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1.
Pest Manag Sci ; 77(1): 300-312, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32696609

RESUMO

BACKGROUND: The sweet potato weevil, Cylas formicarius, is the most serious pest of sweet potato worldwide. The molecular mechanism of sex pheromone recognition in C. formicarius has not been reported. Odorant-binding proteins (OBPs) play a critical role in selectively binding and transporting pheromones or other odors to the surface of olfactory receptor neurons through the aqueous sensillar lymph, therefore the function of sweet potato OBPs is worth studying. RESULTS: Herein, the CforOBP1-3 genes encoding three classical OBPs were cloned in C. formicarius by reverse transcription-polymerase chain reaction. Phylogenetic analysis showed that CforOBP1-3 were homologous genes, but the relationship between CforOBP2 and CforOBP3 was closest among the three genes. In addition, real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assays demonstrated that the expression of CforOBP1 was higher in the antennae and legs of female and male insects, while CforOBP2 and CforOBP3 were mainly expressed in the antennae of male insects. The fluorescent competitive binding assay results indicated that CforOBP1-3 had strong binding affinities to sex pheromones and other tested ligands. Finally, the mRNA expression of CforOBP1-3 was successfully inhibited by RNA interference, and in vivo behavioral experiments showed that CforOBP1-3-deficient C. formicarius was partly anosmic and lost some of its ability to locate sex pheromones and host plant volatiles. CONCLUSION: These results suggested that CforOBP1 was shown to be involved in the process of weevils feeding and finding sweet potato, and CforOBP2-3 were mainly involved in the mating behavior of adult male weevils.


Assuntos
Ipomoea batatas , Receptores Odorantes , Atrativos Sexuais , Gorgulhos , Animais , Proteínas de Transporte , Feminino , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Masculino , Odorantes , Percepção , Feromônios , Filogenia , Receptores Odorantes/genética , Receptores Odorantes/metabolismo , Gorgulhos/metabolismo
2.
Life Sci ; 264: 118632, 2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-33115605

RESUMO

Renal cell carcinoma (RCC) refers to renal-epithelial cancer, which represents over 90% of kidney cancer and is a cause for cancer related deaths in the world. Studies suggested somatic VHL mutations to be the cause for the occurrence of cancer, but with the time, more latest genomic and biological studies have detected variation in epigenetic regulatory genes and showed significant heterogeneity of the intratumor that may lead to strategies of diagnostic, predictive, and therapeutic importance. Immune dysfunction is responsible for almost all types of renal cancer, and angiogenesis and immunosuppression function together in the tumor microenvironment of renal cell carcinoma (RCC). Over the past few years, advancement in the management of the RCC has finally revolutionized with the arrival of the entrapped immune inhibitors which particularly concentrated on the receptor (programmed cell death-1) and focus on the new generation receptor i.e. TKRI (tyrosine-kinase receptor inhibitors). The present review deals with the comprehensive review of RCC and emphasizes on its types, pathogenesis and advancement in these diseases. This review also overviews the role of innate and adaptive immune response-related mechanism, the function of cancer stem cell in this diseases, therapeutic targeted drugs and hormonal signaling pathways as an emerging strategy in the management of the renal cancer.


Assuntos
Carcinoma de Células Renais/diagnóstico , Neoplasias Renais/diagnóstico , Mutação , Animais , Carcinoma de Células Renais/patologia , Carcinoma de Células Renais/terapia , Proteínas de Transporte/metabolismo , Progressão da Doença , Epigênese Genética , Hormônios/metabolismo , Humanos , Sistema Imunitário , Imunossupressão , Neoplasias Renais/patologia , Neoplasias Renais/terapia , Camundongos , Microcirculação , Terapia de Alvo Molecular , Células-Tronco Neoplásicas/metabolismo , Neovascularização Patológica/metabolismo , Fatores de Risco , Transdução de Sinais , Serina-Treonina Quinases TOR/metabolismo , Microambiente Tumoral , Proteínas Supressoras de Tumor/metabolismo , Ubiquitina Tiolesterase/metabolismo
3.
Antiviral Res ; 185: 104996, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33309540

RESUMO

Middle East Respiratory Syndrome (MERS) is a respiratory disease caused by a coronavirus (MERS-CoV). Since its emergence in 2012, nosocomial amplifications have led to its high epidemic potential and mortality rate of 34.5%. To date, there is an unmet need for vaccines and specific therapeutics for this disease. Available treatments are either supportive medications in use for other diseases or those lacking specificity requiring higher doses. The viral infection mode is initiated by the attachment of the viral spike glycoprotein to the human Dipeptidyl Peptidase IV (DPP4). Our attempts to screen antivirals against MERS led us to identify montelukast sodium hydrate (MSH), an FDA-approved anti-asthma drug, as an agent attenuating MERS-CoV infection. We showed that MSH directly binds to MERS-CoV-Receptor-Binding Domain (RBD) and inhibits its molecular interaction with DPP4 in a dose-dependent manner. Our cell-based inhibition assays using MERS pseudovirions demonstrated that viral infection was significantly inhibited by MSH and was further validated using infectious MERS-CoV culture. Thus, we propose MSH as a potential candidate for therapeutic developments against MERS-CoV infections.


Assuntos
Acetatos/farmacologia , Antivirais/farmacologia , Ciclopropanos/farmacologia , Coronavírus da Síndrome Respiratória do Oriente Médio/efeitos dos fármacos , Quinolinas/farmacologia , Sulfetos/farmacologia , Animais , Antiasmáticos/farmacologia , Proteínas de Transporte/efeitos dos fármacos , Chlorocebus aethiops , Infecções por Coronavirus/tratamento farmacológico , Indutores do Citocromo P-450 CYP1A2/farmacologia , Dipeptidil Peptidase 4/genética , Dipeptidil Peptidase 4/metabolismo , Reposicionamento de Medicamentos , Células HEK293 , Humanos , Antagonistas de Leucotrienos/farmacologia , Receptores Virais/genética , Receptores Virais/metabolismo , Glicoproteína da Espícula de Coronavírus/metabolismo , Células Vero , Internalização do Vírus/efeitos dos fármacos
4.
Life Sci ; 265: 118796, 2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-33220292

RESUMO

AIMS: In tumor cells, shikonin treatment has been reported to inhibit glycolysis by suppressing the activity of pyruvate kinase M2 (PKM2) and to induce apoptosis by increasing reactive oxygen species (ROS) production. However, hepatocellular carcinoma (HCC) shows variable sensitivity to shikonin treatment, and the mechanism for these differences remains unclear. We evaluated the effects of shikonin on metabolic and oxidative pathways in sensitive and refractory HCC cell lines to identify mechanisms of differential sensitivity. MAIN METHODS: Cell viability and apoptosis were evaluated by MTT assay, PI/Annexin V and JC-1 staining. Mitochondrial function was further evaluated by measurements of ROS and mitochondrial mass. Oxygen consumption rates, NAD+/NADH, ATP and lactate were measured as indicators of energy metabolism and glycolysis. Protein expression associated with glycolysis and apoptosis was evaluated by western blotting, RT-qPCR and immunofluorescence staining. KEY FINDINGS: The sensitivity to shikonin treatment was significantly higher for HepG2 cells than for HCCLM3 cells, with less dramatic effects in HCCLM3 cells on apoptosis, ROS, and oxidative phosphorylation. Shikonin up-regulated mitochondrial biogenesis to increase mitochondrial oxidative phosphorylation in HepG2 cells, but displayed the opposite trend in HCCLM3 cells. Mechanistically, shikonin promoted nuclear expression of PKM2 and HIF1α in HCCLM3 cells, with upregulation of glycolysis-related gene transcription and glycolysis. SIGNIFICANCE: These results suggest that PKM2 rewires glucose metabolism, which explains the differential sensitivity to shikonin-induced apoptosis in HCC cells. Our findings elucidate mechanisms for differential responses to shikonin, provide potential biomarkers, and indicate a theoretical basis for targeting glycolytic enzymes in refractory HCC.


Assuntos
Carcinoma Hepatocelular/metabolismo , Proteínas de Transporte/biossíntese , Glucose/metabolismo , Subunidade alfa do Fator 1 Induzível por Hipóxia/biossíntese , Neoplasias Hepáticas/metabolismo , Proteínas de Membrana/biossíntese , Naftoquinonas/farmacologia , Hormônios Tireóideos/biossíntese , Anti-Inflamatórios não Esteroides/farmacologia , Anti-Inflamatórios não Esteroides/uso terapêutico , Apoptose/efeitos dos fármacos , Apoptose/fisiologia , Carcinoma Hepatocelular/tratamento farmacológico , Relação Dose-Resposta a Droga , Glicólise/efeitos dos fármacos , Glicólise/fisiologia , Células Hep G2 , Humanos , Neoplasias Hepáticas/tratamento farmacológico , Naftoquinonas/uso terapêutico
5.
Life Sci ; 265: 118805, 2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-33245963

RESUMO

AIMS: To investigate the molecular function and mechanisms of JHDM1D antisense 1 (JHDM1D-AS1) during gastric cancer (GC) progression. MATERIALS AND METHODS: The qPCR assay was used to detect the JHDM1D-AS1 and miR-450a-2-3p expression levels in GC tissues and cell lines. Bioinformatics analysis was used for exploring the lncRNA-microRNA-mRNA interaction network. We performed dual-luciferase reporter assay and qPCR assay in order to validate the direct interactions. We explored the JHDM1D-AS1 and miR-450a-2-3p on GC progression by using JHDM1D-AS1 siRNA and miR-450a-2-3p inhibitor; in vitro CCK-8 assay, colony formation assay, and invasion assay were conducted. Further, in vivo animal experiments were performed, and the expression levels of miR-450a-2-3p and PRAF2 in the tumor tissues were detected using qPCR and western blot analysis. KEY FINDINGS: The expression levels of JHDM1D-AS1 and miR-450a-2-3p in GC tissues and cell lines were higher and lower as compared to those in the corresponding normal controls, respectively. Moreover, high levels of JHDM1D-AS1 were closely related with metastasis and the GC TNM stage. Functionally, JHDM1D-AS1 depletion caused an obvious reduction in cell proliferation and invasion both in vitro and in vivo, while the addition of miR-450a-2-3p inhibitor could nullify these effects. Mechanically, JHDM1D-AS1 promoted GC progression via the sponging of miR-450a-2-3p in order to increase PRAF2 expression. SIGNIFICANCE: The present results showed that the increased expression of JHDM1D-AS1 was closely associated with tumor progression of GC. JHDM1D-AS1/miR-450a-2-3p/PRAF2 axis may be a promising target for GC treatment.


Assuntos
Proteínas de Transporte/biossíntese , Progressão da Doença , Histona Desmetilases com o Domínio Jumonji/biossíntese , Proteínas de Membrana/biossíntese , MicroRNAs/biossíntese , Neoplasias Gástricas/metabolismo , Idoso , Animais , Proteínas de Transporte/genética , Linhagem Celular Tumoral , Feminino , Humanos , Histona Desmetilases com o Domínio Jumonji/genética , Masculino , Proteínas de Membrana/genética , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , MicroRNAs/genética , Pessoa de Meia-Idade , Invasividade Neoplásica/genética , Invasividade Neoplásica/patologia , RNA Antissenso/biossíntese , RNA Antissenso/genética , Transdução de Sinais/fisiologia , Neoplasias Gástricas/genética , Neoplasias Gástricas/patologia
7.
Gene ; 765: 145074, 2021 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-32891772

RESUMO

BARMR1/FAM92A1 encodes a novel BAR domain protein, and is widely expressed during embryonic development and highly expressed in tumor cells. Mutation or deletion of BARMR1/FAM92A1 caused developmental disorder and the BARMR1/FAM92A1 overexpression in tumor cells is associated with poor prognosis. The subcellular location of BARMR1/FAM92A1 determined its biological functions by interacting with different proteins. When colocalized and interacted with CBY at the centrioles/basal bodies of primary cilia, BARMR1/FAM92A1 facilitate ciliogenesis, whilst binded to GAL1 in the nuclei, it promotes cell proliferation, migration, and malignancy of tumor cells.


Assuntos
Proteínas/genética , Proteínas/metabolismo , Proteínas/fisiologia , Proteínas de Transporte/genética , Movimento Celular/genética , Proliferação de Células/genética , Centríolos/metabolismo , Cílios/genética , Galactoquinase/metabolismo , Humanos , Neoplasias/genética , Neoplasias/metabolismo , Proteínas Nucleares/genética
8.
Chemosphere ; 263: 128110, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33297103

RESUMO

Fish consumption is one of the major ways through which humans receive exposure to mercury (Hg). The existing forms of Hg in food, particularly Hg bound to proteins, may affect the absorption of Hg by humans and subsequently its potentially toxic effects. However, the knowledge regarding Hg-binding proteins in edible fish muscle is scarce. In the present study, salmon and tuna fish muscles, collected from seven different regions and countries, were analyzed using metallomics- and proteomics-based techniques. The concentration of Hg in sashimi samples ranged from 4.4 to 317.4 ng/g. Size exclusion chromatography (SEC) coupled with inductively coupled plasma mass spectrometer (ICP-MS) showed that beta-actin was a novel Hg-binding protein from the fish muscles, and this protein could also bind bismuth (Bi), silver (Ag), and copper (Cu). Hg bound to beta-actin accounted for approximately 30.2-37.6% of the total Hg in the tuna muscles and was significantly correlated to total Hg in the fish muscles (r = 0.98, p < 0.01) and in the fraction of soluble proteins (r = 0.94, p < 0.01). These findings suggest that proteins act as the main Hg accumulation sites in edible fish; thus, increasing human exposure to Hg following gastrointestinal digestion.


Assuntos
Mercúrio , Atum , Animais , Proteínas de Transporte , Contaminação de Alimentos/análise , Humanos , Mercúrio/análise , Salmão , Alimentos Marinhos/análise
9.
Life Sci ; 264: 118728, 2021 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-33160992

RESUMO

AIMS: Diabetic nephropathy (DN) is the most frequent complication of diabetes and causes millions of deaths each year. Finding novel therapy to DN is urgent, which requires a good understanding of the pathogenesis. Aims are to investigate the molecular mechanisms of DN by focusing on ANRIL/miR-497/TXNIP axis. MAIN METHODS: Kidney tissues were collected from diagnosed DN patients. High glucose (HG) treatment of human renal tubular epithelial cell cells (HK-2) was used as the cell model of DN. qRT-PCR and Western blotting were performed to measure levels of ANRIL, miR-497, TXNIP, IL-1ß, IL-18, caspase-1, and NLRP3. LDH leakage and cell viability were determined with commercial LDH activity kit and MTT assay. ELISA was employed to examine secreted IL-1ß and IL-18 levels. Flow cytometry was used to examine caspase-1 activity. Dual luciferase assay was performed to validate interactions of ANRIL/miR-497 and miR-497/TXNIP. KEY FINDINGS: ANRIL and TXNIP were elevated in DN kidney tissues and HG-treated HK-2 cells while miR-497 was reduced. ANRIL bound miR-497 while miR-497 directly targeted TXNIP. Knockdown of ANRIL suppressed HG-induced LDH leakage, TXNIP/NLRP3/caspase-1 activation, and increases of IL-1ß and IL-18 secreted levels. miR-497 knockdown or TXNIP overexpression reversed the effects of ANRIL knockdown on LDH leakage and pyroptosis-related signaling. miR-497 mimics inhibited caspase-1-dependent pyroptosis while co-overexpression of TXNIP blocked its effects in HG-treated HK-2 cells. SIGNIFICANCE: ANRIL promotes pyroptosis and kidney injury in DN via acting as miR-497 sponge to disinhibit TXNIP expression. These results shed light on the mechanisms of DN and provide targets for therapy development.


Assuntos
Proteínas de Transporte/metabolismo , Nefropatias Diabéticas/genética , MicroRNAs/metabolismo , Piroptose/genética , RNA Longo não Codificante/genética , Sequência de Bases , Proteínas de Transporte/genética , Caspase 1/metabolismo , Linhagem Celular , Técnicas de Silenciamento de Genes , Glucose/toxicidade , Humanos , MicroRNAs/genética , Modelos Biológicos , Piroptose/efeitos dos fármacos , RNA Longo não Codificante/metabolismo , Transdução de Sinais/efeitos dos fármacos
10.
Zhonghua Nan Ke Xue ; 26(3): 258-264, 2020 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-33346967

RESUMO

Objective: To investigate the effects of Xiongcan Yishen Prescription (XYP) on the expressions of cholesterol transport proteins, steroidogenic enzymes and steroidogenic factor-1 (SF-1) in the Leydig cells of the rats with late-onset hypogonadism (LOH). METHODS: Twenty-five 18-month-old male SD rats were randomly divided into five groups of equal number, LOH model control, testosterone propionate (TP) and low-, medium- and high-dose XYP, and another 5 two-month-old male SD rats included as normal controls. After modeling, the animals in the TP group were treated by intramuscular injection of TP at 5.21 mg/kg qd alt, those in the low-, medium- and high-dose XYP groups intragastrically with XYP at 10.4, 20.8 and 41.6 g/kg qd alt respectively, and those in the LOH model and normal control groups with saline, all for 28 successive days. Then, all the rats were sacrificed for determination of the expressions of the cholesterol transport proteins StAR and TSPO, steroidogenic enzymes CYP11A1, HSD3B7 and HSD17B4, and SF-1 in the Leydig cells by Western blot. RESULTS: The expressions of StAR, TSPO, CYP11A1, HSD3B7, HSD17B4 and SF-1 in the Leydig cells were significantly decreased in the LOH model controls compared with those in the normal controls (P< 0.05), but remarkably increased in the low-, medium- and high-dose XYP groups in comparison with those in the LOH model control group (P< 0.05). CONCLUSIONS: Xiongcan Yishen Prescription can up-regulate the expressions of the cholesterol transport proteins StAR and TSPO, steroidogenic enzymes CYP11A1, HSD3B7 and HSD17B4, and SF-1 in the rat Leydig cells, which might be one of the possible mechanisms of the prescription in the treatment of LOH.


Assuntos
Colesterol/metabolismo , Medicamentos de Ervas Chinesas/uso terapêutico , Hidroxiesteroide Desidrogenases/metabolismo , Hipogonadismo , Células Intersticiais do Testículo/efeitos dos fármacos , Animais , Transporte Biológico , Proteínas de Transporte , Hipogonadismo/tratamento farmacológico , Células Intersticiais do Testículo/metabolismo , Masculino , Fosfoproteínas/metabolismo , Ratos , Ratos Sprague-Dawley , Receptores de Esteroides/metabolismo , Testosterona
11.
Nat Commun ; 11(1): 6114, 2020 11 30.
Artigo em Inglês | MEDLINE | ID: mdl-33257653

RESUMO

DNA methylation differences in Alzheimer's disease (AD) have been reported. Here, we conducted a meta-analysis of more than 1000 prefrontal cortex brain samples to prioritize the most consistent methylation differences in multiple cohorts. Using a uniform analysis pipeline, we identified 3751 CpGs and 119 differentially methylated regions (DMRs) significantly associated with Braak stage. Our analysis identified differentially methylated genes such as MAMSTR, AGAP2, and AZU1. The most significant DMR identified is located on the MAMSTR gene, which encodes a cofactor that stimulates MEF2C. Notably, MEF2C cooperates with another transcription factor, PU.1, a central hub in the AD gene network. Our enrichment analysis highlighted the potential roles of the immune system and polycomb repressive complex 2 in pathological AD. These results may help facilitate future mechanistic and biomarker discovery studies in AD.


Assuntos
Doença de Alzheimer/genética , Doença de Alzheimer/metabolismo , Metilação de DNA/fisiologia , Epigênese Genética/genética , Imunidade/genética , Córtex Pré-Frontal/fisiologia , Idoso , Idoso de 80 Anos ou mais , Doença de Alzheimer/imunologia , Encéfalo , Proteínas de Transporte/genética , Epigênese Genética/fisiologia , Feminino , Proteínas de Ligação ao GTP/genética , Proteínas Ativadoras de GTPase/genética , Redes Reguladoras de Genes , Estudo de Associação Genômica Ampla , Humanos , Imunidade/fisiologia , Fatores de Transcrição MEF2/genética , Fatores de Transcrição/genética , Proteínas Supressoras de Tumor/genética
12.
Zhonghua Yi Xue Za Zhi ; 100(48): 3863-3869, 2020 Dec 29.
Artigo em Chinês | MEDLINE | ID: mdl-33371632

RESUMO

Objective: To investigate the role and regulation mechanism of X box binding protein 1 (XBP1) for hypoxia/reoxygenation(H/R) injury in mouse renal tubular epithelial cells (TCMK-1) through thioredoxin interacting protein (TXNIP)-nucleotide-binding domain (NOD)-like receptor protein (TXNIP-NLRP3) signaling pathway. Methods: The cells were divided into 4 groups: si-NC group transfected with negative control siRNA (si-NC), si-XBP1 group transfected with siRNA targeting XBP1 (si-XBP1), si-NC+H/R group transfected with si-NC and exposed to H/R, and si-XBP1+H/R group transfected with si-XBP1 and exposed to H/R. The Annexin Ⅴ/PI double-staining method was used to detect cell apoptosis; The mitochondrial membrane potential (MMP) was determined by using JC-1 dye; The mitochondrial reactive oxygen species (mROS) was assessed by using MitoSOX™ dye. The interference efficiency of XBP1 was tested by Western blotting and quantitative real-time polymerase chain reaction. The expression levels of TXNIP, NLRP3 and IL-1ß protein were detected by Western blotting. The colocalization of mitochondria and TXNIP was detected by double-labeling immunofluorescent staining. The intergroup difference was compared by using an independent samples t-test. Results: Compared with the si-NC group, more mROS, apoptosis and lower MMP were observed in si-NC+H/R group. Compared with the si-NC+H/R group, less apoptosis (12.08±0.51 vs 19.01±1.80, P<0.05), mROS (34.63±0.64 vs 48.17±1.84, P<0.01) and higher MMP (1.03±0.11 vs 0.45±0.08, P<0.05) were observed in si-XBP1+H/R group. Down-regulation of XBP1U (protein: 1.31±0.18 vs 0.23±0.02, P<0.01; mRNA: 1.12±0.07 vs 0.38±0.01, P<0.001) and XBP1S (protein: 1.13±0.17 vs 0.28±0.07, P<0.01; mRNA: 8.39±0.63 vs 2.45±0.22, P<0.001) inhibited expression of TXNIP (0.15±0.02 vs 0.04±0.01, P<0.01), NLRP3 (1.13±0.12 vs 0.51±0.12, P<0.05) and IL-1ß (1.02±0.04 vs 0.19±0.06, P<0.001) during H/R. Meanwhile, TXNIP exhibited significantly much less colocalization with mitochondria in the si-XBP1+H/R group. Conclusion: Supression of XBP1 expression can effectively alleviate H/R-induced TCMK-1 cells injury, whose mechanism may be inhibition of TXNIP-induced NLRP3 inflammasome activation.


Assuntos
Inflamassomos , Proteína 3 que Contém Domínio de Pirina da Família NLR , Animais , Proteínas de Transporte , Células Epiteliais/metabolismo , Hipóxia , Inflamassomos/metabolismo , Camundongos , Proteína 3 que Contém Domínio de Pirina da Família NLR/genética , Proteína 3 que Contém Domínio de Pirina da Família NLR/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais , Tiorredoxinas/metabolismo , Proteína 1 de Ligação a X-Box/genética
13.
Environ Pollut ; 267: 115568, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33254717

RESUMO

Previous studies have associated the risk of autism spectrum disorder (ASD) with increased exposures to metals and metalloids such as arsenic. In this study, we used an animal-to-human translational strategy to identify key molecular changes that potentially mediated the effects of arsenic exposures on ASD development. In a previously established rat model, we have induced autistic behaviors in rat pups with gestational arsenic exposures (10 and 45 µg/L As2O3 in drinking water). Neuronal apoptosis and the associated epigenetic dysregulations in frontal cortex were assayed to screen potential mediating pathways, which were subsequently validated with qPCR, western blotting, and immunohistochemistry analyses. Furthermore, the identified pathway, along with serum levels of 26 elements including arsenic, were characterized in a case-control study with 21 ASD children and 21 age-matched healthy controls. In animals, we found that arsenic exposures caused difficulties of social interaction and increased stereotypic behaviors in a dose-dependent manner, accompanied by increased neuronal apoptosis and upregulation of Hipk2-p53 pathway in the frontal cortex. In humans, we found that serum levels of Hipk2 and p53 were 24.7 (95%CI: 8.5 to 43.4) % and 23.7 (95%CI: 10.5 to 38.5) % higher in ASD children than in healthy controls. ASD children had significantly higher serum levels of 15 elements, among which arsenic, silicon, strontium, and vanadium were positively associated with both Hipk2 and p53. Results from both the rat arsenic exposure and human case-control studies suggest a likely role of Hipk2-p53 pathway in ASD development induced by exposures to environmental pollutants such as arsenic.


Assuntos
Arsênico , Transtorno do Espectro Autista , Transtorno Autístico , Animais , Arsênico/toxicidade , Transtorno do Espectro Autista/induzido quimicamente , Transtorno Autístico/induzido quimicamente , Proteínas de Transporte , Estudos de Casos e Controles , Criança , Humanos , Proteínas Serina-Treonina Quinases , Ratos , Proteína Supressora de Tumor p53/genética
14.
Fa Yi Xue Za Zhi ; 36(5): 672-676, 2020 Oct.
Artigo em Chinês | MEDLINE | ID: mdl-33295169

RESUMO

Abstract: Objective To explore the correlation between 4 tag single nucleotide polymorphisms (Tag SNP) sites (rs7721799, rs32897, rs7718461, rs10062367) of corticotropin releasing hormone binding protein (CRHBP) and schizophrenia and aggressive behavior in the Yunnan Han population. Methods Case-control correlation analysis was used to establish a complex amplification system. Improved multiplex ligase detection reaction (iMLDR) technology was used to detect the genotypes of 4 SNP sites of CRHBP gene of 163 Han schizophrenic patients (including 81 patients with aggressive behavior, 82 patients without aggressive behavior) and 345 healthy Han individuals, which were analyzed statistically by SPSS 19.0, Haploview 4.2 and PHASE 2.1 software. Results There was no correlation between the 3 SNP sites of CRHBP gene and the onset of schizophrenia except for the rs7718461 site (P>0.05). The relative risk of aggressive behavior of patients carrying GG or GA genotype at rs7718461 site were 4.903 times higher than those carrying AA genotype (P<0.05). Conclusion The CRHBP gene may not be associated with the occurrence of schizophrenia in Yunnan Han population, but AA genotype of rs7718461 may reduce the risk of aggressive behavior in schizophrenia patients.


Assuntos
Proteínas de Transporte , Esquizofrenia , Grupo com Ancestrais do Continente Asiático/genética , Proteínas de Transporte/genética , China , Frequência do Gene , Predisposição Genética para Doença , Genótipo , Humanos , Polimorfismo de Nucleotídeo Único , Esquizofrenia/genética
15.
PLoS One ; 15(11): e0241112, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33232330

RESUMO

Natural α-tocopherol (α-TCP), but not tocotrienol, is preferentially retained in the human body. α-Tocopherol transfer protein (α-TTP) is responsible for binding α-TCP for cellular uptake and has high affinity and specificity for α-TCP but not α-tocotrienol. The purpose of this study was to examine the modification of α-TTP together with other related vitamin E-binding genes (i.e., TTPA, SEC14L2, and PI-TPNA) in regulating vitamin E uptake in neuronal cells at rest and under oxidative stress. Oxidative stress was induced with H2O2 for an hour which was followed by supplementation with different ratios of α-TCP and tocotrienol-rich fraction (TRF) for four hours. The cellular levels of vitamin E were quantified to determine bioavailability at cellular levels. The expression levels of TTPA, SEC14L2, and PI-TPNA genes in 0% α-TCP were found to be positively correlated with the levels of vitamin E in resting neuronal cells. In addition, the regulation of all the above-mentioned genes affect the distribution of vitamin E in the neuronal cells. It was observed that, increased levels of α-TCP secretion occur under oxidative stress. Thus, our results showed that in conclusion vitamin E-binding proteins may be modified in the absence of α-TCP to produce tocotrienols (TCT), as a source of vitamin E. The current study suggests that the expression levels of vitamin E transport proteins may influence the cellular concentrations of vitamin E levels in the neuronal cells.


Assuntos
Peróxido de Hidrogênio/farmacologia , Neurônios/efeitos dos fármacos , Neurônios/metabolismo , Tocotrienóis/farmacologia , Vitamina E/metabolismo , alfa-Tocoferol/farmacologia , Antioxidantes/metabolismo , Disponibilidade Biológica , Transporte Biológico/fisiologia , Proteínas de Transporte/metabolismo , Linhagem Celular Tumoral , Suplementos Nutricionais , Humanos , Fármacos Neuroprotetores/farmacologia , Estresse Oxidativo/efeitos dos fármacos
16.
Medicine (Baltimore) ; 99(45): e23033, 2020 Nov 06.
Artigo em Inglês | MEDLINE | ID: mdl-33157955

RESUMO

INTRODUCTION: Microdeletion syndromes occur from deletion of 5Mb of a chromosome in approximately 5% of patients with unexplained intellectual disability. Interstitial microdeletions at bands 1p33 and 1p32.2 of the short arm of chromosome 1 are rare and have not been previously reported in relation to disease. PATIENT CONCERNS: We present a case of a 39-month boy with Pierre Robin sequence, development delay/intellectual disability, growth retardation, short stature, leukoencephalopathy, craniofacial dysplasia, and speech delay. The child was referred to the Child health care department in October 2014 for his delayed language development and aggravated aggression. DIAGNOSIS: Molecular diagnostic testing with G-band karyotyping was normal but clinical microarray analysis detected a 10 Mb microdeletion at 1p33p32.2. INTERVENTIONS: The patient received rehabilitation. OUTCOMES: Three candidate genes were pinpointed to the deleted area, including ORC1, SCP2, and DAB1. Phenotype-genotype analysis suggested that these three genes are likely to be responsible for the main phenotypes observed in the patient, such as microcephaly, growth retardation, short stature, leukoencephalopathy, and development delay/intellectual disability. CONCLUSIONS: The spectrum of phenotypes this case presented with are likely to be caused by 1p33p32.2 deletion which could represent a new microdeletion syndrome.


Assuntos
Cariotipagem/métodos , Análise em Microsséries/métodos , Síndrome de Pierre Robin/diagnóstico , Síndrome de Pierre Robin/genética , Anormalidades Múltiplas/diagnóstico , Anormalidades Múltiplas/genética , Proteínas Adaptadoras de Transdução de Sinal/genética , Adolescente , Adulto , Proteínas de Transporte/genética , Criança , Pré-Escolar , Deleção Cromossômica , Anormalidades Craniofaciais/diagnóstico , Anormalidades Craniofaciais/genética , Nanismo/diagnóstico , Nanismo/genética , Feminino , Humanos , Lactente , Deficiência Intelectual/diagnóstico , Deficiência Intelectual/genética , Transtornos do Desenvolvimento da Linguagem/diagnóstico , Transtornos do Desenvolvimento da Linguagem/genética , Leucoencefalopatias/diagnóstico , Leucoencefalopatias/genética , Masculino , Microcefalia/diagnóstico , Microcefalia/genética , Proteínas do Tecido Nervoso/genética , Complexo de Reconhecimento de Origem/genética , Fenótipo , Síndrome de Pierre Robin/reabilitação
17.
Proc Natl Acad Sci U S A ; 117(47): 29712-29719, 2020 11 24.
Artigo em Inglês | MEDLINE | ID: mdl-33168715

RESUMO

The ammonium transporter (AMT)/methylammonium permease (MEP)/Rhesus glycoprotein (Rh) family of ammonia (NH3/NH4 +) transporters has been identified in organisms from all domains of life. In animals, fundamental roles for AMT and Rh proteins in the specific transport of ammonia across biological membranes to mitigate ammonia toxicity and aid in osmoregulation, acid-base balance, and excretion have been well documented. Here, we observed enriched Amt (AeAmt1) mRNA levels within reproductive organs of the arboviral vector mosquito, Aedes aegypti, prompting us to explore the role of AMTs in reproduction. We show that AeAmt1 is localized to sperm flagella during all stages of spermiogenesis and spermatogenesis in male testes. AeAmt1 expression in sperm flagella persists in spermatozoa that navigate the female reproductive tract following insemination and are stored within the spermathecae, as well as throughout sperm migration along the spermathecal ducts during ovulation to fertilize the descending egg. We demonstrate that RNA interference (RNAi)-mediated AeAmt1 protein knockdown leads to significant reductions (∼40%) of spermatozoa stored in seminal vesicles of males, resulting in decreased egg viability when these males inseminate nonmated females. We suggest that AeAmt1 function in spermatozoa is to protect against ammonia toxicity based on our observations of high NH4 + levels in the densely packed spermathecae of mated females. The presence of AMT proteins, in addition to Rh proteins, across insect taxa may indicate a conserved function for AMTs in sperm viability and reproduction in general.


Assuntos
Aedes/metabolismo , Compostos de Amônio/metabolismo , Proteínas de Transporte/metabolismo , Proteínas de Insetos/metabolismo , Mosquitos Vetores/metabolismo , Amônia/metabolismo , Animais , Vetores de Doenças , Fertilidade/fisiologia , Fertilização/fisiologia , Masculino , RNA Mensageiro/metabolismo , Reprodução/fisiologia , Espermatogênese/fisiologia , Espermatozoides/metabolismo , Testículo/metabolismo
18.
Seizure ; 83: 145-153, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-33152654

RESUMO

PURPOSE: To elucidate the presenting symptoms of Lafora Disease (LD) to differentiate it from Juvenile Myoclonic Epilepsy (JME). METHODS: We collected and evaluated the early electroclinical data of 5 unrelated Apulian (Southern Italy) LD families, 30 LD patients selected from the literature, and 30 Apulian JME patients. RESULTS: The Apulian LD patients presented with generalised tonic-clonic and focal visual seizures, followed by myoclonic seizures and action-postural myoclonus. In these patients, EEG background slowing and occipital epileptiform abnormalities were significantly more evident than in the other groups. Genetic analysis revealed the presence of mutations in the EPM2A gene in 4 families, and in the NHLRC1 gene in the remaining family. In detail, we identified 2 different point mutations in EPM2A and only 1 in NHLRC1, and expanded the molecular spectrum of the EPM2A gene mutations reporting for the first time a patient carrier of the c.243_246del genetic variant. In the previously reported LD cases, generalised tonic-clonic and focal visual seizures and myoclonus were the most frequent symptoms, as confirmed by the first EEGs showing occipital or diffuse epileptiform abnormalities with photosensitivity in the background activity slowing. In the Apulian JME patients, myoclonus appeared earlier, usually at awakening, with diffuse epileptiform abnormalities during sleep and photosensitivity in the normal background activity. The diagnosis of JME was established much earlier than the LD one. During evolution, unlike JME patients, LD patients showed a significant resistance to drugs. CONCLUSIONS: Tonic-clonic and focal visual seizures followed by myoclonic seizures and action-postural myoclonus together with EEG background slowing with diffuse and occipital epileptiform abnormalities suggest a diagnosis of LD. An early molecular confirmation allows a better diagnosis, counselling and management of affected patients and their families, and it may be useful to improve the patients' quality of life using, when possible, emerging personalized treatments that may slow the evolution of the disease.


Assuntos
Doença de Lafora/genética , Doença de Lafora/fisiopatologia , Mutação/genética , Epilepsia Mioclônica Juvenil/genética , Convulsões/genética , Adolescente , Adulto , Proteínas de Transporte/genética , Criança , Feminino , Testes Genéticos , Humanos , Itália , Masculino , Proteínas Tirosina Fosfatases não Receptoras/genética , Qualidade de Vida , Adulto Jovem
19.
PLoS Biol ; 18(11): e3000904, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-33156822

RESUMO

There is a great need for antiviral drugs to treat enterovirus (EV) and rhinovirus (RV) infections, which can be severe and occasionally life-threatening. The conserved nonstructural protein 2C, which is an AAA+ ATPase, is a promising target for drug development. Here, we present a structure-activity relationship study of a previously identified compound that targets the 2C protein of EV-A71 and several EV-B species members, but not poliovirus (PV) (EV-C species). This compound is structurally related to the Food and Drug Administration (FDA)-approved drug fluoxetine-which also targets 2C-but has favorable chemical properties. We identified several compounds with increased antiviral potency and broadened activity. Four compounds showed broad-spectrum EV and RV activity and inhibited contemporary strains of emerging EVs of public health concern, including EV-A71, coxsackievirus (CV)-A24v, and EV-D68. Importantly, unlike (S)-fluoxetine, these compounds are no longer neuroactive. By raising resistant EV-A71, CV-B3, and EV-D68 variants against one of these inhibitors, we identified novel 2C resistance mutations. Reverse engineering of these mutations revealed a conserved mechanism of resistance development. Resistant viruses first acquired a mutation in, or adjacent to, the α2 helix of 2C. This mutation disrupted compound binding and provided drug resistance, but this was at the cost of viral fitness. Additional mutations at distantly localized 2C residues were then acquired to increase resistance and/or to compensate for the loss of fitness. Using computational methods to identify solvent accessible tunnels near the α2 helix in the EV-A71 and PV 2C crystal structures, a conserved binding pocket of the inhibitors is proposed.


Assuntos
Antivirais/farmacologia , Proteínas de Transporte/efeitos dos fármacos , Enterovirus/efeitos dos fármacos , Proteínas não Estruturais Virais/efeitos dos fármacos , Antígenos Virais , Proteínas de Transporte/metabolismo , Descoberta de Drogas/métodos , Enterovirus/patogenicidade , Infecções por Enterovirus/virologia , Fluoxetina/farmacologia , Células HeLa , Humanos , Relação Estrutura-Atividade , Proteínas não Estruturais Virais/metabolismo , Replicação Viral
20.
PLoS Genet ; 16(11): e1009187, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-33137092

RESUMO

Germline mutations in the folliculin (FLCN) tumor suppressor gene are linked to Birt-Hogg-Dubé (BHD) syndrome, a dominantly inherited genetic disease characterized by predisposition to fibrofolliculomas, lung cysts, and renal cancer. Most BHD-linked FLCN variants include large deletions and splice site aberrations predicted to cause loss of function. The mechanisms by which missense variants and short in-frame deletions in FLCN trigger disease are unknown. Here, we present an integrated computational and experimental study that reveals that the majority of such disease-causing FLCN variants cause loss of function due to proteasomal degradation of the encoded FLCN protein, rather than directly ablating FLCN function. Accordingly, several different single-site FLCN variants are present at strongly reduced levels in cells. In line with our finding that FLCN variants are protein quality control targets, several are also highly insoluble and fail to associate with the FLCN-binding partners FNIP1 and FNIP2. The lack of FLCN binding leads to rapid proteasomal degradation of FNIP1 and FNIP2. Half of the tested FLCN variants are mislocalized in cells, and one variant (ΔE510) forms perinuclear protein aggregates. A yeast-based stability screen revealed that the deubiquitylating enzyme Ubp15/USP7 and molecular chaperones regulate the turnover of the FLCN variants. Lowering the temperature led to a stabilization of two FLCN missense proteins, and for one (R362C), function was re-established at low temperature. In conclusion, we propose that most BHD-linked FLCN missense variants and small in-frame deletions operate by causing misfolding and degradation of the FLCN protein, and that stabilization and resulting restoration of function may hold therapeutic potential of certain disease-linked variants. Our computational saturation scan encompassing both missense variants and single site deletions in FLCN may allow classification of rare FLCN variants of uncertain clinical significance.


Assuntos
Síndrome de Birt-Hogg-Dubé/patologia , Complexo de Endopeptidases do Proteassoma/metabolismo , Proteínas Proto-Oncogênicas/metabolismo , Proteínas Supressoras de Tumor/metabolismo , Síndrome de Birt-Hogg-Dubé/genética , Proteínas de Transporte/metabolismo , Linhagem Celular Tumoral , Biologia Computacional , Mutação em Linhagem Germinativa , Humanos , Mutação com Perda de Função , Mutação de Sentido Incorreto , Agregados Proteicos , Ligação Proteica/genética , Dobramento de Proteína , Estabilidade Proteica , Proteólise , Proteínas Proto-Oncogênicas/genética , Saccharomyces cerevisiae , Proteínas Supressoras de Tumor/genética , Peptidase 7 Específica de Ubiquitina/metabolismo
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