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This study aimed to investigate the morphological, functional and molecular changes in frozen-thawed ram sperm using an extender containing different concentrations of hydrated carbon 60 fullerene (C60 HyFn), a nanotechnological product. Semen taken from each of the seven Akkaraman rams were pooled. Semen collection was done twice a week and it continued for 3 weeks. Each pooled semen sample was divided into six equal groups and diluted with tris + egg yolk extender including 0 (control), 200, 400, 800 nM, 1 and 5 µM concentrations of C60 HyFn at 37°C. They were then frozen in liquid nitrogen vapour at -140°C, stored in liquid nitrogen container (-196°C) and thawed at 37°C for 25 s before analysis. In comparison with control, C60 HyFn addition prior to freezing procedure provided significant increases in total and progressive motility rates, glutathione peroxidase, catalase activities and percentage of highly active mitochondria, and significant decreases in dead and abnormal sperm rates, lipid peroxidation, caspase-3 and DNA fragmentation levels in frozen-thawed ram semen. When compared to control, C60 HyFn supplementation significantly down-regulated the expression levels of miR-200a and KCNJ11, and significantly up-regulated the expression levels of miR-3958-3p (at the concentrations of 200, 400, 800 nM and 1 µM), CatSper1 (at the concentrations of 200, 400 nM and 5 µM), CatSper2 (at the concentrations of 1 and 5 µM), CatSper3 (at the concentrations of 200, 400 nM, 1 and 5 µM), CatSper4 (at all concentrations), ANO1 (at the concentrations of 800 nM, 1 and 5 µM) and TRPV5 (at the concentrations of 200, 400 and 800 nM). The addition of C60 HyFn had no effect on global DNA methylation rates. As a result, C60 HyFn supplementation to ram semen extenders may be beneficial in reducing some of the functional, structural and molecular damages in sperm induced by the freeze-thawing procedure.
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Fulerenos , MicroARNs , Preservación de Semen , Masculino , Ovinos , Animales , Semen , Fulerenos/farmacología , Motilidad Espermática , Crioprotectores/farmacología , Espermatozoides , Criopreservación/veterinaria , Criopreservación/métodos , Oveja Doméstica , Preservación de Semen/veterinaria , Preservación de Semen/métodos , Nitrógeno/farmacologíaRESUMEN
Carbamazepine (CBZ) is the antiepileptic drug used in epilepsy and some psychiatric disorders. Besides its widely used, many adverse effects have been reported including hematotoxicity, hepatotoxicity, endocrine disorders, and testicular damages due to oxidative stress. However, the role of CBZ on renal toxicity is not fully known. In this study, we attempted to explain the connected mechanisms by focusing on the metabolism of CBZ-induced renal toxicity in rats. Twenty male Wistar-Albino rats were randomized into 2 groups (n = 10); control (1 mL/day distilled water, orally) and CBZ (25 mg/kg/day CBZ, orally) groups. After 60 days, TAS (total oxidant status) and TOS (total oxidant status) levels, histopathological features, some genes involved in apoptosis, 8-hydroxy-2-deoxyguanosine (8-OHdG) activity, and apoptotic cells were assessed of kidney tissue. The oxidative stress index (OSI) was measured from TAS and TOS levels. TOS levels and OSI significantly increased, while TAS levels decreased in the CBZ group relative to the control group. Histopathological observations, Caspase-3 (Casp3), Poly [ADP-ribose] polymerase-1 (PARP-1), 8-OHdG immunoreactivities, and apoptotic cells markedly raised in the CBZ group compared with the control group. Also, mRNA expression of Cytochrome c (Cytc) and CASP3 significantly increased in the CBZ group compared to the control group. In conclusion, long-term use of CBZ may promote renal damage in rats by inducing oxidative stress and apoptosis.
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Apoptosis , Estrés Oxidativo , Animales , Ratas , Masculino , Caspasa 3 , Ratas Wistar , Carbamazepina/toxicidad , 8-Hidroxi-2'-Desoxicoguanosina/farmacología , OxidantesRESUMEN
PURPOSE: Vestigial like family member 3 (VGLL3) and its sub-target genes show considerable transcriptomic overlap in terms of several autoimmune and inflammatory diseases. Herein, we investigated the role of VGLL3 rs13074432 polymorphism and its sub-target genes in the aetiology of acute appendicitis (AA). METHODS: In this prospective case-control study, we included 250 patients (age, 0-18 years) who underwent appendectomy with the diagnosis of AA (patient group; blood and appendix tissue samples) and 200 healthy children (control group; only blood samples) without appendectomy. ELISA method was used for protein-level detection of VGLL3 and sub-target genes expression change in obtained tissue samples, and real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used for mRNA level detection. Genotyping analyses were performed on DNA samples isolated from blood using TaqMan SNP genotyping test. RESULTS: The frequency of TT variant genotype (p < 0.001) and T allele (p = 0.002) showed a significant decrease in the patient group compared with the control group. No significant correlation was observed between the expression of VGLL3 in the appendiceal tissue and patient clinical and demographic data (p > 0.050). CONCLUSION: This study revealed that the VGLL3 gene and its sub-target genes are associated with AA aetiology.
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Apendicitis , Apéndice , Humanos , Niño , Recién Nacido , Lactante , Preescolar , Adolescente , Apendicitis/genética , Apendicitis/cirugía , Apendicitis/diagnóstico , Estudios de Casos y Controles , Apendicectomía , ADN , Enfermedad Aguda , Factores de TranscripciónRESUMEN
Background/aim: By applying humanin (HN) before myocardial infarction (MI), its protection in myocardial injury and the possible roles of its cellular mechanism in the Notch pathway were investigated. Materials and methods: The study was carried out at Firat University Experimental Research Center (12/24/2018-12/23/2019). Spraque-Dawley rats were divided into 10 groups: I (control) (n = 6), II (HN 6 h) (n = 6), III (HN 24 h) (n = 6), IV (HN day 7) (n = 6), V (MI 6 h) (n = 7), VI (MI 24 h) (n = 7), VII (MI day 7) (n = 7), VIII (MI+HN 6 h) (n = 7), IX (MI+HN 24 h) (n = 7), and X (MI+HN day 7) (n = 7). To create MI, 200 mg/kg of isoproterenol (ISO) was administered to the rats subcutaneously. Moreover, 252 µg/kg of HN was given intraperitoneally (ip) to the rats on its own and before MI. Molecular parameters Notch1, Notch2, Hes1, Hes2, Jagged1, Jagged2, DLL1, and DLL4 were examined using polymerase chain reaction in the heart tissue, Notch1, Hes1, and DLL4 were examined using western blot, while heart tissue was taken for histochemical examinations. Results: The mRNA expression levels of the Notch signaling members (Notch1, Notch2, Hes1, Hes2, Jagged1, Jagged2, DLL1, and DLL4) tended to decrease after MI. The Notch signaling members increased more significantly, especially toward day 7 after HN application before MI. In the western blot anylyses, the Notch1, Hes1, and DLL4 protein levels increased significantly toward day 7 in the groups given HN before MI. Moreover, the serum AST, LDH, CK-MB, and troponin I levels tended to decrease with the application of HN before MI and there was a significant decrease in edema, hemorrhage, and mononuclear cells in the heart tissue at 24 h post-MI and fibrosis on day 7 post-MI. Conclusion: HN administration before MI has a cardioprotective effect on rats via the Notch signaling pathway.
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Infarto del Miocardio , Ratas Sprague-Dawley , Transducción de Señal , Animales , Infarto del Miocardio/metabolismo , Transducción de Señal/efectos de los fármacos , Ratas , Masculino , Receptores Notch/metabolismo , Péptidos y Proteínas de Señalización Intercelular/metabolismo , Péptidos y Proteínas de Señalización Intracelular/metabolismo , Péptidos y Proteínas de Señalización Intracelular/genética , Modelos Animales de EnfermedadRESUMEN
This study was carried out to investigate the effect of the semen freeze-thawing process on the functionality and molecular structure of ram spermatozoa. The temperature of pooled and diluted semen at 38°C (group 1, control) was lowered to 5°C (group 2), and it was subjected to glycerolisation-equilibration (group 3), frozen and thawed (group 4). Compared to the control, deterioration in spermatological parameters and significant increases in lipid peroxidation and global DNA methylation levels were observed in groups 3 and 4. When compared with the control, significant downregulation in the levels of miR-485 of group 2, miR-29a of group 3 and let-7a, miR-485 and miR-29a of group 4, and significant upregulation in the levels of miR-107 of group 3 and miR-127 of groups 3 and 4 were detected. In comparison to the control, significant upregulation in the levels of CatSper1, CatSper2, CatSper3, CatSper4, ANO1 and TRPM3 of group 2, CatSper4, ANO1 and TRPM3 of group 3 and KCNJ11 of group 4, and significant downregulation in the CatSper 3 level of group 4 were determined. As a result, the semen freeze-thawing process causes motility and morphological disorders in rams. This may be due to molecular changes associated with lipid peroxidation in spermatozoa.
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Criopreservación/veterinaria , Metilación de ADN/fisiología , Peroxidación de Lípido/fisiología , Preservación de Semen/veterinaria , Ovinos , Espermatozoides/fisiología , Animales , Apoptosis , Regulación de la Expresión Génica/fisiología , Calor , Canales Iónicos/genética , Masculino , MicroARNs/genética , Estrés Oxidativo , Preservación de Semen/efectos adversos , Motilidad EspermáticaRESUMEN
Male infertility is a global health problem, and the underlying molecular mechanisms are not clearly known. Ion channels and microRNAs (miRNAs), known to function in many vital functions in cells, have been shown to play a significant role in male infertility through changes in their expressions. The study aimed to evaluate the alterations of testicular and/or spermatozoal potassium voltage-gated channel subfamily J member 11 (KCNJ11), Cystic fibrosis transmembrane conductance regulator (CFTR), miR-let-7a and miR-27a expressions in carbamazepine-related male infertility. Here, we showed that carbamazepine reduced sperm motility, increased abnormal sperm morphology, and impaired hormonal balance as well as increased relative testis weight and decreased relative seminal vesicle weight. On the other hand, downregulated KCNJ11 and upregulated miR-let-7a expressions were determined in testis (p < .05). Also, downregulated KCNJ11 and upregulated CFTR and miR-27a expressions were found in spermatozoa (p < .05). Interestingly, altered testicular KCNJ11 and miR-let-7a expressions were correlated with decreased sperm motility and elevated sperm tail defect. Besides, spermatozoal CFTR and miR-27a expressions positively correlated with sperm tail defects. The results indicated a significant relationship between ion channel and/or miRNA expression alterations and impaired sperm parameters due to carbamazepine usage.
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MicroARNs , Motilidad Espermática , Carbamazepina/efectos adversos , Regulador de Conductancia de Transmembrana de Fibrosis Quística/genética , Humanos , Masculino , MicroARNs/genética , Espermatozoides , TestículoRESUMEN
BACKGROUND: The COVID-19 pandemic is a unique challenge to the care of patients with hematological malignancies. We aim to provide supportive guidance to clinicians making individual patients decisions during the COVID-19 pandemic, in particular during periods that access to healthcare resources may be limited. DISCUSSION: This review also provides recommendations, which are convenient in evaluating indications for therapy, reducing therapy-associated immunosuppression, and reducing healthcare utilization in patients with specific hematological malignancies in the COVID-19 era. Specific decisions regarding treatment of hematological malignancies will need to be individualized, based on disease risk, risk of immunosuppression, rates of community transmission of SARS-CoV-2, and available local healthcare resources.
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COVID-19 , Neoplasias Hematológicas , Neoplasias Hematológicas/epidemiología , Neoplasias Hematológicas/terapia , Humanos , Pandemias/prevención & control , SARS-CoV-2RESUMEN
The effect of verapamil, a calcium channel blocker, on male fertility in terms of ion channel and miRNA gene expressions in testis/spermatozoa was evaluated in this study. Rats were divided into sham and verapamil groups (n = 15). Verapamil was performed orally for 60 days. Sperm parameters and levels of serum follicle-stimulating hormone (FSH), luteinising hormone (LH) and testosterone (T) hormones were analysed. Alterations of microRNA (miRNA) and ion channel gene expressions in spermatozoa/testis were detected by using qPCR. Verapamil treatment reduced sperm concentration. Increased serum FSH, LH and T hormone levels were detected. Upregulated transient receptor potential cation channel subfamily V member 5 (TRPV5) and potassium voltage-gated channel subfamily J member 11 (KCNJ11) gene expressions and downregulated miR-let-7b, miR-10a, miR-320 and miR-760 expressions were found in testis of verapamil group. However, upregulated anoctamin 1 (ANO1), ATP-binding cassette subfamily C member 9 (ABCC9), miR-27a and miR-130a expressions and downregulated miR-20a, miR-92a, miR-132, miR-320 and miR-760 expressions were detected in spermatozoa. In addition, these altered gene expressions were found to be associated with decreased sperm concentration. The results indicate that the changes in testicular and/or spermatozoal ion channels and miRNA expressions due to verapamil treatment may affect male fertility.
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Infertilidad Masculina , MicroARNs , Animales , Hormona Folículo Estimulante , Humanos , Infertilidad Masculina/inducido químicamente , Infertilidad Masculina/genética , Canales Iónicos , Masculino , MicroARNs/genética , Ratas , Espermatozoides , Testículo , Testosterona , Verapamilo/farmacologíaRESUMEN
INTRODUCTION: The impaired balance between cell proliferation and cell death, followed the inability to receive the death signals, cells push towards the neoplasia pathway. Fibulin 1 (FBLN1) plays a role as a co-factor in the mechanism of action of a protease such as a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS-1), which has important roles in angiogenesis, can also act as both tumor suppressor gene (TSG) and an oncogene in the main constituent of the extra-cellular matrix. This preliminary study has investigated the effects of silencing FBLN1 with siRNA on autophagy, proliferation, apoptosis pathways in the MSM cell line. MATERIAL AND METHODS: It was transfected siRNA specific to FBLN1 incubated MSM SPC212 cells, and compared with negative control siRNAs by a real-time polymerase chain reaction. It was determined apoptosis, proliferation, autophagy-related genes in mRNA levels. RESULTS: It was observed that increased anti-apoptosis genes, such as CASP2, CASP7, DDFA, and BCL2, anti-apoptotic gene, reduced APAF1, CASP8. Proliferation induced through while increased ADAMTS1, CDH1, CDH6, CLDN7, CSF3, MMP7, MMP13 genes. Autophagy increased via increasing MAP1LC3B, ATG-16L1 genes while decreased via suppressed ULK1, and ATG7 genes by silencing FBLN1 with siRNAs (p < 0.05). CONCLUSIONS: Proliferation can be induction with silencing of FBLN1 with siRNA in processing mechanism MSM. It was concluded that FBLN1 could be act as pleiotropic on autophagy, and apoptosis pathways in proliferation processing for MSM. Therefore we think that FBLN1 acts like a TSG. FBLN1 can be considered as a targeted treatment option in advanced stage MSM.
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Acid-sensing ion channels (ASICs) are voltage-independent and proton-gated channels. In this study, we aimed to test the hypothesis whether ASICs might be involved in modifying the excitability of stellate cells in the cochlear nucleus (CN). We determined gene expressions of ASIC1, ASIC2 and ASIC3 in the CN of BALB/mice. ASIC currents in stellate cells were characterized by using whole-cell patch-clamp technique. In the voltage-clamp experiments, inward currents were recorded upon application of 2-[N-Morpholino ethanesulfonic acid]-normal artificial cerebrospinal fluid (MES-aCSF), whose pH 50 was 5.84. Amiloride inhibited the acid-induced currents in a dose-dependent manner. Inhibition of the ASIC currents by extracellular Ca2+ and Pb2+ (10 µM) was significant evidence for the existence of homomeric ASIC1a subunits. ASIC currents were increased by 20% upon extracellular application of Zn2+ (300 µM) (p < 0.05, n = 13). In current-clamp experiments, application of MES-aCSF resulted in the depolarization of stellate cells. The results show that the ASIC currents in stellate cells of the cochlear nucleus are carried largely by the ASIC1a and ASIC2a channels. ASIC channels affect the excitability of the stellate cells and therefore they appear to have a role in the processing of auditory information.
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Canales Iónicos Sensibles al Ácido/metabolismo , Percepción Auditiva/fisiología , Núcleo Coclear/fisiología , Neuronas/fisiología , Animales , Ratones , Ratones Endogámicos BALB CRESUMEN
Background/aim: This study compared TRPM2 and TRPM7 ion channel gene expression and immunohistochemical staining in endometrial hyperplasia and endometrium adenocarcinoma. Materials and methods: Sections were taken from paraffin blocks of 120 patients who were divided into 6 groups as follows: G1 (n = 20), proliferative endometrium (PE); G2 (n = 20), EH without atypia; G3 (n = 20), EH with atypia; G4 (n = 20), stage 1A, grade 1 EC; G5 (n = 20), stage 1A, grade 2 EC; and G6 (n = 20), stage 1A, grade 3 EC. TRPM2 and TRPM7 genes were analyzed with qRT-PCR in paraffin-embedded tissue samples. Under light microscopy, TRPM2 and TRPM7 immunostaining scores of the samples taken from polylysine slides were evaluated. Results: Compared to G1, TRPM2 mRNA gene expression was significantly downregulated in G3 and G5. TRPM2 immunoreactivity scores were similar in all groups. TRPM7 mRNA gene expression was significantly downregulated in G2, G3, and G6 when compared to G1. TRPM7 immunoreactivity scores were similar in G1, G2, and G3, but significantly decreased in G4, G5, and G6 Conclusion: Reduction in TRPM7 ion channel activity may be a progression marker for endometrial hyperplasia regardless of the atypical criteria.
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Hiperplasia Endometrial/metabolismo , Neoplasias Endometriales/metabolismo , Proteínas Serina-Treonina Quinasas/metabolismo , Canales Catiónicos TRPM/metabolismo , Biomarcadores/metabolismo , Hiperplasia Endometrial/fisiopatología , Neoplasias Endometriales/fisiopatología , Femenino , Regulación de la Expresión Génica , Humanos , Inmunohistoquímica , Proteínas Serina-Treonina Quinasas/genética , Estudios Retrospectivos , Canales Catiónicos TRPM/genéticaRESUMEN
Background/aim: Insulin-like growth factor (IGF)-I is a differentiation and growth factor. Antifibrotic action of octreotide has been reported in pulmonary fibrosis. The present study aimed to research the prophylactic and therapeutic potential of octreotide on a bleomycin (BLM)-induced experimental scleroderma model. Materials and methods: Sixty Balb/c female mice were divided into 6 groups. Daily subcutaneous BLM (100 µg) was injected for 3 weeks in groups II and III and for 6 weeks in groups V and VI. Octreotide (100 µg/kg per day) was injected subcutaneously for the first 3 weeks in group III (prophylactic) and the second 3 weeks in group VI (therapeutic). Mice in groups I, II, and III were sacrificed at the end of the third week, while mice in groups IV, V, and VI were sacrificed at the end of the sixth week. Results: Repeated BLM applications increased dermal inflammatory cell counts and dermal thickness, and led to dermal fibrosis at both the third and sixth weeks. Moreover, mRNA expressions of TGF-ß1 and IGF binding protein (IGFBP)-3 and -5 were higher in the BLM- injected sham groups. On the other hand, IGFBP-3 and -5 mRNA expressions were significantly decreased in both the prophylactic and therapeutic octreotide groups. Similarly, octreotide decreased dermal inflammatory infiltrations and dermal thickness. Conclusion: Octreotide has antifibrotic actions on experimentally induced dermal fibrosis. It can be suggested that IGF-I plays pathogenic roles, and octreotide is a candidate for research in the treatment of scleroderma.
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Antineoplásicos Hormonales/uso terapéutico , Factor I del Crecimiento Similar a la Insulina/metabolismo , Octreótido/uso terapéutico , Esclerodermia Localizada/tratamiento farmacológico , Piel/efectos de los fármacos , Animales , Antineoplásicos Hormonales/farmacología , Bleomicina , Modelos Animales de Enfermedad , Femenino , Fibrosis/tratamiento farmacológico , Fibrosis/metabolismo , Inflamación/tratamiento farmacológico , Inflamación/metabolismo , Inyecciones Subcutáneas , Proteínas de Unión a Factor de Crecimiento Similar a la Insulina/genética , Proteínas de Unión a Factor de Crecimiento Similar a la Insulina/metabolismo , Ratones Endogámicos BALB C , Octreótido/farmacología , ARN Mensajero/metabolismo , Esclerodermia Localizada/inducido químicamente , Esclerodermia Localizada/patología , Piel/metabolismo , Piel/patología , Factor de Crecimiento Transformador beta1/genética , Factor de Crecimiento Transformador beta1/metabolismoRESUMEN
BACKGROUND: To investigate the mechanism of action of stathmin1 (STMN1) in mesothelioma (MSM) and whether it has any role in its treatment. METHODS: STMN1 expression was examined using immunohistochemistry in biopsy tissues taken from MSM patients. The relationships between the levels of STMN1 expression in the pathology preparations of MSM patients, and the clinicopathological characteristics of these patients, and their survival times were investigated. Transfection of STMN1-specific siRNA into SPC212 cells was compared to negative control siRNAs. The mRNA levels of genes that may play a role in invasion, apoptosis, and autophagy were evaluated by RT-PCR. RESULTS: The expression of STMN1 was shown to be high in MSM tissues (p < 0.05). It was found that the only independent predictor factor affecting the survival time of MSM patients was the disease stage (p < 0.05). STMN1 was significantly reduced after siRNA intervention (81.5%). STMN1 with specific siRNA has been shown to suppress invasion by reducing the mRNA levels of cadherin-6 (CDH6), fibroblast growth factor-8 (FGF8), hypoxia-inducible factor 1 (HIF1A), matrix metallopeptidase 1-2 (gelatinase A) (MMP1-2), and TIMP metallopeptidase inhibitor 2 (TIMP2), which are important markers for invasion. Although the expression of apoptosis and autophagy-related genes, caspase-2 (Casp2) and LC-3, was reduced by silencing STMN1 with specific siRNA in western blot analysis, this effect was not observed in PCR results. CONCLUSIONS: Immunohistochemical analysis of STMN1 may contribute to the differential diagnosis of MSM, and STMN1 may also be considered as a potential therapeutic target in the early invasive stage of MSM therapy.
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Mesotelioma Maligno , Mesotelioma , Humanos , Mesotelioma/genética , Metaloproteasas , ARN Mensajero , ARN Interferente Pequeño/genética , Estatmina/genética , Estatmina/metabolismoRESUMEN
OBJECTIVE: The aim of our study was to investigate the relationship between the destruction of temporal bone structures, ossicular chain destruction, dissemination of cholesteatoma and the expressions of bone morphogenetic proteins (BMPs), BMP-2, BMP-4 and BMP-6 in patients with acquired cholesteatoma. MATERIAL AND METHODS: This study was performed in a total of 80 patients with cholesteatoma and without cholesteatoma who had undergone surgery due to chronic otitis media. The patients were grouped as the study and the control groups. The study group comprised patients with primary acquired cholesteatoma, and the control group consisted of chronic otitis media patients without cholesteatoma. The samples were obtained from cholesteatoma tissue and the external acoustic meatus skin in study group patients and they were obtained from the external acoustic meatus skin only in control group patients. The Reverse Transcriptase Polymerase Chain Reaction method was used for the measurements of BMPs, BMP-2, BMP-4 and BMP-6 expressions. Polymerase Chain Reaction was studied by isolation of Ribonucleic Acid from the tissue samples. RESULTS: When the expressions of BMP in the external acoustic meatus skin were compared between the study and the control groups, the BMPs, BMP-2 and BMP-6 were determined to have a statistically significant relation in the study group (p<0.05), but BMP-4 was not significant (p>0.05). When the expression of BMP in cholesteatoma tissue was investigated in the study group patients, the BMPs, BMP-2 and BMP-6 were determined with statistically significant positivity (p<0.05), but there was no significant positivity for BMP-4 (p>0.05). In the study group, there was no statistical significance between the expressions of BMPs, BMP-2, BMP-4 and BMP-6 in cholesteatoma tissue, in the external acoustic meatus skin, and temporal and ossicular chain destruction, and dissemination of cholesteatoma (p>0.05). A statistically significant positivity for BMPs expression in cholesteatoma tissue was determined in patients with destruction of the incus+malleus+stapes (p<0.05). CONCLUSION: The expressions of BMPs, BMP-2 and BMP-6, were elevated in cholesteatoma tissue. Furthermore, the positivity of BMPs expression was statistically significant in patients with destruction of all the ossicles, and we think that this marker can be used for evaluation of the aggressiveness of cholesteatoma.
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Proteínas Morfogenéticas Óseas/metabolismo , Colesteatoma del Oído Medio/metabolismo , Colesteatoma del Oído Medio/patología , Adolescente , Adulto , Proteínas Morfogenéticas Óseas/genética , Estudios de Casos y Controles , Conducto Auditivo Externo/metabolismo , Osículos del Oído/patología , Electroforesis en Gel de Agar , Femenino , Humanos , Masculino , Persona de Mediana Edad , Otitis Media/patología , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Piel/metabolismo , Adulto JovenRESUMEN
BACKGROUND: Mutual regulation between immune system and gut microbiota is achieved through several mechanisms including the engagement of toll-like receptors (TLRs) which is expressed on numerous cell types. In this study we aimed to explore the association between food allergies and TLR gene polymorphisms in association with gut microbiota. METHODS: Toll-like receptors polymorphism frequencies and some bacteria in the gut microbiota in 130 infants aged 1-24 months with egg and/or milk allergy in a prospective cohort were compared with 110 non-food allergic controls. Four candidate polymorphisms (TLR2 rs1898830/rs5743708 and TLR4 rs4986790/rs4986791) were genotyped by allelic discrimination polymerase chain rection (PCR) method. Gut microbiota analysis was achieved by using high-throughput sequencing. RESULTS: The TLR4 rs4986790 (Asp299Gly) single nucleotide polymorphism (SNP) major/minor allele frequency was 0.788/0.212 in food allergy patients and 0.719/0.280 in controls (p=0.017). There was a statistically significant difference between groups in terms of genotype frequencies (AA, AG, GG). Gut microbiota analysis revealed increased Firmicutes phylum in stool of the patients with food allergy. Except for TLR4 rs4986791 (Thr399lle) allele, the other TLR polymorphisms were not associated with food allergies in children. When the bacteria in the intestinal microbiota and TLR2 and TLR4 gene polymorphisms were compared; we determined a statistically significant increase in Bifidobacterium concentration in the intestinal microbiota in TLR4 rs4986791 CT heterozygous genotype (p=0.004). CONCLUSIONS: This study demonstrated a partial role of TLR4 gene polymorphism and gut microbiota in the development of food allergies. Future work in this area will be required to clarify the roles of different microbial strains that modulate gut microbiota composition and function in conjunction with TLR transcription pathways.
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Hipersensibilidad a los Alimentos , Microbioma Gastrointestinal , Humanos , Niño , Predisposición Genética a la Enfermedad , Receptor Toll-Like 2/genética , Receptor Toll-Like 4/genética , Receptor Toll-Like 4/metabolismo , Estudios Prospectivos , Polimorfismo de Nucleótido Simple , Hipersensibilidad a los Alimentos/genética , Estudios de Casos y ControlesRESUMEN
INTRODUCTION: Diabetes mellitus (DM) is a common, chronic metabolic disease that has harmful effects on many diverse tissues, including the testis. One of the ways of tissue damage is the modification of transient receptor potential melastatin 2 (TRPM2) channels by increased reactive oxygen species (ROS). In our study for the first time, it was aimed to investigate TRPM2 channel activation in testicular tissues of diabetic rats induced by streptozotosin (STZ) and to examine the efficacy of N-acetylcysteine (NAC) treatment, which is an antioxidant. METHODS: In our study, 28 Wistar albino male rats aged 8-10 weeks were used, and animals were divided into four groups: control group, NAC group, DM group, and DM + NAC group. The experimental phase was designed as eight weeks. The malondialdehyde (MDA) level, which is an indicator for lipid peroxidation due to oxidative stress, was measured by the spectrophotometric method. The Tunel assay was used to determine apoptosis on testicular tissue. TRPM2 immunoreactivity was determined by the avidin-biotin-peroxidase complex method, and quantitative polymerase chain reaction (QPCR) was used to determine TRPM2 expression levels. RESULTS: It was seen that MDA levels were significantly increased in the DM group and decreased after NAC treatment. Similarly, it was observed that apoptosis levels, which increased significantly in diabetic rats, decreased to the levels of the control group after treatment. It was seen that TRPM2 activation and expression levels were significantly decreased in the DM group. CONCLUSION: The results of this study show that NAC regulates TRPM2 activation in the testicular tissue of patients with diabetes and has tissue-protective properties.
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This study was conducted to determine the possible effects of intracerebroventricular MOTS-c infusion on thyroid hormones and uncoupling proteins (UCPs) in rats. Forty male Wistar Albino rats were divided into 4 groups with 10 animals in each group: control, sham, 10 and 100 µM MOTS-c. Hypothalamus, blood, muscle, adipose tissues samples were collected for thyrotropin-releasing hormone (TRH), UCP1 and UCP3 levels were determined by the RT-PCR and western blot analysis. Serum thyroid hormone levels were determined by the ELISA assays. MOTS-c infusion was found to increase food consumption but it did not cause any changes in the body weight. MOTS-c decreased serum TSH, T3, and T4 hormone levels. On the other hand, it was also found that MOTS-c administration increased UCP1 and UCP3 levels in peripheral tissues. The findings obtained in the study show that central MOTS-c infusion is a directly effective agent in energy metabolism.
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Hormonas Tiroideas , Ratas , Masculino , Animales , Proteínas Desacopladoras Mitocondriales , Ratas Wistar , Hormonas Tiroideas/farmacología , Peso CorporalRESUMEN
Stathmin1 (STMN1) has been proposed as a possible prognostic marker and a potential therapeutic target for some cancers. We aimed to analyze the changes in autophagy, invasion, apoptosis-related genes in prostate cancer (PCa) cell line (PC-3), after small interfering RNA (siRNA)-mediated STMN1 silencing, and also the relationships of STMN1 expression, clinicopathological parameters, and survival (OS) in PCa cases. The STMN1 expressions were analyzed, immunohistochemically, in formalin-fixed paraffin-embedded 75 PCa and 15 benign prostatic hypertrophy (BPH) tissues. The correlation between the levels of expression STMN1, clinicopathological features, and OS was determined in PCa cases. The siRNA-mediated STMN1 incubated PC-3 cells were transfected and compared to negative control siRNAs. We determined mRNA levels in autophagy, invasion, and apoptosis genes with the combination of reverse transcription-polymerase chain reaction (RT-PCR) and western blotting in PC3 cell lines after STMN1 silencing. It was determined that STMN1 was overexpressed significantly in PCa cases, immunohistochemically. The overexpression of STMN1 was significantly correlated with the high-grade Gleason score, and it was associated with a worse prognosis of PCa cases according to the Kaplan-Meier survival analysis (p < 0.05). Significant silencing in STMN1 was determined (87.5%) after siRNA applications. Especially, invasion genes such as claudin 7, fibroblast growth factor 8, hypoxia-inducible factor 1 subunit alpha, hepatocyte growth factor, matrix metallopeptidase 2, 7 genes, markedly, decreased by siRNA-mediated STMN1silencing. STMN1 silencing was determined to significantly increase caspase 3 protein expression by using western blot analysis (p < 0.001). Although STMN1 silencing did not have a significant effect on the induction of apoptosis and autophagy-related genes in PCa cells, it was shown to affect apoptotic mechanisms through the caps3 protein. siRNA-mediated STMN1 silencing decreases proliferation in the PCa cell line. It is thought that STMN1 can serve as a potential therapeutic target in the advanced stage-PCa, especially.
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OBJECTIVE: To investigate whether m6A content changes in type 2 diabetes mellitus (T2DM) and obese individuals and whether the relationship of m6A content with the mRNA expression levels of FTO and ALKBH5 genes. STUDY DESIGN: Cross-sectional study. PLACE AND DURATION OF STUDY: Department of Internal Medicine, Firat University, Medical School, Elazig, Turkey, between January 2019 and January 2022. METHODOLOGY: The study included 34 newly diagnosed patients with type 2 diabetes mellitus, 34 obese individuals, and 33 healthy individuals without any chronic and metabolic disease matched for age and gender. The global m6A RNA methylation, FTO, and ALKBH5 gene analyses of all the participants were performed. Total cholesterol, triglyceride, LDL, HDL, HbA1c, and insulin and glucose levels were measured. RESULTS: The median percentages of m6A RNA methylation in the control group, obese, and T2DM participants were 5.62%, 4.20%, and 5.21% respectively (p=0.004). The m6A RNA methylation percentage of the obese participants was significantly lower than controls (p=0.021). The FTO and ALKBH5 mRNA levels were significantly lower in obese and T2DM participants than in controls. There was a negative significant correlation between m6A RNA level and FTO i.e. (r=-0.291, p=0.003) and ALKBH5 (r=-0.321. p=0.001) levels. CONCLUSION: m6A RNA expression levels of obese individuals were lower than healthy controls. The FTO and ALKBH5 mRNA expressions were lower in both obese and T2DM participants compared to the healthy controls. There was no significant difference between obese and T2DM individuals in terms of m6A RNA expression, FTO and ALKBH5 mRNA expression. m6A RNA expression, FTO, and ALKBH5 levels have a potential role in obesity and diabetes mellitus. KEY WORDS: m6A RNA methylation, Epigenesis, Genetic, FTO, ALKBH5.
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Diabetes Mellitus Tipo 2 , ARN , Adenosina/metabolismo , Desmetilasa de ARN, Homólogo 5 de AlkB/genética , Desmetilasa de ARN, Homólogo 5 de AlkB/metabolismo , Dioxigenasa FTO Dependiente de Alfa-Cetoglutarato/genética , Dioxigenasa FTO Dependiente de Alfa-Cetoglutarato/metabolismo , Estudios Transversales , Diabetes Mellitus Tipo 2/genética , Humanos , Obesidad/genética , ARN/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
OBJECTIVE: To determine the difference in serum Elabela level in hypertensive patients with and without nephropathy compared to the healthy control group. Study Desing: Cross-sectional descriptive study. PLACE AND DURATION OF STUDY: Firat University Medical School, Elazig, Turkey between December 2018 and November 2020. METHODOLOGY: The cross-sectional descriptive study consisted of 37 patients with hypertensive nephropathy (group 3), 50 hypertensive patients without nephropathy (group 2), and 50 healthy controls (group 1). Hypertensive nephropathy was defined as serum creatinine ≥1.8 mg / dl or GFR <40 ml / minute. Biochemical parameters (Glucose, AST, ALT, urea, creatinine, lipid levels, hemogram, calcium, phosphorus, parathormone) and the levels of serum Elabela were evaluated and compared. RESULTS: There was no significant difference in age (0.270) and gender (0.951) between groups. The median Elabela levels of the three groups were 40.3 ng/mL (22.5-54.6), 5.1 ng/mL (3.7-8.3), 9.2 ng/mL (6.1-23.1), respectively with a significant difference (p<0.001). CONCLUSION: The plasma levels of Elabela were lower in the case of hypertension, independent of nephropathy. However, this decrease is not specific for nephropathy and may be due to other accompanying chronic diseases. Key Words: Hypertension, Hypertensive nephropathy, Elabela.