The relationship between bifid MC configuration and dental status and age groups was analyzed. The distance of the accessory canal to the buccal and lingual walls and the alveolar crest was measured. The diameter of the main canal and accessory canal was measured and its relationship with dental status and age groups was evaluated. RESULTS: Bifid MC was found in 266 hemi-mandibles (24.7%) and 212 (41.3%) of 513 patients. The most common type of bifid MC was the retromolar canal (87 sides), followed by the forward canal without confluence (41; 4%) and the dental canal (34; 3.4%). 10 of the dental canals were opening to the 1st molar, 14 of the 2nd molars, and 10 of the 3rd molars. The number of retromolar foramina was 1 on 56 sides, 2 on 15 sides, and 3 on 4 sides. Forward canal without confluence was more common in edentulous patients than in dentulous patients, while the dental canal was more common in dentulous patients. The main canal diameter was 3.53 ± 0.97 mm and the bifid MC diameter was 1.82 ± 0.70 mm. Distance of the bifid MC to the lingual wall was higher in the > 64 years group than in the 18-39 years group (p = 0.022). Distance of the bifid MC to the alveolar crest was lower in the > 64 years group compared to the 18-39 years group and 40-64 years group (p = 0.015). The main canal diameter was higher in the 40-64 years group than in the 18-39 years group (p = 0.012). CONCLUSION: Bifid MC has a high prevalence, occurring in almost one in two patients. Dental and retromolar types, which are close to the teeth, are more common, and this increases the possibility of complications. CBCT is the most accurate imaging technique used to detect and define these variations.
RESUMEN
BACKGROUND: We aimed to evaluate the validity and reliability of the Turkish version of the Patient-Reported Impact of Spasticity Measure (PRISM-TR). METHODS: Expert opinions and the forward-back translation method were used for linguistic validation. Cronbach α and test-retest analysis were performed for reliability analysis. Correlations between the PRISM-TR and the Modified Ashworth Scale (MAS), the Numerical Rating Scale (NRS), and the Expanded Disability Status Scale (EDSS) were assessed. RESULTS: A total of 206 individuals with multiple sclerosis (MS; 139 women; mean age, 47.7 ± 11.3 years; mean EDSS score, 5.2 ± 1.5) who had not had a relapse in the previous 3 months were included in the study. Test-retest correlation coefficients were high for all subdimensions of the scale (0.95-0.99). All Cronbach α values for the PRISM-TR subdimensions, except for Positive Impact, were likewise quite high (0.85-0.94). PRISM-TR subscale scores were correlated with MAS, NRS, and EDSS scores (P < .001). CONCLUSIONS: PRISM-TR is a valid and reliable scale for use with Turkish individuals with MS. It is simple to use in the clinic and can be helpful in detecting patients' spasticity early in the disease course.
RESUMEN
BACKGROUND: Falls in multiple sclerosis can result in numerous problems, including injuries and functional loss. Therefore, determining the factors contributing to falls in people with Multiple Sclerosis (PwMS) is crucial. This study aims to investigate the contributing factors to falls in multiple sclerosis using a machine learning approach. METHODS: This cross-sectional study was conducted with 253 PwMS admitted to the outpatient clinic of a university hospital between February and August 2023. A sociodemographic data collection form, Fall Efficacy Scale (FES-I), Berg Balance Scale (BBS), Fatigue Severity Scale (FSS), Expanded Disability Status Scale (EDSS), Multiple Sclerosis Impact Scale (MSIS-29), and Timed 25 Foot Walk Test (T25-FW) were used for data collection. Gradient-boosting algorithms were employed to predict the important variables for falls in PwMS. The XGBoost algorithm emerged as the best performed model in this study. RESULTS: Most of the participants (70.0%) were female, with a mean age of 40.44 ± 10.88 years. Among the participants, 40.7% reported a fall history in the last year. The area under the curve value of the model was 0.713. Risk factors of falls in PwMS included MSIS-29 (0.424), EDSS (0.406), marital status (0.297), education level (0.240), disease duration (0.185), age (0.130), family type (0.119), smoking (0.031), income level (0.031), and regular exercise habit (0.026). CONCLUSIONS: In this study, smoking and regular exercise were the modifiable factors contributing to falls in PwMS. We recommend that clinicians facilitate the modification of these factors in PwMS. Age and disease duration were non-modifiable factors. These should be considered as risk increasing factors and used to identify PwMS at risk. Interventions aimed at reducing MSIS-29 and EDSS scores will help to prevent falls in PwMS. Education of individuals to increase knowledge and awareness is recommended. Financial support policies for those with low income will help to reduce the risk of falls.
Asunto(s)
Accidentes por Caídas , Aprendizaje Automático , Esclerosis Múltiple , Humanos , Femenino , Masculino , Adulto , Estudios Transversales , Persona de Mediana Edad , Factores de RiesgoRESUMEN
Palbociclib is a dual inhibitor of cyclin-dependent kinases 4 and 6 (CDK4/6). Palbociclib has frequently been studied in breast cancer cells and has also been linked to function of P-glycoprotein (P-gp), main protein responsible for cancer drug resistance. However, the effect of Palbociclib on cancer drug resistance and specifically doxorubicin-resistant cells overexpressing P-gp have limitedly been studied in the literature. Here, we aimed to decipher the possible synergistic effects of Palbociclib and Doxorubicin combination treatment in doxorubicin-resistant not only breast cancer, which has restrictedly been studied previously, but leukemia and cervical cancer cell lines in the presence of sensitive counterparts to totally explore the mechanistic properties of the Palbociclib in cancer drug resistance. Our results underlined that Palbociclib differentially displayed synergistic effect with doxorubicin in a cell type-specific manner and increased the efficacy of Doxorubicin in Doxorubicin-resistant cells. As a monotherapy, palbociclib has been shown to decrease the expression of MDR-1 in doxorubicin-resistant cells, and when used in combination with doxorubicin, it has been shown to increase the accumulation of doxorubicin in the cell and consequently induce apoptosis. This is the first report that proposes the Palbociclib as a candidate for combination therapy to limit the Doxorubicin resistance in different cancer origins in clinics.
RESUMEN
Complex evolutionary dynamics governing the drug resistance is one of the major challenges in cancer treatment. Understanding these mechanisms requires a sequencing technology with higher resolution to delineate whether pre-existing or de novo drug mechanisms are behind the drug resistance. Combining this technology with clinically very relevant model system, namely 3D spheroids, better mimicking tumorigenesis and drug resistance have so far been lacking. Thus, we sought to establish dabrafenib and irinotecan resistant derivatives of barcoded 3D spheroids with the ultimate aim to quantify the selection-induced clonal dynamics and identify the genomic determinants in this model system. We found that dabrafenib and irinotecan induced drug resistance in 3D-HT-29 and 3D-HCT-116 spheroids are mediated by pre-existing and de novo resistant barcodes, indicating the presence of polyclonal drug resistance in this system. Moreover, whole-exome sequencing analysis found chromosomal gains and mutations associated with dabrafenib and irinotecan resistance in 3D-HT-29 and 3D-HCT-116 spheroids. Last, we show that dabrafenib and irinotecan resistance are also mediated by multiple drug resistance by detection of upregulation of the drug efflux pumps, ABCB1 and ABCG2, in our spheroid model system. Overall, we present the quantification of drug resistance and evolutionary dynamics in spheroids for the first time using cellular barcoding technology and the underlying genomic determinants of the drug resistance in our model system.
Asunto(s)
Esferoides Celulares , Tecnología , Humanos , Irinotecán/farmacología , Línea Celular Tumoral , Resistencia a MedicamentosRESUMEN
Heat shock response (HSR) which is regulated by heat shock factor 1 (HSF1) is the most important mechanism and the major regulator that prevents protein aggregation in neurodegenerative diseases. Excitotoxicity, which is the accumulation of excess glutamate in synaptic cleft, is observed in age-dependent neurodegenerative diseases and also in stroke, epilepsy, and brain trauma. Only a few studies in the literature show the link between excitotoxicity and HSR. In this study, we aimed to show the molecular mechanism underlying this link. We applied heat shock (HS) treatment and induced excitotoxicity with kainic acid (KA) in neuroblastoma (SHSY-5Y) and glia (immortalized human astrocytes (IHA)) cells. We observed that, only in SHSY-5Y cells, heat shock preconditioning increases cell survival after KA treatment. GLT-1 mRNA expression is increased as a result of KA treatment and HS due to the elevation of HSF1 binding to GLT-1 promoter which was induced by HSF1 phosphorylation and sumolation in SHSY-5Y cells. Additionally, glutamine synthetase and glutaminase expressions are increased after HS preconditioning in SHSY-5Y cells indicating that HS activates glutamate metabolism modulators and accelerates the glutamate cycle. In glia cells, we did not observe the effect of HS preconditioning. In summary, heat shock preconditioning might be protective against excitotoxicity-related cell death and degeneration.
Asunto(s)
Enfermedades Neurodegenerativas , Factores de Transcripción , Humanos , Factores de Transcripción/metabolismo , Proteínas de Unión al ADN/metabolismo , Factores de Transcripción del Choque Térmico/metabolismo , Ácido Kaínico/toxicidad , Respuesta al Choque Térmico/genética , Ácido Glutámico/toxicidadRESUMEN
INTRODUCTION: Placental inflammation is implicated in the pathophysiology of many pregnancy complications, including fetal growth restriction, preeclampsia, gestational diabetes, and choriocarcinoma. Mitochondrial dysfunction, one of the outcomes of placental inflammation, is characterized by loss of membrane potential, accumulation of oxygen radicals, mitochondrial protein folding defects, and disturbances in mitochondrial dynamics. Protein kinase R (PKR) is stimulated by double-stranded RNA and bacterial endotoxins in the presence of pathogens and is a critical immune response enzyme. PKR is also correlated with the cell death response during endoplasmic reticulum stress. In this study, we aim to investigate the effects of PKR activity stimulated by lipopolysaccharide (LPS), and double-stranded RNA analog (Poly I:C) on mitochondrial unfolded protein response (mtUPR), mitochondrial membrane potential, apoptosis, and oxidative stress in placental trophoblasts. METHODS: We applied LPS and Poly I:C to BeWo cells to induce PKR activation. In addition, cells were treated with 2-aminopurine (2-AP) to inhibit the kinase activity of PKR. Protein levels of ATP-dependent Clp protease proteolytic subunit (CLPP) and heat shock protein 60 (HSP60) were determined after treatments. Apoptotic markers were detected by real-time PCR and flow cytometry. PKR-induced reactive oxygen radicals (ROS) accumulation and mitochondrial membrane potential change were assessed by flow cytometry. RESULTS: It was determined that PKR activation-induced apoptosis in BeWo cells by reducing the levels of mtUPR proteins (CLPP and HSP60) and caused a decrease in mitochondrial membrane potential. PKR inhibition was sufficient for decreases in apoptotic markers and caused a reduction in the ratio of depolarized and ROS (+) cells. DISCUSSION: Our results showed that LPS and Poly I:C administration stimulated PKR in BeWo cells in vitro. Furthermore, PKR activation is correlated with the levels of proteins involved in mitochondrial homeostasis and apoptosis. Our findings will contribute to understanding the role of PKR activation in placental inflammation and related diseases.
Asunto(s)
Apoptosis , Inflamación , Placenta , Respuesta de Proteína Desplegada , eIF-2 Quinasa , Femenino , Humanos , Embarazo , eIF-2 Quinasa/genética , eIF-2 Quinasa/metabolismo , Inflamación/metabolismo , Lipopolisacáridos , Placenta/fisiopatología , Especies Reactivas de Oxígeno/metabolismo , ARN Bicatenario/metabolismo , Poli I-C/metabolismoRESUMEN
The removal of C. difficile inoculated on fresh spinach leaves washed with antimicrobial solutions was investigated. In addition, the effect of washing solutions on the total aerobic mesophilic bacteria (TAMB) and Enterobacteriaceae in the fresh spinach was examined. The fresh spinach was washed through immersion in different concentrations (MIC, 2xMIC, and 4xMIC) of the natural disinfectant solution (NDS) consisting of EDTA, borax, and epigallocatechin gallate (EGCG) content developed in our laboratory and green tea extract-acetic acid (GTE-AA) for varying contact times (5 and 15 min). Different concentrations (50, 100, and 200 ppm) of sodium hypochlorite (NaOCl) and tap water as the control group were used to compare the effectiveness of the NDS. In addition, the effects of washing on the color, texture, and total phenol content of the spinach were determined. No statistical difference was observed in the washing of the spinach leaves with NDS prepared at 2xMIC and 4xMIC concentrations, while inhibition of C. difficile ranged between 2.11 and 2.32 logs. The highest inhibition was observed in the application of 50 ppm NaOCl for 15 min with a decrease of 2.88 logs in C. difficile spores. The GTE-AA and NDS decreased the number of TAMB by 2.27-3.08 log and, 3.21-3.66 log, respectively. Washing spinach leaves with natural disinfectant for 5 min caused a decrease of 2.58 logs in Enterobacteriaceae load, while washing with 50 ppm NaOCl for 15 min reduced Enterobacteriaceae load by 4 logs. Tap water was ineffective in reducing any microbial load. No difference was detected in the color parameters of the spinach through all washes. Although all antimicrobial washes made a difference in the texture of the spinach, the greatest loss in firmness was observed in the spinach washed with NaOCl. Washing spinach with epigallocatechin-based wash solutions can remove C. difficile in possible C. difficile contamination, thereby reducing the environmental load of C. difficile. Epigallocatechin-based disinfectants can be an alternative to chlorine-based disinfectants in improving the microbial quality of vegetables.
Asunto(s)
Antiinfecciosos , Clostridioides difficile , Desinfectantes , Hipoclorito de Sodio/farmacología , Desinfección , Spinacia oleracea/microbiología , Clostridioides , Desinfectantes/farmacología , Ácido Acético/farmacología , Agua , Recuento de Colonia Microbiana , Microbiología de AlimentosRESUMEN
Commercial probiotic strains inhibited C. difficile, and other Clostridium cultures with zones 14.2-78.9 mm. The highest inhibition was observed with commercial culture on C. difficile ATCC 700057. Organic acids were the leading cause of inhibition. Probiotic cultures may be used as a support culture or through fermented foods for treatment.
Asunto(s)
Clostridioides difficile , Infecciones por Clostridium , Probióticos , Humanos , Clostridioides , Probióticos/farmacología , Clostridium , Infecciones por Clostridium/prevención & control , Antibacterianos/farmacologíaRESUMEN
Glial cells give rise to glioblastoma multiform as a primary brain tumor. In glioblastomas, neurons are destroyed via excitotoxicity which is the accumulation of excess glutamate in synaptic cavity. Glutamate Transporter 1 (GLT-1) is the main transporter that absorbs the excessive glutamate. Sirtuin 4 (SIRT4) was shown to have a potential protective role against excitotoxicity in previous studies. In this study, the regulation of dynamic GLT-1 expression by SIRT4 was analyzed in glia (immortalized human astrocytes) and glioblastoma (U87) cells. The expression of GLT-1 dimers and trimers were reduced and the ubiquitination of GLT-1 was increased in glioblastoma cells when SIRT4 was silenced; however GLT-1 monomer was not affected. In glia cells, SIRT4 reduction did not affect GLT-1 monomer, dimer, trimer expression or the ubiquitination of GLT-1. The phosphorylation of Nedd4-2 and the expression of PKC did not change in glioblastoma cells when SIRT4 was silenced but increased in glia cells. We also showed that SIRT4 deacetylates PKC in glia cells. In addition, GLT-1 was shown to be deacetylated by SIRT4 which might be a priority for ubiquitination. Therefore, we conclude that GLT-1 expression is regulated differently in glia and glioblastoma cells. SIRT4 activators or inhibitors of ubiquitination may be used to prevent excitotoxicity in glioblastomas.
Asunto(s)
Transportador 2 de Aminoácidos Excitadores , Glioblastoma , Sirtuinas , Humanos , Astrocitos/metabolismo , Transportador 2 de Aminoácidos Excitadores/genética , Glioblastoma/metabolismo , Ácido Glutámico/metabolismo , Proteínas Mitocondriales/metabolismo , Neuronas/metabolismo , Sirtuinas/metabolismo , Ubiquitinación , ProteolisisRESUMEN
Artificial Intelligence and algorithms are increasingly able to replace human workers in cognitively sophisticated tasks, including ones related to justice. Many governments and international organizations are discussing policies related to the application of algorithmic judges in courts. In this paper, we investigate the public perceptions of algorithmic judges. Across two experiments (N = 1,822), and an internal meta-analysis (N = 3,039), our results show that even though court users acknowledge several advantages of algorithms (i.e., cost and speed), they trust human judges more and have greater intentions to go to the court when a human (vs. an algorithmic) judge adjudicates. Additionally, we demonstrate that the extent that individuals trust algorithmic and human judges depends on the nature of the case: trust for algorithmic judges is especially low when legal cases involve emotional complexities (vs. technically complex or uncomplicated cases). Supplementary Information: The online version contains supplementary material available at 10.1007/s10506-022-09312-z.
RESUMEN
BACKGROUND: Cyanotic CHD is a life-threatening condition that presents with low oxygen saturation in the newborn period. Hypoxemia might cause alterations in the metabolic pathways. In the present study, we aimed to evaluate the early postnatal amino acid and carnitine/acylcarnitine profiles of newborn infants with cyanotic CHD. METHODS: A single centre case-control study was conducted. Twenty-seven patients with cyanotic CHD and 54 healthy newborn controls were enrolled. As part of the neonatal screening programme, results of amino acid and carnitine/acylcarnitine were recorded and compared between groups. RESULTS: Twenty-seven neonates with cyanotic CHD and 54 healthy newborns as controls were enrolled in the study. Cyanotic CHD neonates had higher levels of alanine, phenylalanine, leucine/isoleucine, citrulline, ornithine, C5, C5-OH; but lower levels of C3, C10, C12, C14, C14:1, C16, C16.1, C18, C5-DC, C6-DC, C16-OH, C16:1-OH when compared with the healthy controls. CONCLUSION: This study showed that there are differences between patients with cyanotic CHD and healthy controls in terms of postnatal amino acid and carnitine/acylcarnitine profiles.
Asunto(s)
Aminoácidos , Carnitina , Lactante , Humanos , Recién Nacido , Estudios de Casos y Controles , Carnitina/metabolismo , MetabolomaRESUMEN
BACKGROUND: Gestational diabetes mellitus (GDM) is a metabolic complication that affects millions of pregnant women in the world. Placental tissue function is endangered by hyperglycemia during GDM, which is correlated to increased incidences of pregnancy complications. Recently we showed that due to a significant decrease in mitochondrial fusion, mitochondrial dynamics equilibrium is altered in placental tissues from GDM patients. Evidence for the role of reduced mitochondrial fusion in the disruption of mitochondrial function in placental cells is limited. METHODS AND RESULTS: Here we show that chemical inhibition of mitochondrial fission in cultured placental trophoblast cells leads to an increase in mitochondrial fusion and improves the physiological state of these cells and hence, their capacity to cope in a hyperglycemic environment. Specifically, mitochondrial fission inhibition led to a reduction in reactive oxygen species (ROS) generation, mitochondrial unfolded protein marker expressions, and mitochondrial depolarization. It supported the increase in mitochondrial antioxidant enzyme expressions as well. Mitochondrial fission inhibition also increases the placental cell insulin sensitivity during hyperglycemia. CONCLUSION: Our results suggest that mitochondrial fusion/fission equilibrium is critical for placental cell function and signify the therapeutic potential of small molecule inhibitors of fission during GDM.
Asunto(s)
Diabetes Gestacional , Hiperglucemia , Embarazo , Femenino , Humanos , Placenta/metabolismo , Trofoblastos/metabolismo , Dinámicas Mitocondriales , Insulina/metabolismo , Diabetes Gestacional/metabolismo , Hiperglucemia/metabolismoRESUMEN
Antibiotic resistance genes can easily be transferred between bacteria in the biofilm. In the dairy industry, many bacterial species forming biofilms on the surfaces of equipment are widely reported. The experiments reported in this research paper aimed to investigate the carbapenem resistance and biofilm formation properties of Enterobacterales isolates which are spoilage microorganisms obtained from raw milk. In addition, the study determined that whether there was a relationship between the biofilm formation ability or the protein spectra of these isolates. In this study, ninety-two Enterobacterales isolates collected from 173 raw milk samples were investigated. Initially, the isolates were identified as Citrobacter braakii (n = 18), Citrobacter freundii (n = 12), Enterobacter asburiae (n = 1), Enterobacter cloacae (n = 3), Escherichia coli (n = 10), Hafnia alvei (n = 18), Klebsiella oxytoca (n = 1), Serratia fonticola (n = 24), Serratia liquefaciens (n = 4), and Serratia marcescens (n = 1) using MALDI-TOF MS. As a result, carbapenem resistance was determined in 6.5% of the isolates by CIM test, MHT, and the disk diffusion methods, but none of them had blaOXA-48, blaKPC, blaNDM-1, blaOXA23, blaOXA-58, blaOXA-51, blaVIM, and blaIMP genes. This may be due to the effect of other resistance mechanisms such as porin loss or increased flow pump activity. Furthermore, biofilm formation (weak and moderate) was detected in 97.8% of the Enterobacterales isolates. The mass spectra of the moderate biofilm producer isolate of Serratia spp. and the mass spectra of the weak biofilm producers of E.coli presented similarities.
Asunto(s)
Enterobacteriaceae , Leche , Animales , Enterobacteriaceae/genética , Leche/metabolismo , beta-Lactamasas/genética , beta-Lactamasas/metabolismo , Antibacterianos/farmacología , Escherichia coli/metabolismo , Carbapenémicos/farmacología , Biopelículas , Pruebas de Sensibilidad Microbiana , Proteínas Bacterianas/genéticaRESUMEN
Endoplasmic Reticulum (ER) stress is the response that occurs after the dysfunction of ER and its structure. Activated UPR triggers a stress response using ER membrane proteins such as PERK, IRE-1, GRP78, ATF5 ve ATF6. Sirtuins are enzymes that carry out post-translational modifications such as deacetylation and ADP-ribosylation. In our previous study, we identified Calreticulin as a SIRT4-interacting protein via mass spectrometry. Calreticulin binds to misfolded proteins, prevents them from leaving ER, which results in the reduction of ER stress. In this study, we aimed to investigate the interaction between SIRT4 and Calreticulin during ER stress in glia cells (IHA-immortalized human astrocytes). To trigger ER stress in glia cells, we first optimized the dose and the duration of the Tunicamycin which is 2.5 µg/ml concentration for 16 h. SIRT4 gene was silenced with lentiviral particles using 4 MOI (Multiplicity of Infection). In SIRT4-silenced cells, when treated with 2.5 µg/ml Tunicamycin for 16 h, the increase in the expressions of ATF6, GRP78 and the ratio of spliced/unspliced XBP1 mRNA were reduced. This shows that silencing SIRT4 may decrease ER stress. SIRT4-Calreticulin interaction was shown both in control and ER-stress induced glia cells. Additionally, this interaction did not change with the ER stress. SIRT4 only ADP-ribosylates Calreticulin during ER stress. Normally, SIRT4 ADP-ribosylates and deactivates Calreticulin during ER stress condition. When SIRT4 is silenced, the ADP-ribosylation level of Calreticulin decreases resulting in the activation of Calreticulin and the reduction of ER stress. In summary, SIRT4 inhibitors may be investigated as protective agents or drug candidates in neurodegenerative diseases where ER stress mostly underlies as one of the molecular mechanisms.
Asunto(s)
Astrocitos/metabolismo , Calreticulina/metabolismo , Estrés del Retículo Endoplásmico , Retículo Endoplásmico/metabolismo , Proteínas Mitocondriales/metabolismo , Sirtuinas/metabolismo , ADP-Ribosilación , Astrocitos/efectos de los fármacos , Línea Celular , Estrés del Retículo Endoplásmico/genética , Silenciador del Gen , Humanos , Proteínas Mitocondriales/antagonistas & inhibidores , Proteínas Mitocondriales/genética , Modelos Biológicos , Enfermedades Neurodegenerativas/tratamiento farmacológico , Sirtuinas/antagonistas & inhibidores , Sirtuinas/genética , Estrés Fisiológico/efectos de los fármacos , Tunicamicina/farmacologíaRESUMEN
BACKGROUND: The accumulation of excess glutamate in the synapse leads to excitotoxicity, which is the underlying reason of neuronal death in intracranial tumors. METHODS AND RESULTS: We identified the expression levels of glutamate dehydrogenase, glutamine synthetase and sirtuin 4 in U87 cell line and various intracranial tumors. mRNA expressions of glutamate dehydrogenase (GDH), glutamine synthetase (GS) and sirtuin 4 (SIRT4) were analyzed in various intracranial tumors using qPCR. GDH, GS and SIRT4 protein expressions were analyzed in glioblastoma (U87) and glial (IHA-immortalized human astrocytes) cell lines via western blotting. The protein expressions of SIRT4 and GS were shown to be elevated and GDH protein expression was reduced in U87 cells in comparison to IHA cells. All types of intracranial tumors displayed lower GS mRNA expressions compared to controls. SIRT4 mRNA expressions were also shown to be lower in all the tumors and grades, although not significantly. GDH mRNA expression was found to be similar in all groups. CONCLUSION: The molecular mechanisms of glutamate metabolism and excitotoxicity should be discovered to develop therapies against intracranial tumors.
Asunto(s)
Neoplasias Encefálicas/genética , Glioblastoma/genética , Adolescente , Adulto , Anciano , Astrocitos/metabolismo , Neoplasias Encefálicas/metabolismo , Línea Celular , Niño , Preescolar , Femenino , Expresión Génica/genética , Regulación Neoplásica de la Expresión Génica/genética , Glioblastoma/metabolismo , Glutamato Deshidrogenasa/genética , Glutamato-Amoníaco Ligasa/genética , Ácido Glutámico/metabolismo , Humanos , Masculino , Persona de Mediana Edad , Proteínas Mitocondriales/genética , Neuroglía/metabolismo , Estudios Retrospectivos , Sirtuinas/genéticaRESUMEN
INTRODUCTION: Gestational diabetes mellitus (GDM) poses a risk factor for fetal mortality and morbidity by directly affecting the placenta and fetus. Mitochondria are dynamic organelles that play a key role in energy production and conversion in placental tissue. Mitochondrial fusion and fission proteins are important in terms of providing mitochondrial dynamics, the adaptation of the cell to different conditions, and maintaining the metabolic stability of the cells. Although GDM shares many features with Type 2 diabetes mellitus (T2DM), different effects of these conditions on the mother and the child suggest that GDM may have specific pathological effects on placental cells. The aim of this study is to investigate the expression of mitochondrial dynamics, and mitochondrial protein folding markers in placentas from GDM patients and women with pre-existing diabetes mellitus. METHODS: Placentas were properly collected from women, who had pre-existing diabetes (Pre-DM), from women with gestational diabetes mellitus (GDM) and from healthy (non-diabetic) pregnant women. Levels of mitochondrial fusion markers were determined in these placentas by real time quantitative PCR and Western blot experiments. RESULTS: mRNA expressions and protein levels of mitochondrial fusion markers, mitofusin 1, mitofusin 2 (MFN1 and MFN2) and optical atrophy 1 (OPA1) proteins were found to be significantly lower in both Pre-DM placentas and those with GDM compared to healthy (non-diabetic) control group. Likewise, proteins involved in mitochondrial protein folding were also found to be significantly reduced compared to control group. DISCUSSION: Diabetes during pregnancy leads to processes that correlate with mitochondria dysfunction in placenta. Our results showed that mitochondrial fusion markers significantly decrease in placental tissue of women with GDM, compared to the healthy non-diabetic women. The decrease in mitochondrial fusion markers was more severe during GDM compared to the Pre-DM. Our results suggest that there may be differences in the pathophysiology of these conditions.
Asunto(s)
Diabetes Gestacional/metabolismo , Expresión Génica/fisiología , Dinámicas Mitocondriales/fisiología , Proteínas Mitocondriales/genética , Placenta/metabolismo , Adulto , Índice de Masa Corporal , Femenino , GTP Fosfohidrolasas/genética , Humanos , Proteínas de Transporte de Membrana Mitocondrial/genética , Obesidad/complicaciones , Obesidad/metabolismo , Embarazo , Complicaciones del Embarazo/metabolismo , ARN Mensajero/análisisRESUMEN
Glioblastoma multiform is a primary brain tumor derived from glial cells. The aim of this study is to investigate how glutamate metabolism is regulated by glutamate transporter 1 (GLT-1) degradation pathway in glioblastoma and glial cell lines. The protein expression levels of GLT-1, total ubiquitin, protein kinase C (PKC) proteins involved in the GLT-1 degradation pathway were measured by the western blot technique. Additionally, in glial and glioblastoma cells, the level of glutamate accumulated in the medium and the lysates was measured with the glutamate assay. GLT-1 protein expression was increased significantly in glioblastoma cells. The expression levels of the PKC protein and total ubiquitin were found to be decreased in glioblastoma cells although not significantly. The glutamate accumulated in the medium and lysates of glioblastoma cells is reduced compared to glial cells. Further research regarding excitotoxicity in glioblastoma focusing on GLT-1 degradation or activation pathway may create new opportunities of drug and treatment development.
Asunto(s)
Neoplasias Encefálicas/metabolismo , Transportador 2 de Aminoácidos Excitadores/metabolismo , Glioblastoma/metabolismo , Línea Celular Tumoral , Ácido Glutámico/metabolismo , Humanos , Proteolisis , Ubiquitina/metabolismoRESUMEN
INTRODUCTION: Plasma chemerin, which has chemotactic and adipogenic functions, is increased in several inflammatory diseases. However, its relationship with multiple sclerosis (MS) has not been explored yet. In this study, we aimed to determine chemerin levels and their possible role in MS. METHODS: Chemerin serum concentrations were evaluated by using ELISA kit in 91 clinically definite MS patients and 52 healthy controls. The mean serum chemerin, insulin, and cholesterol levels were compared. Patients were divided into two groups according to the body mass index (BMI), and the relationships between clinical and metabolic parameters were evaluated. RESULTS: Serum chemerin levels were 10.46 ± 1.65 ng/mL in MS patients and 10.26 ± 2.14 ng/mL in the control group. No significant difference was found between patients and controls (p = 0.55). We found no difference regarding age, gender, and BMI between two groups (p = 0.053, p = 0.54, p = 0.41). However, female patients with MS had higher chemerin levels than male patients. There were no associations between serum chemerin levels and EDSS score, annualized relapse rate, BMI, insulin resistance, and serum cholesterol levels in MS patients. CONCLUSION: In this study, we aimed to determine serum chemerin levels in patients with MS. However, in our study, there was no significant difference between serum chemerin levels of MS patients and healthy controls'. Additionally, chemerin levels were not associated with other metabolic parameters, as well as cognitive dysfunction. Further studies are needed to evaluate the role of chemerin in MS patients.
Asunto(s)
Resistencia a la Insulina , Esclerosis Múltiple , Índice de Masa Corporal , Quimiocinas , Femenino , Humanos , Péptidos y Proteínas de Señalización Intercelular , Lípidos , Masculino , ObesidadRESUMEN
PURPOSE: The purpose of our study was to investigate the mRNA expression profile of glutamate transporter 1 (GLT-1) in different types and grades of brain tumors, such as glioblastoma multiforme, astrocytomas (pilocytic, diffuse, anaplastic), oligodendrogliomas, ependydomas, medulloblastomas, and meningiomas using Real Time Quantitative PCR technique (qRT-PCR). METHODS: A total of 66 surgically removed primary brain tumors were collected retrospectively and the total RNA was isolated from each tumor sample. cDNA was generated and GLT-1 mRNA expression was evaluated with quantitative qRT-PCR. RESULTS: The mRNA expression of GLT-1 was significantly lower in primary brain tumors when compared to control brain tissues. GLT-1 expression was inversely correlated with the tumor grade, implicating its potential role in tumor progression. GLT-1 mRNA expression was lowest in grade 4 tumors, such as glioblastoma multiforme and medulloblastomas. The tumors with grade 3 and 4 combined displayed lower expression compared to tumors with grades 1 and 2. In grade 4 tumors, female patients displayed lower GLT-1 expression compared to male patients. In addition, glioblastoma multiforme patients older than 65 years of age showed lower GLT-1 expression when compared to the patients younger than 65. CONCLUSION: qRT-PCR was found to be a sensitive method in detecting GLT-1 expression in brain tumors. This study may lay the foundation for the future research about the excitotoxicity and brain tumors and GLT-1 might be a potential biomarker. Targeted therapies based on excitotoxic molecular pathways against gliomas should be designed to effectively combat these diseases.