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Background: The contribution of total fat mass and regional fat distribution to the risk of AF has rarely been studied. Methods: This prospective cohort study(N=494,063) evaluated the association of total fat mass measured by fat percentage (FP) and regional fat measured by arm fat percentage (AFP), trunk fat percentage (TFP), and leg fat percentage (LFP) with incident AF. A subgroup (N = 25,581) underwent MRI, which allowed us to further assess whether visceral adipose tissue (VAT) and abdominal subcutaneous adipose tissue (ASAT) of the trunk fat exert different effects on AF incidence. Results: Over, a median 12.9 ± 1.86 years of follow-up, 29,658 participants (cumulative rate: 6.0%) developed AF. Each 1-standard deviation (SD) increase in LFP was associated with a 16% lower risk of AF (HR: 0.84, 95% CI: 0.82, 0.85). The association between FP and AF was weaker than that between LFP and AF (HR: 0.90, 95% CI: 0.89, 0.92). AFP and TFP only had a marginal association with a lower incidence of AF. Both the VAT and ASAT showed a U-shaped relationship with incident AF. Conclusions: Fat mass, mainly leg fat mass, was associated with a lower risk of AF. ASAT did not exert protective effects.
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Fibrilación Atrial , Humanos , Fibrilación Atrial/epidemiología , Fibrilación Atrial/etiología , Estudios de Cohortes , Estudios Prospectivos , Factores de Riesgo , alfa-FetoproteínasRESUMEN
Soil salinization is one of the most serious abiotic stresses restricting plant growth. Buffalograss is a C4 perennial turfgrass and forage with an excellent resistance to harsh environments. To clarify the adaptative mechanisms of buffalograss in response to salinity, we investigated the effects of NaCl treatments on photosynthesis, water status and K+/Na+ homeostasis of this species, then analyzed the expression of key genes involved in these processes using the qRT-PCR method. The results showed that NaCl treatments up to 200 mM had no obvious effects on plant growth, photosynthesis and leaf hydrate status, and even substantially stimulated root activity. Furthermore, buffalograss could retain a large amount of Na+ in roots to restrict Na+ overaccumulation in shoots, and increase leaf K+ concentration to maintain a high K+/Na+ ratio under NaCl stresses. After 50 and 200 mM NaCl treatments, the expressions of several genes related to chlorophyll synthesis, photosynthetic electron transport and CO2 assimilation, as well as aquaporin genes (BdPIPs and BdTIPs) were upregulated. Notably, under NaCl treatments, the increased expression of BdSOS1, BdHKT1 and BdNHX1 in roots might have helped Na+ exclusion by root tips, retrieval from xylem sap and accumulation in root cells, respectively; the upregulation of BdHAK5 and BdSKOR in roots likely enhanced K+ uptake and long-distance transport from roots to shoots, respectively. This work finds that buffalograss possesses a strong ability to sustain high photosynthetic capacity, water balance and leaf K+/Na+ homeostasis under salt stress, and lays a foundation for elucidating the molecular mechanism underlying the salt tolerance of buffalograss.
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Hydrogen sulfide (H2 S) is an important gaseous signal molecule that regulates plant growth and stress resistance. However, research on the H2 S synthase (HSase) genes is still limited in the model legume plant Medicago truncatula Gaertn. In the present study, a total of 40 HSase family members were first identified and analyzed in the M. truncatula genome, and these genes distributed across eight chromosomes and were clustered into five groups (I-V) based on their conserved gene structures and protein motifs. Expression analysis revealed that the MtHSase genes were expressed in all the tested abiotic stresses, albeit with expression level differences. This study also showed that H2 S improves low temperature tolerance of alfalfa seedlings by regulating the antioxidant defense system and enhancing photosynthetic capacity. Thus, the study provides new insights into how the H2 S signal regulates tolerance to low-temperature stress and provides the basis for further gene function and detection.
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Sulfuro de Hidrógeno , Medicago truncatula , Medicago sativa/genética , Sulfuro de Hidrógeno/metabolismo , Proteínas de Plantas/metabolismo , Temperatura , Plantas/metabolismo , Estrés Fisiológico/genética , Regulación de la Expresión Génica de las Plantas , Medicago truncatula/genética , FilogeniaRESUMEN
OBJECTIVES: Exploring the role of OPN N-glycosylation in osteoblasts and osteoclasts. METHODS: Immunohistochemistry was used to detect the expression of OPN in mice with apical periodontitis. The asparagine at position 79 of the OPN protein was mutated to glutamine, and the above plasmids were transfected into osteoblasts and osteoclasts. The effect of OPN N-glycosylation on proliferation of osteoblasts and osteoclasts was detected by CCK8 assays. Western blotting was used to detect the expression of OPN N-glycosylation on osteoclasts and osteoblasts. Detection of N-glycosylation of OPN activated the NF-κB signaling pathway to regulate osteoblasts and osteoclasts. RESULTS: OPN increased the expression in a mice model of apical periodontitis. The expression curve of OPN resembled a reverse V shape. The OPN N-glycosylation site was identified as 79 by MS. N-glycosylation of OPN promoted the proliferation of osteoclasts. But the N79 glycosylation site of mutant OPN could not increase the proliferation of osteoblasts. OPN N-glycosylation modulated the expression of osteoclast- and osteoblast-associated factors through the NF-κB signaling pathway. N-glycosylation of OPN promoted nuclear translocation of NF-κB in osteoclasts and osteoblasts. CONCLUSIONS: The N-glycosylation site of OPN is 79. N-glycosylation of OPN played an important role in the biological function of OPN protein.
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FN-kappa B , Periodontitis Periapical , Ratones , Animales , FN-kappa B/metabolismo , Osteopontina/metabolismo , Glicosilación , Osteoclastos/metabolismo , Osteoblastos/metabolismo , Periodontitis Periapical/metabolismo , Diferenciación CelularRESUMEN
Soil salinity has become one of the major factors that threaten tall fescue growth and turf quality. Plants recruit diverse microorganisms in the rhizosphere to cope with salinity stress. In this study, 15 plant growth-promoting rhizobacteria (PGPR) were isolated from the salt-treated rhizosphere of tall fescue and were annotated to 10 genera, including Agrobacterium, Fictibacillus, Rhizobium, Bhargavaea, Microbacterium, Paenarthrobacter, Pseudarthrobacter, Bacillus, Halomonas, and Paracoccus. All strains could produce indole-3-acetic acid (IAA). Additionally, eight strains exhibited the ability to solubilize phosphate and potassium. Most strains could grow on the medium containing 600 mM NaCl, such as Bacillus zanthoxyli and Bacillus altitudinis. Furthermore, Bacillus zanthoxyli and Bacillus altitudinis were inoculated with tall fescue seeds and seedlings to determine their growth-promoting effect. The results showed that Bacillus altitudinis and mixed culture significantly increased the germination rate of tall fescue seeds. Bacillus zanthoxyli can significantly increase the tillers number and leaf width of seedlings under salt conditions. Through the synergistic effect of FaSOS1, FaHKT1, and FaHAK1 genes, Bacillus zanthoxyli helps to expel the excess Na+ from aboveground parts and absorb more K+ in roots to maintain ion homeostasis in tall fescue. Unexpectedly, we found that Bacillus altitudinis displayed an inapparent growth-promoting effect on seedlings under salt stress. Interestingly, the mixed culture of the two strains was also able to alleviate, to some extent, the effects of salt stress on tall fescue. This study provides a preliminary understanding of tall fescue rhizobacteria and highlights the role of Bacillus zanthoxyli in tall fescue growth and salt tolerance.
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Bacillus , Festuca , Lolium , Rizosfera , Estrés Salino , Desarrollo de la Planta , Plantones , Raíces de PlantasRESUMEN
Biochemistry, a complicated basic course in health sciences, plays a fundamental role in describing pathogenic mechanism of illness in molecular terms, and is required to be learned for all medical students. Due to various obstacles to biochemistry learning and teaching, there is a pressing issue of curriculum reform to arouse the student's enthusiasm in biochemistry learning. In this study, an integrated topic-based biochemistry training program (TBBTP) combining the traditional lectures, question-based learning and experimental practices, was introduced in biochemistry teaching. Its effectiveness was evaluated through examination and questionnaire analyses. Consequently, we found that TBBTP promoted the dissemination of biochemical knowledge via traditional lecture, designated learning issues and experimental practices, and acquisition of various skills through practical operation, presentation, and independent learning. It motivated students to study biochemistry with intense interest and enthusiasm. This study explored the feasibility of the topic-based biochemistry teaching as a supplement to biochemistry curriculum in medical education and as a mainstream pedagogy in biochemistry lab.
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Educación de Pregrado en Medicina , Estudiantes de Medicina , Humanos , Curriculum , Bioquímica/educación , Aprendizaje , EnseñanzaRESUMEN
Paclobutrazol (PBZ) is a plant-growth regulator (PGR) in the triazole family that enhances plant tolerance to environmental stresses. Low-light (LL) intensity is a critical factor adversely affecting the growth of tall fescue (Festuca arundinacea Schreb.). Therefore, in this study, tall fescue seedlings were treated with PBZ under control and LL conditions to investigate the effects of PBZ on enhancing LL stress resistance by regulating the growth, photosynthesis, oxidative defense, and hormone levels. Our results reveal that LL stress reduced the total biomass, chlorophyll (Chl) content, photosynthetic capacity, and photochemical efficiency of photosystem II (PSII) but increased the membrane lipid peroxidation level and reactive oxygen species (ROS) generation. However, the application of PBZ increased the photosynthetic pigment contents, net photosynthetic rate (Pn), maximum quantum yield of PSII photochemistry (Fv/Fm), ribulose-1,5-bisphosphate carboxylase (RuBisCO) activity, and starch content. In addition, PBZ treatment activated the antioxidant enzyme activities, antioxidants contents, ascorbate acid-glutathione (AsA-GSH) cycle, and related gene expression, lessening the ROS burst (H2O2 and O2â-). However, the gibberellic acid (GA) anabolism was remarkably decreased by PBZ treatment under LL stress, downregulating the transcript levels of kaurene oxidase (KO), kaurenoic acid oxidase (KAO), and GA 20-oxidases (GA20ox). At the same time, PBZ treatment up-regulated 9-cis-epoxycarotenoid dioxygenase (NCED) gene expression, significantly increasing the endogenous abscisic acid (ABA) concentration under LL stress. Thus, our study revealed that PBZ improves the antioxidation and photosynthetic capacity, meanwhile increasing the ABA concentration and decreasing GA concentration, which ultimately enhances the LL stress tolerance in tall fescue.
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Festuca , Lolium , Antioxidantes/farmacología , Clorofila/metabolismo , Festuca/metabolismo , Hormonas/metabolismo , Peróxido de Hidrógeno/metabolismo , Lolium/metabolismo , Fotosíntesis , Complejo de Proteína del Fotosistema II/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Triazoles/metabolismo , Triazoles/farmacologíaRESUMEN
The vital signaling molecule 5-Aminolevulinic acid (ALA) plays critical roles in signal transduction and biological modulation under abiotic stresses. In this study, we explored the effects of exogenous ALA on low-light (LL) stress-induced photosynthesis and antioxidant system damage in tall fescue (Festuca arundinacea Schreb.) seedlings. LL stress decreased morphological index values and chlorophyll contents, while also reduced net photosynthetic rate (Pn) and the maximum quantum yield of photosystem II photochemistry (Fv/Fm). Notably, these restrictions were substantially alleviated by exogenous ALA. Moreover, the contents of chlorophyll and its synthetic precursors were significantly increased after ALA treatment. Meanwhile, ALA observably enhanced expression level of FaCHLG, FaHEMA, FaPOR, and FaCAO, which encode the chlorophyll precursors biosynthesis enzymes. Exogenous ALA repaired the damage to the chloroplast ultrastructure caused by LL stress and promoted the formation of ordered thylakoids and grana lamella. ALA also improved Rubisco activity and expression level of the photosynthetic enzyme genes FaRuBP, FaPRK, and FaGADPH. Additionally, application of exogenous ALA decreased relative electrolytic leakage and the accumulation of malondialdehyde (MDA), hydrogen peroxide (H2O2), and superoxide radicals (O2â-), and increased the gene expression levels and activity of antioxidant enzymes. The ratios of ascorbic acid (AsA) to dehydroascorbic acid (DHA) and reduced glutathione (GSH) to oxidized glutathione (GSSG) were also increased significantly by application of ALA. Furthermore, all responses could be reversed by treatment with levulinic acid (LA). Thus, these results indicated that ALA protects tall fescue from LL stress through scavenging ROS, improving photosynthetic enzyme activity levels, increasing photosynthetic pigments contents, repairing chloroplast damage, and enhancing the photosynthesis rate.
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Ácido Aminolevulínico , Antioxidantes , Festuca , Ácido Aminolevulínico/metabolismo , Ácido Aminolevulínico/farmacología , Antioxidantes/metabolismo , Ácido Ascórbico/metabolismo , Clorofila/metabolismo , Cloroplastos/metabolismo , Ácido Deshidroascórbico/metabolismo , Festuca/metabolismo , Glutatión/metabolismo , Disulfuro de Glutatión/metabolismo , Disulfuro de Glutatión/farmacología , Peróxido de Hidrógeno/metabolismo , Malondialdehído/metabolismo , Fotosíntesis , Complejo de Proteína del Fotosistema II/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Ribulosa-Bifosfato Carboxilasa/metabolismo , Plantones/metabolismo , Superóxidos/metabolismoRESUMEN
Introduction: O-linked ß-D-N-acetylglucosamine (O-GlcNAc) modification is a post-translational modification in which a single O-GlcNAc is added to serine or threonine residues in nuclear, cytoplasmic, and mitochondrial proteins, and is involved in a variety of physiological processes. Objectives: In the present study, the role of O-GlcNAcylation in embryo implantation was evaluated. Furthermore, whether O-GlcNAcylation is involved in orchestrating glucose metabolism to influence endometrial cell physiological functions was investigated. Methods: Different endometrial tissues were detected using immunohistochemistry. Pregnant mouse models were established to verify molecular expression. O-GlcNAc transferase and aquaporin 3 (AQP3) knockdown were used to detect embryo implantation efficiency in vitro and in vivo. Western blotting and immunofluorescence were used to detect protein expression and stability. Dual luciferase reporter assay and chromatin immunoprecipitation (ChIP) were used to verify the binding transcription factor. Glycolysis was detected using bioenergy analyzer, and metabolites were analyzed using isotope 13C-labeled LC-MS. Metabolic-related genes were determined using RNA sequencing. Results: Activation of endometrial hexosamine biosynthetic pathway (HBP) caused elevated O-GlcNAcylation during the window of implantation, affecting endometrial cell function and embryo implantation. Specifically, elevated O-GlcNAcylation increased glucose uptake via glucose transporter 1 (GLUT1) leading to glucose metabolic flow into the pentose phosphate pathways and HBP, which regulate the metabolic reprogramming of endometrial cells. Furthermore, O-GlcNAcylation mediated the intracellular transport of glycerol to support and compensate for glycolysis through regulation of AQP3. Unexpectedly, elevated AQP3 also increased glucose uptake via GLUT1. These processes maintained higher metabolic requirements for endometrial physiology. Furthermore, the transcription factor SP1 specifically bound to the AQP3 promoter region, and O-GlcNAcylation of SP1 increased its stability and transcriptional regulation of AQP3 which is associated with O-GlcNAcylation of SP1. Conclusion: Overall, O-GlcNAcylation regulated glucose metabolism in endometrial cells, and AQP3-mediated compensation provides new insights into the communication between glycolysis and O-GlcNAcylation.
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Acuaporina 3 , Glucólisis , Animales , Acuaporina 3/metabolismo , Implantación del Embrión , Glucosa/metabolismo , Transportador de Glucosa de Tipo 1/metabolismo , Hexosaminas , RatonesRESUMEN
BACKGROUND: A hydatidiform mole is a condition caused by abnormal proliferation of trophoblastic cells. MicroRNA miR-30a acts as a tumor suppressor gene in most tumors and participates in the development of various cancers. However, its role in hydatidiform moles is not clear. METHODS: Quantitative real-time reverse transcription PCR was used to verify the expression level of miR-30a and STOX2 (encoding storkhead box 2). Flow cytometry assays were performed to detect the cell cycle in cell with different expression levels of miR-30a and STOX2. Cell Cycle Kit-8, 5-ethynyl-2'-deoxyuridine, and colony formation assays were used to detect cell proliferation and viability. Transwell assays was used to test cell invasion and migration. Dual-luciferase reporter assays and western blotting were used to investigate the potential mechanisms involved. RESULT: Low miR-30a expression promoted the proliferation, migration, and invasion of trophoblastic cells (JAR and HTR-8). Dual luciferase assays confirmed that STOX2 is a target of miR-30a and resisted the effect of upregulated miR-30a in trophoblastic cells. In addition, downregulation of STOX2 by miR-30a could activate ERK, AKT, and P38 signaling pathways. These results revealed a new mechanism by which ERK, AKT, and P38 activation by miR-30a/STOX2 results in excessive proliferation of trophoblast cells in the hydatidiform mole. CONCLUSIONS: In this study, we found that miR-30a plays an important role in the development of the hydatidiform mole. Our findings indicate that miR-30a might promote the malignant transformation of human trophoblastic cells by regulating STOX2, which strengthens our understanding of the role of miR-30a in regulating trophoblastic cell transformation.
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Hydrogen sulfide (H2S), as a gaseous messenger molecule, plays critical roles in signal transduction and biological modulation. In the present study, the roles of H2S in regulating chlorophyll (Chl) and carotenoid (Car) contents to improve photosynthesis in tall fescue were investigated under low-light (LL) stress. Compared to control conditions, LL stress significantly reduced total biomass, net photosynthetic rate (Pn), maximal quantum yield of photosystem II (PSII) photochemistry (Fv/Fm), and the contents of Chl and Car. Under exogenous sodium hydrosulfide (NaHS, H2S donor) application, these parameters were enhanced, ultimately increasing photosynthesis. Moreover, exogenous H2S up-regulated the expression of chlorophyll biosynthesis genes while down-regulated chlorophyll degradation genes, resulting in increases in chlorophyll precursors. Components of carotenoids and expression of genes encoding biosynthesis and degradation enzymes varied similarly. Additionally, application exogenous H2S up-regulated expression of FaDES1 and FaDCD. Thus, it enhanced L-cysteine desulfhydrase 1 (DES1, EC 4.4.1.1) and D-cysteine desulfhydrase (DCD, EC 4.4.1.15) activities leading to elevated endogenous H2S. However, these responses were reversed by treatment with hypotaurine (HT, H2S scavenger). These results suggested that H2S is involved in regulating photosynthesis to improve LL tolerance via modulating Chl and Car metabolisms in tall fescue.
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Festuca , Sulfuro de Hidrógeno , Carotenoides , Clorofila , FotosíntesisRESUMEN
The successful implantation of embryos is crucial for pregnancy in mammals. This complex process is inevitably dependent on the development of the endometrium. The paired-like homeodomain transcription factor 2 (PITX2) is involved in a variety of biological processes, but whether it is involved in embryo implantation has not been reported. In this study, we aimed to investigate uterine expression and regulation of PITX2 during implantation. We found that PITX2 was elevated in the human endometrium in the secretory phase. The results of the pregnant mouse models showed that PITX2 expression was spatiotemporal in mouse endometrial tissue throughout peri-implantation period, and it was significantly upregulated at the time of implantation. Interestingly, PITX2 was mainly localized to the glandular epithelium cells on D2.5-3.5 of pregnancy, while D5.5-6.5 was largely expressed in stromal cells. In vitro, PITX2 regulated endometrial cells proliferation, migration, invasion, and other functions through the Wnt/ß-catenin signaling pathway. In addition, a significant decrease in the rate of embryo implantation was observed after injecting PITX2 small interfering RNA into the uterine horn. These results demonstrate the effects of PITX2 on the physiological function of endometrial cells and embryo implantation, suggesting a role in the endometrial regulatory mechanism during implantation.
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Implantación del Embrión , Endometrio/metabolismo , Proteínas de Homeodominio/fisiología , Factores de Transcripción/fisiología , Vía de Señalización Wnt , Adulto , Animales , Línea Celular , Femenino , Humanos , Ratones , Ratones Endogámicos C57BL , Persona de Mediana Edad , Embarazo , Proteína del Homeodomínio PITX2RESUMEN
Acute kidney injury (AKI) is characterized by abrupt kidney dysfunction. It results in remote organ dysfunction, including the brain. The underlying mechanism of the kidneybrain axis in AKI and effective protective approaches remain unknown. The present study aimed to investigate the potential protective effect of ginsenoside (GS) on AKI induced by glycerol in rats. Kidney function was initially assessed by blood urea nitrogen (BUN) and creatinine (Cre) tests, and was identified to be severely impaired following glycerol treatment, based on significant increases in BUN and Cre levels observed. Severe extensive necrosis of the majority of the renal tubules was observed by hematoxylin and eosin staining, additionally confirming that glycerol induced AKI. GS was identified to ameliorate the impairment of kidney function in the context of AKI. Further investigation of the mechanism revealed that GS may induce protection against oxidative stress via a kidneybrain axis. Furthermore, GS improved the activation of hypoxiainducible factor 1α (HIF1α) and vascular endothelial growth factor A (VEGFA) in the hypothalamus response to AKI, and in the kidney tissues. The protective effect of GS in AKI may be associated with the interaction between the kidney and the brain. Taken together, these results suggested that GS was involved in the protective effects against AKI by decreasing oxidative damage to the kidney and brain, and by upregulating HIF1α and VEGFA levels in the kidneybrain axis.
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Lesión Renal Aguda/tratamiento farmacológico , Lesión Renal Aguda/metabolismo , Ginsenósidos/uso terapéutico , Factor A de Crecimiento Endotelial Vascular/metabolismo , Animales , Nitrógeno de la Urea Sanguínea , Western Blotting , Creatinina/metabolismo , Inmunohistoquímica , Riñón/efectos de los fármacos , Riñón/metabolismo , Masculino , Ratas , Ratas Sprague-DawleyRESUMEN
Olinked ßNacetylglucosamine (OGlcNAc) modification is a dynamic posttranslational modification process that is involved in many crucial biological processes, including cell cycle regulation, nutrient metabolism and extracellular signaling. This dynamic modification is dependent on the ambient glucose concentration and is catalyzed and removed by OGlcNAc transferase (OGT) and OGlcNAcase (OGA), respectively. The present study aimed to determine the role of OGlcNAcylation during embryo implantation by inhibiting or enhancing its function and expression. The results revealed that the expression of OGlcNAcmodified proteins in the human secretory endometrium was higher than that of the endometrium during the proliferative phase, as determined via western blotting and immunohistochemistry. Additionally, the level of endometrial OGlcNAc modification increased gradually from the prereceptive to the receptive phase, which was then decreased during the nonreceptive phase. In endometrial cells, RNA interference was utilized to reduce the expression of two key OGlcNAc synthesis and decomposition enzymes, OGT and OGA, to indirectly increase or decrease levels of OGlcNAc modification. The results revealed that increasing the level of OGlcNAc modification enhanced cellular proliferation, migration, invasion and adhesion, thereby promoting embryo implantation. It is hypothesized that OGlcNAc modification serves an important role in the regulation of endometrial receptivity and embryo implantation. The results of the present study may have important implications for the understanding of female fertility and may help improve infertility treatments.
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Acetilglucosamina/metabolismo , Antígenos de Neoplasias/metabolismo , Endometrio/metabolismo , Histona Acetiltransferasas/metabolismo , Hialuronoglucosaminidasa/metabolismo , N-Acetilglucosaminiltransferasas/metabolismo , Adulto , Antígenos de Neoplasias/genética , Línea Celular , Movimiento Celular , Proliferación Celular , Implantación del Embrión , Femenino , Fase Folicular/metabolismo , Glicosilación , Histona Acetiltransferasas/genética , Humanos , Hialuronoglucosaminidasa/genética , Persona de Mediana Edad , N-Acetilglucosaminiltransferasas/genéticaRESUMEN
Mycobacterium tuberculosis, a causative pathogen of tuberculosis (TB), still threatens human health worldwide. To find a novel drug to eradicate this pathogen, we tested taurine-5- bromosalicylaldehyde Schiff base (TBSSB) as an innovative anti-mycobacterial drug using Mycobacterium smegmatis as a surrogate model for M. tuberculosis. We investigated the antimicrobial activity of TBSSB against M. smegmatis by plotting growth curves, examined the effect of TBSSB on biofilm formation, observed morphological changes by scanning electron microscopy and transmission electron microscopy, and detected differentially expressed proteins using two-dimensional gel electrophoresis coupled with mass spectrometry. TBSSB inhibited mycobacterial growth and biofilm formation, altered cell ultrastructure and intracellular content, and inhibited cell division. Furthermore, M. smegmatis adapted itself to TBSSB inhibition by regulating the metabolic pathways and enzymatic activities of the identified proteins. NDMA-dependent methanol dehydrogenase, NAD(P)H nitroreductase, and amidohydrolase AmiB1 appear to be pivotal factors to regulate the M. smegmatis survival under TBSSB. Our dataset reinforced the idea that Schiff base-taurine compounds have the potential to be developed as novel anti-mycobacterial drugs.
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Aldehídos/farmacología , Antibacterianos/farmacología , Mycobacterium smegmatis/efectos de los fármacos , Proteómica , Bases de Schiff/farmacología , Taurina/análogos & derivados , Proteínas Bacterianas/efectos de los fármacos , Biopelículas/efectos de los fármacos , Biopelículas/crecimiento & desarrollo , Pared Celular/efectos de los fármacos , Redes y Vías Metabólicas/efectos de los fármacos , Mycobacterium smegmatis/crecimiento & desarrollo , Mycobacterium smegmatis/ultraestructura , Mycobacterium tuberculosis , Taurina/farmacologíaRESUMEN
Aberrant glycosylation of proteins has major implications for human diseases. To determine whether protein glycosylation contributes to the pathogenesis of Parkinson's disease (PD), a mouse model of PD was established by injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Induction of PD-like features was verified by assessing motor impairment and confirming reductions in biological markers, including dopamine, 5-hydroxytryptamine and tyrosine hydroxylase, as well as the aggregation of α-synuclein. Altered glycosylation was detected using biotinylated agaracus bisporus lectin, which specifically binds exposed Gal-(ß-1,3)-GalNAc linked to glycoproteins. Subsequent lectin affinity chromatography coupled with mass spectrometry revealed enhanced glycosylation of microtubule-associated protein 6 (MAP6) in PD mice as compared to healthy controls. In situ dual co-immunofluorescence analysis and immunoblotting confirmed that MAP6 is glycosylated with Gal-(ß-1,3)-GalNAc oligosaccharides, which in turn alters the distribution and structure of MAP6 complexes within neurons. This is the first study to described MAP6 as a glycoprotein containing Gal-(ß-1,3)-GalNAc oligosaccharides and to show that hyperglycosylation of MAP6 is strongly associated with the pathogenesis of PD. These findings provide potentially valuable information for developing new therapeutic targets for the treatment of PD as well as reliably prognostic biomarkers.
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Galectinas/metabolismo , Proteínas Asociadas a Microtúbulos/metabolismo , Oligosacáridos/metabolismo , Enfermedad de Parkinson/metabolismo , 1-Metil-4-fenil-1,2,3,6-Tetrahidropiridina , Animales , Cromatografía de Afinidad , Modelos Animales de Enfermedad , Técnica del Anticuerpo Fluorescente , Glicoproteínas/metabolismo , Glicosilación , Immunoblotting , Masculino , Ratones , Ratones Endogámicos C57BL , Enfermedad de Parkinson/patología , Espectrometría de Masa por Láser de Matriz Asistida de Ionización DesorciónRESUMEN
Functionally uncharacterized gene Rv3627c is predicted to encode a carboxypeptidase in the pathogen of Mycobacterium tuberculosis (M. tuberculosis), which remains a major threat to human health. Here, we sought to reveal the function of Rv3627c and to elucidate its effects on mycobacterial growth. Rv3627c was purified from E. coli using Ni2+-NTA affinity chromatography, and its identity was confirmed with a monoclonal anti-polyhistidine antibody. An enzyme activity assay involving a d-amino acid oxidase-peroxidase coupled colorimetric reaction and high-performance thin layer chromatography was performed. A pull-down assay and MS-MS were also employed to identify putative interaction partners of Rv3627c. Scanning electron microscopy and transmission electron microscopy were performed to observe any morphological alterations to Mycobacterium smegmatis (M. smegmatis). We successfully obtained soluble expressed Rv3627c and identified it as carboxypeptidase using prepared peptidoglycan. Four proteins were identified as potential interaction partners with Rv3627c based on results obtained from both a pull-down assay and MS/MS analysis. Rv3627c over-expression induced M. smegmatis cells to become elongated, and promoted the formation of increased numbers of Z-rings. Rv3627c, a novel carboxypeptidase in M. tuberculosis identified in this study, exerts important effects on mycobacterial cell morphology and cell division. This functional information provides a promising insight into anti-mycobacterial target designs.
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Proteínas Bacterianas/metabolismo , Carboxipeptidasas/metabolismo , División Celular , Mycobacterium smegmatis/citología , Mycobacterium smegmatis/enzimología , Secuencia de Aminoácidos , Proteínas Bacterianas/genética , Carboxipeptidasas/genética , Clonación Molecular , Dipéptidos/metabolismo , Humanos , Mapas de Interacción de Proteínas , Homología de SecuenciaRESUMEN
Ribonuclease inhibitor (RI), also termed angiogenin inhibitor, acts as the inhibitor of ribonucleolytic activity of RNase A and angiogenin. The expression of RI has been investigated in melanoma and bladder cancer cells. However, the precise role of RI in tumorigenesis, in addition to RIinduced autophagy, remains poorly understood. In the present study, it was demonstrated that RI positively regulated autophagy in human colorectal cancer (CRC) cells as indicated by an increase in light chain 3 (LC3)II levels. Furthermore, RI regulated cell survival in HT29 cells. In addition, autophagyassociated proteins, including beclin1 and autophagyrelated protein 13, were increased in response to RIinduced autophagy, and the protein kinase B (Akt)/mechanistic target of rapamycin (mTOR)/Unc51 like autophagy activating kinase (ULK1) pathway may be involved in the activation of autophagy induced by RI overexpression. Taken together, the evidence of the present study indicated that the overexpression of RI induced ATGdependent autophagy in CRC cells via the Akt/mTOR/ULK1 pathway, suggesting that the upregulation of RI activity may constitute a novel approach for the treatment of human colorectal carcinoma.