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Bronchopulmonary dysplasia (BPD) is a chronic lung disease of prematurity. Exposure to noxious stimuli such as hyperoxia, volutrauma, and infection in infancy can have long-reaching impacts on lung health and predispose towards the development of conditions such as chronic obstructive pulmonary disease (COPD) in adulthood. BPD and COPD are both marked by lung tissue degradation, neutrophil influx, and decreased lung function. Both diseases also express a change in microbial signature characterized by firmicute depletion. However, the relationship between pulmonary bacteria and the mechanisms of downstream disease development has yet to be elucidated. We hypothesized that murine models of BPD would show heightened acetylated proline-glycine-proline (Ac-PGP) pathway and neutrophil activity, and through gain- and loss-of-function studies we show that Ac-PGP plays a critical role in driving BPD development. We further test a inhaled live biotherapeutic (LBP) using active Lactobacillus strains in in vitro and in vivo models of BPD and COPD. The Lactobacillus-based LBP is effective in improving lung structure and function, mitigating neutrophil influx, and reducing a broad swath of pro-inflammatory markers in these models of chronic pulmonary disease via the MMP-9/PGP (matrix metalloproteinase/proline-glycine-proline) pathway. Inhaled LBPs show promise in addressing common pathways of disease progression that in the future can be targeted in a variety of chronic lung diseases.
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Displasia Broncopulmonar , Modelos Animais de Doenças , Lactobacillus , Pulmão , Neutrófilos , Doença Pulmonar Obstrutiva Crônica , Animais , Neutrófilos/imunologia , Camundongos , Administração por Inalação , Humanos , Doença Pulmonar Obstrutiva Crônica/imunologia , Doença Pulmonar Obstrutiva Crônica/microbiologia , Doença Pulmonar Obstrutiva Crônica/terapia , Pulmão/imunologia , Pulmão/patologia , Pulmão/microbiologia , Displasia Broncopulmonar/imunologia , Displasia Broncopulmonar/microbiologia , Camundongos Endogâmicos C57BL , Feminino , Probióticos/administração & dosagem , Probióticos/uso terapêutico , Pneumonia/microbiologia , Pneumonia/imunologia , Masculino , Prolina/metabolismoRESUMO
Surface plasmon polaritons (SPPs) have become a research hotspot due to their high intensity and subwavelength localization. Through free-electron excitation, a portion of the momentum of moving electrons can be converted into SPPs. Converting highly localized SPPs into a radiated field is an approach with the potential to aid in the development of a light radiation source. Reducing losses of SPPs is currently a critical challenge that needs to be addressed. The lifetime of SPPs in metal films is longer than that in metal blocks. Traditional optical gratings can transform SPPs into radiation to avoid the decay of SPPs in metal; however, they are created by etching metal films, so they tend to alter the dispersion characteristics of these films and will emit radiation in the direction perpendicular to the metal surface. This paper proposes an approach to converting the SPPs of a metal film excited by free electrons into a radiation field via lateral grating and obtaining in-plane radiation. We investigate the properties of SPP lateral radiation. The study of lateral radiation from metal films holds significant importance for SPP radiation sources and SPP on-chip circuit development.
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In this paper, we use the method of high order TMn1 mode selection from the concept of narrow-band Smith-Purcell radiation (SPR) for powerful, over-mode, multi-gap extended interaction circuit designs toward millimeter wave and Terahertz (THz) region. As a core part, the multiple gaps interaction structure, equivalent to a subwavelength hole array (SHA), excites the narrow band SPR when an electron beam is injected. The SPR energy is collected by a pair of closed cavities, which satisfies (n-1) standing wave units. The SPR energy in the optimized cavity allows a high index n TMn1 mode operation to achieve the strongest Ez field and high characteristic impedance in a closed multi-gap resonant circuit. This provides an effective design to establish a stable high-order TMn1 mode that supports extended interaction circuits with large cross sections. A 0.46 THz extended interaction circuit, employing the novel high order TM51-2π mode operation output structure, has been designed to demonstrate the efficient beam-wave interaction in the proposed system. The method of TMn1 mode selection provides new insight into the understanding of the high-frequency extended interaction circuits by introducing the SPR concept, benefiting the development of millimeter wave and THz vacuum electron devices (VEDs).
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Vascular calcification is accelerated in patients with diabetes mellitus and increases risk of cardiovascular events and mortality. Vascular smooth muscle cells (VSMC) play a key role in regulating vascular tone and contribute significantly to the development of diabetic vasculopathy. In this study, the function of stromal interaction molecule 1 (STIM1), an important regulator for intracellular calcium homeostasis, in diabetic vascular calcification was investigated, and the underlying molecular mechanisms were uncovered. A SMC-specific STIM1 deletion mouse model (STIM1Δ/Δ) was generated by breeding the STIM1 floxed mice (STIM1f/f) with SM22α-Cre transgenic mice. Using aortic arteries from the STIM1Δ/Δ mice and their STIM1f/f littermates, we found that SMC-specific STIM1 deletion induced calcification of aortic arteries cultured in osteogenic media ex vivo. Furthermore, STIM1 deficiency promoted osteogenic differentiation and calcification of VSMC from the STIM1Δ/Δ mice. In the low-dose streptozotocin (STZ)-induced mouse model of diabetes, SMC-specific STIM1 deletion markedly enhanced STZ-induced vascular calcification and stiffness in the STIM1Δ/Δ mice. The diabetic mice with SMC-specific STIM1 ablation also exhibited increased aortic expression of the key osteogenic transcription factor, Runx2, and protein O-GlcNAcylation, an important post-translational modulation that we have reported to promote vascular calcification and stiffness in diabetes. Consistently, elevation of O-GlcNAcylation was demonstrated in aortic arteries and VSMC from the STIM1Δ/Δ mice. Inhibition of O-GlcNAcylation with a pharmacological inhibitor abolished STIM1 deficiency-induced VSMC calcification, supporting a critical role of O-GlcNAcylation in mediating STIM1 deficiency-induced VSMC calcification. Mechanistically, we identified that STIM1 deficiency resulted in impaired calcium homeostasis, which activated calcium signaling and increased endoplasmic reticulum (ER) stress in VSMC, while inhibition of ER stress attenuated STIM1-induced elevation of protein O-GlcNAcylation. In conclusion, the study has demonstrated a causative role of SMC-expressed STIM1 in regulating vascular calcification and stiffness in diabetes. We have further identified a novel mechanisms underlying STIM1 deficiency-induced impairment of calcium homeostasis and ER stress in upregulation of protein O-GlcNAcylation in VSMC, which promotes VSMC osteogenic differentiation and calcification in diabetes.
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Diabetes Mellitus Experimental , Calcificação Vascular , Camundongos , Animais , Cálcio/metabolismo , Diabetes Mellitus Experimental/metabolismo , Osteogênese , Células Cultivadas , Calcificação Vascular/etiologia , Camundongos Transgênicos , Modelos Animais de Doenças , Homeostase , Miócitos de Músculo Liso/metabolismoRESUMO
BACKGROUND: MiT-Renal Cell Carcinoma (RCC) is characterized by genomic translocations involving microphthalmia-associated transcription factor (MiT) family members TFE3, TFEB, or MITF. MiT-RCC represents a specific subtype of sporadic RCC that is predominantly seen in young patients and can present with heterogeneous histological features making diagnosis challenging. Moreover, the disease biology of this aggressive cancer is poorly understood and there is no accepted standard of care therapy for patients with advanced disease. Tumor-derived cell lines have been established from human TFE3-RCC providing useful models for preclinical studies. METHODS: TFE3-RCC tumor derived cell lines and their tissues of origin were characterized by IHC and gene expression analyses. An unbiased high-throughput drug screen was performed to identify novel therapeutic agents for treatment of MiT-RCC. Potential therapeutic candidates were validated in in vitro and in vivo preclinical studies. Mechanistic assays were conducted to confirm the on-target effects of drugs. RESULTS: The results of a high-throughput small molecule drug screen utilizing three TFE3-RCC tumor-derived cell lines identified five classes of agents with potential pharmacological efficacy, including inhibitors of phosphoinositide-3-kinase (PI3K) and mechanistic target of rapamycin (mTOR), and several additional agents, including the transcription inhibitor Mithramycin A. Upregulation of the cell surface marker GPNMB, a specific MiT transcriptional target, was confirmed in TFE3-RCC and evaluated as a therapeutic target using the GPNMB-targeted antibody-drug conjugate CDX-011. In vitro and in vivo preclinical studies demonstrated efficacy of the PI3K/mTOR inhibitor NVP-BGT226, Mithramycin A, and CDX-011 as potential therapeutic options for treating advanced MiT-RCC as single agents or in combination. CONCLUSIONS: The results of the high-throughput drug screen and validation studies in TFE3-RCC tumor-derived cell lines have provided in vitro and in vivo preclinical data supporting the efficacy of the PI3K/mTOR inhibitor NVP-BGT226, the transcription inhibitor Mithramycin A, and GPNMB-targeted antibody-drug conjugate CDX-011 as potential therapeutic options for treating advanced MiT-RCC. The findings presented here should provide the basis for designing future clinical trials for patients with MiT-driven RCC.
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Carcinoma de Células Renais , Neoplasias Renais , Humanos , Carcinoma de Células Renais/patologia , Neoplasias Renais/patologia , Inibidores de MTOR , Fatores de Transcrição de Zíper de Leucina e Hélice-Alça-Hélix Básicos/genética , Translocação Genética , Fosfatidilinositol 3-Quinase , Glicoproteínas de Membrana/genéticaRESUMO
Angiotensin (1-7) [Ang (1-7)] is an active heptapeptide of the noncanonical arm of the renin-angiotensin system that modulates molecular signaling pathways associated with vascular and cellular inflammation, vasoconstriction, and fibrosis. Preclinical evidence suggests that Ang (1-7) is a promising therapeutic target that may ameliorate physical and cognitive function in late life. However, treatment pharmacodynamics limits its clinical applicability. Therefore, this study explored the underlying mechanisms altered by a genetically modified probiotic (GMP) that expresses Ang (1-7) combined with and without exercise training in an aging male rat model as a potential adjunct strategy to exercise training to counteract the decline of physical and cognitive function. We evaluated cross-tissue (prefrontal cortex, hippocampus, colon, liver, and skeletal muscle) multi-omics responses. After 12 wk of intervention, the 16S mRNA microbiome analysis revealed a main effect of probiotic treatment within- and between groups. The probiotic treatment enhanced α diversity (Inverse Simpson (F[2,56] = 4.44; P = 0.02); Shannon-Wiener (F[2,56] = 4.27; P = 0.02)) and ß-diversity (F[2,56] = 2.66; P = 0.01) among rats receiving our GMP. The analysis of microbes' composition revealed three genera altered by our GMP (Enterorhabdus, Muribaculaceae unclassified, and Faecalitalea). The mRNA multi-tissue data analysis showed that our combined intervention upregulated neuroremodeling pathways on prefrontal cortex (i.e., 140 genes), inflammation gene expression in the liver (i.e., 63 genes), and circadian rhythm signaling on skeletal muscle. Finally, the integrative network analysis detected different communities of tightly (|r| > 0.8 and P < 0.05) correlated metabolites, genera, and genes in these tissues.NEW & NOTEWORTHY This manuscript uses a multiomics approach (i.e., microbiome, metabolomics, and transcriptomics) to explore the underlying mechanisms driven by a genetically modified probiotic (GMP) designed to express angiotensin (1-7) combined with moderate exercise training in an aged male rat model. After 12 wk of intervention, our findings suggest that our GMP enhanced gut microbial diversity while exercise training altered the transcriptional response in relevant neuroremodeling genes, inflammation, and circadian rhythm signaling pathways in an aging animal model.
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Multiômica , Condicionamento Físico Animal , Ratos , Animais , Masculino , Condicionamento Físico Animal/fisiologia , Sistema Renina-Angiotensina/fisiologia , InflamaçãoRESUMO
Age-related declines in physical and cognitive function can have tremendous, negative impacts on health span and quality of life. Therefore, we investigated the potential of utilizing a probiotic treatment to target the renin-angiotensin system (RAS) in conjunction with moderate exercise to ameliorate age-related declines in cognitive and physical function in aged rats. Herein we utilized a genetically modified angiotensin (1-7), which activates a "complementary" arm of the RAS through binding Mas (AT7) receptors. This process induces several beneficial physiologic effects, including decreased inflammation and enhanced physical/cognitive function. Thus, in this short research report, we suggest the efficacy of this Ang(1-7) releasing Lactobacillus paracasei (LPA) as either an alternative strategy to exercise, or more likely as an adjuvant to moderate exercise, for the prevention of both physical and cognitive decline especially in female rats.
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Angiotensina II , Qualidade de Vida , Feminino , Ratos , Animais , Sistema Renina-Angiotensina/fisiologia , Angiotensina I , Fragmentos de PeptídeosRESUMO
INTRODUCTION: Growing research suggests that aerobic high-intensity interval training (HIIT) improves cardiovascular function and physical performance compared with moderate intensity continuous training (MICT). However relatively few animal models of HIIT are available to inform about the benefits of this exercise-particularly among older animals. In addition, there is little evidence for how HIIT training interacts with adjuvant pharmacological therapies known to enhance the impact of MCIT in older individuals such as Angiotensin Converting Enzyme (ACE) Inhibitors. PURPOSE: The aim of the present study was to establish a HIIT protocol in aged rats based on forced running wheel-bed, and to subsequently (1) establish the feasibility of the HIIT protocol in a proof-of-concept study evaluating interactions between HIIT and (2) the result of combining HIIT + ACE inhibitor treatment using the ACE inhibitor enalapril. METHODS: Two groups of rats were used in this study. The feasibility of using wheel-bed for HIIT training was tested in group one (15- and 30-month-old male rats). In the second group, 37 24-month-old Fisher 344 × Brown Norway male rats were randomly divided into four subgroups: control, enalapril, HIIT training group, and HIIT training combined with enalapril administration. The training and administration lasted for 4 weeks. After the intervention, locomotor activity, exercise tolerance, and grip strength were tested. RESULTS: Our feasibility study suggested that middle-aged and aged rats were able to successfully complete the HIIT training. In our intervention study, HIIT training alone, regardless of adjuvant enalapril intervention, did raise treadmill exercise tolerance vs. the sedentary condition. Measures of healthspan were not negatively impacted by HIIT training. CONCLUSION: The novel HIIT protocol based on forced running wheel-bed was successfully employed in aged rats. We conclude that future studies should compare the results and of multi-modal intervention strategies which include both HIIT and MICT in combination with adjuvant therapies such as enalapril to improve exercise tolerance and other global indices of healthspan.
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ABSTRACT It is particularly important to improve the quality of physical education curriculum and the overall health level of college students. It is difficult to achieve a comprehensive evaluation of the original evaluation method of students' physical health level. Therefore, this study combined the analytic hierarchy process (AHP) and physical fitness training mode to build a university physical health evaluation model. Taking X University as the experimental background, 20 students of different majors in grade 2019 were selected as experimental objects. One group (10 students) adopted the original physical education curriculum mode, and the other group (10 students) integrated physical fitness training into the physical education curriculum. Using the established health evaluation model, the evaluation scores of physical health level of two groups of college students before and after the experiment were comprehensively analyzed. The results showed that the overall score of group A students' physical health level increased; the rising rate was 2.87%. However, there was no significant change in the scores of physical health assessment of group B students. The results show that physical fitness training into the public physical education curriculum can effectively help improve the overall health level of students, and also shows that the introduction of AHP into the evaluation of College Students' physical health level can improve the comprehensiveness of health level analysis. It is hoped that this study can provide some reference for physical education teaching in Colleges and universities, so as to improve the overall teaching level of colleges and universities.
RESUMO É particularmente importante melhorar a qualidade dos programas de educação física e o nível geral de saúde dos estudantes universitários. É difícil obter uma avaliação abrangente do método de avaliação inicial do nível de saúde física dos estudantes. Por conseguinte, este estudo combinou o processo hierárquico analítico (AHP) e o modo de condicionamento físico para construir um modelo universitário de avaliação da saúde física. Tendo a Universidade X como objeto experimental, vinte estudantes de diferentes especializações do período de 2019 foram selecionados como objetos experimentais. Um grupo ( de dez estudantes) adotou o modo de educação física original, e o outro grupo (de dez estudantes) integrou o condicionamento físico no currículo de educação física. Utilizando o modelo estabelecido de avaliação da saúde, as pontuações de avaliação do nível de saúde física de dois grupos de estudantes universitários antes e depois do experimento foram exaustivamente analisadas. Os resultados mostraram que a pontuação global do nível de saúde física de um grupo de estudantes aumentou. A taxa de aumento foi de 2.87%. No entanto, não houve alterações significativas nas pontuações da avaliação para a saúde física dos estudantes do grupo B. Os resultados mostram que o condicionamento físico no currículo público de educação física pode contribuir efetivamente para melhorar o nível geral de saúde dos estudantes, e também mostra que a introdução do processo hierárquico analítico (AHP) na avaliação do nível de saúde física dos estudantes universitários pode melhorar a abrangência da análise do nível de saúde. Espera-se que este estudo possa fornecer uma referência para o ensino de educação física nos colégios e universidades a fim de melhorar o nível de ensino geral das universidades.
RESUMEN Es particularmente importante mejorar la calidad del plan de estudios de educación física y el nivel de salud general de los estudiantes universitarios. Es difícil lograr un análisis integral del método de evaluación original del nivel de salud física de los estudiantes. Por lo tanto, este estudio combinó el proceso de jerarquía analítica (PJA) y el modo de entrenamiento de aptitud física para construir un modelo universitario de evaluación de la salud física. Tomando la Universidad X como base experimental, se seleccionaron como sujetos experimentales 20 estudiantes de diferentes carreras en los cursos de 2019. Un grupo (10 estudiantes) adoptó el modo de plan de estudios de educación física original y el otro grupo (10 estudiantes) integró el entrenamiento de aptitud física en el plan de estudios de educación física. Utilizando el modelo de evaluación de salud establecido, se analizaron exhaustivamente los puntajes de evaluación del nivel de salud física de los dos grupos de estudiantes universitarios antes y después del experimento. Los resultados mostraron que el puntaje general del nivel de salud física de los estudiantes del grupo A aumentó; la tasa de incremento fue del 2,87%. Sin embargo, no hubo cambios significativos en las puntuaciones de la evaluación de la salud física de los estudiantes del grupo B. Los resultados muestran que el entrenamiento de la aptitud física en el plan de estudios público de educación física puede ayudar de manera efectiva a mejorar el nivel de salud general de los estudiantes y también muestra que la introducción de PJA en la evaluación del nivel de salud física de los estudiantes universitarios puede mejorar la precisión del análisis del nivel de salud. Esperamos que este estudio pueda proporcionar una referencia para la enseñanza de la educación física en los colegios y universidades, a fin de mejorar su nivel general en estos establecimientos.
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Humanos , Educação Física e Treinamento , Esportes/educação , Universidades , Nível de Saúde , Currículo , Condicionamento Físico HumanoRESUMO
Renal cell carcinoma (RCC) is not a single disease but is made up of several different histologically defined subtypes that are associated with distinct genetic alterations which require subtype specific management and treatment. Papillary renal cell carcinoma (pRCC) is the second most common subtype after conventional/clear cell RCC (ccRCC), representing ~20% of cases, and is subcategorized into type 1 and type 2 pRCC. It is important for preclinical studies to have cell lines that accurately represent each specific RCC subtype. This study characterizes seven cell lines derived from both primary and metastatic sites of type 1 pRCC, including the first cell line derived from a hereditary papillary renal carcinoma (HPRC)-associated tumor. Complete or partial gain of chromosome 7 was observed in all cell lines and other common gains of chromosomes 16, 17, or 20 were seen in several cell lines. Activating mutations of MET were present in three cell lines that all demonstrated increased MET phosphorylation in response to HGF and abrogation of MET phosphorylation in response to MET inhibitors. CDKN2A loss due to mutation or gene deletion, associated with poor outcomes in type 1 pRCC patients, was observed in all cell line models. Six cell lines formed tumor xenografts in athymic nude mice and thus provide in vivo models of type 1 pRCC. These type 1 pRCC cell lines provide a comprehensive representation of the genetic alterations associated with pRCC that will give insight into the biology of this disease and be ideal preclinical models for therapeutic studies.
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Carcinoma de Células Renais/genética , Autenticação de Linhagem Celular/métodos , Neoplasias Renais/genética , Ensaios Antitumorais Modelo de Xenoenxerto/métodos , Animais , Carcinoma de Células Renais/patologia , Linhagem Celular Tumoral , Instabilidade Cromossômica , Inibidor p16 de Quinase Dependente de Ciclina/genética , Inibidor p16 de Quinase Dependente de Ciclina/metabolismo , Humanos , Neoplasias Renais/patologia , Camundongos , Mutação , Proteínas Proto-Oncogênicas c-met/genética , Proteínas Proto-Oncogênicas c-met/metabolismoRESUMO
Understanding the mechanisms of the Warburg shift to aerobic glycolysis is critical to defining the metabolic basis of cancer. Hereditary leiomyomatosis and renal cell carcinoma (HLRCC) is an aggressive cancer characterized by biallelic inactivation of the gene encoding the Krebs cycle enzyme fumarate hydratase, an early shift to aerobic glycolysis, and rapid metastasis. We observed impairment of the mitochondrial respiratory chain in tumors from patients with HLRCC. Biochemical and transcriptomic analyses revealed that respiratory chain dysfunction in the tumors was due to loss of expression of mitochondrial DNA (mtDNA)-encoded subunits of respiratory chain complexes, caused by a marked decrease in mtDNA content and increased mtDNA mutations. We demonstrated that accumulation of fumarate in HLRCC tumors inactivated the core factors responsible for replication and proofreading of mtDNA, leading to loss of respiratory chain components, thereby promoting the shift to aerobic glycolysis and disease progression in this prototypic model of glucose-dependent human cancer.
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Carcinoma de Células Renais/genética , Ciclo do Ácido Cítrico , Dano ao DNA , DNA Mitocondrial/metabolismo , Fumarato Hidratase/genética , Neoplasias Renais/genética , Leiomiomatose/enzimologia , Síndromes Neoplásicas Hereditárias/enzimologia , Neoplasias Cutâneas/enzimologia , Neoplasias Uterinas/enzimologia , Adulto , Idoso , Carcinoma de Células Renais/etiologia , Carcinoma de Células Renais/metabolismo , Reparo do DNA , Replicação do DNA , Feminino , Fumarato Hidratase/deficiência , Perfilação da Expressão Gênica , Humanos , Neoplasias Renais/etiologia , Neoplasias Renais/metabolismo , Leiomiomatose/complicações , Masculino , Pessoa de Meia-Idade , Mitocôndrias/genética , Mitocôndrias/metabolismo , Mutação , Síndromes Neoplásicas Hereditárias/complicações , Neoplasias Cutâneas/complicações , Neoplasias Uterinas/complicações , Adulto JovemRESUMO
Myeloid-derived suppressor cells (MDSC) and tumor-associated macrophages (TAM) contribute to cancer-related inflammation and tumor progression. While several myeloid molecules have been ascribed a regulatory function in these processes, the triggering receptors expressed on myeloid cells (TREMs) have emerged as potent modulators of the innate immune response. While various TREMs amplify inflammation, others dampen it and are emerging as important players in modulating tumor progression-for instance, soluble TREM-1 (sTREM-1), which is detected during inflammation, associates with disease progression, while TREM-2 expression is associated with tumor-promoting macrophages. We hypothesized that TREM-1 and TREM-2 might be co-expressed on tumor-infiltrating myeloid cells and that elevated sTREM-1 associates with disease outcomes, thus representing a possibility for mutual modulation in cancer. Using the 4T1 breast cancer model, we found TREM-1 and TREM-2 expression on MDSC and TAM and that sTREM-1 was elevated in tumor-bearing mice in multiple models and correlated with tumor volume. While TREM-1 engagement enhanced TNF, a TREM-2 ligand was detected on MDSC and TAM, suggesting that both TREM could be functional in the tumor setting. Similarly, we detected TREM-1 and Trem2 expression in myeloid cells in the RENCA model of renal cell carcinoma (RCC). We confirmed these findings in human disease by demonstrating the expression of TREM-1 on tumor-infiltrating myeloid cells from patients with RCC and finding that sTREM-1 was increased in patients with RCC. Finally, The Cancer Genome Atlas analysis shows that TREM1 expression in tumors correlates with poor outcomes in RCC. Taken together, our data suggest that manipulation of the TREM-1/TREM-2 balance in tumors may be a novel means to modulate tumor-infiltrating myeloid cell phenotype and function.
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PURPOSE: To evaluate the safety and feasibility of combining exercise (EX) and resveratrol to treat older adults with physical function limitations. METHODS: Three-arm, two-site pilot randomized, controlled trial (RCT) for community-dwelling adults (N = 60), 71.8 ± 6.3 years of age with functional limitations. Participants were randomized to receive either 12 weeks of (1) EX + placebo [EX0], (2) EX + 500 mg/day resveratrol [EX500], or (3) EX + 1000 mg/day resveratrol [EX1000]. EX consisted of two sessions a week for 12 weeks of center-based walking and whole-body resistance training. Safety was assessed through adverse events and feasibility through exercise session and supplement (placebo, or resveratrol) protocol adherence. Outcome measures included a battery of indices of physical function as well as skeletal muscle mitchondrial function. Data were adjusted for age and gender using the Intent-To-Treat approach. RESULTS: Adverse event frequency and type were similar between groups (n = 8 EX0, n = 12 EX500, and n = 7 EX1000). Overall, 85% of participants met the supplement adherence via pill counts while 82% met the exercise session adherence. Adjusted within group mean differences (95% confidence interval) from week 0 to 12 for gait speed ranged from -0.04 (EX0: -0.1, 0.03) m/s to 0.04 (EX1000: -0.02, 0.11) and the six-minute walk test mean differences were 9.45 (EX0: -9.02, 27.7), 22.9 (EX500: 4.18, 41.6), and 33.1 (EX1000: 13.8, 52.4) meters. Unadjusted mean differences for citrate synthase were -0.80 (EX0: -15.45, 13.84), -1.38 (EX500: -12.16, 9.39), and 7.75 (EX1000: -4.68, 20.18) mU/mg. COX activity mean within group changes ranged from -0.05 (EX0) to 0.06 (EX500) k/s/mg. Additional outcomes are detailed in the text. CONCLUSION: The pilot RCT indicated that combined EX + resveratrol was safe and feasible for older adults with functional limitations and may improve skeletal muscle mitochondrial function and mobility-related indices of physical function. A larger trial appears warranted and is needed to formally test these hypotheses.
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Exercício Físico , Caminhada , Idoso , Terapia por Exercício , Humanos , Projetos Piloto , ResveratrolRESUMO
An important context in which metabolism influences tumorigenesis is the genetic cancer syndrome hereditary leiomyomatosis and renal cell carcinoma (HLRCC), a disease in which mutation of the tricarboxylic acid cycle enzyme fumarate hydratase (FH) causes hyperaccumulation of fumarate. This electrophilic oncometabolite can alter gene activity at the level of transcription, via reversible inhibition of epigenetic dioxygenases, as well as posttranslationally, via covalent modification of cysteine residues. To better understand the potential for metabolites to influence posttranslational modifications important to tumorigenesis and cancer cell growth, here we report a chemoproteomic analysis of a kidney-derived HLRCC cell line. Using a general reactivity probe, we generated a data set of proteomic cysteine residues sensitive to the reduction in fumarate levels caused by genetic reintroduction of active FH into HLRCC cell lines. This revealed a broad up-regulation of cysteine reactivity upon FH rescue, which evidence suggests is caused by an approximately equal proportion of transcriptional and posttranslational modification-mediated regulation. Gene ontology analysis highlighted several new targets and pathways potentially modulated by FH mutation. Comparison of the new data set with prior studies highlights considerable heterogeneity in the adaptive response of cysteine-containing proteins in different models of HLRCC. This is consistent with emerging studies indicating the existence of cell- and tissue-specific cysteine-omes, further emphasizing the need for characterization of diverse models. Our analysis provides a resource for understanding the proteomic adaptation to fumarate accumulation and a foundation for future efforts to exploit this knowledge for cancer therapy.
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Cisteína/metabolismo , Fumarato Hidratase/metabolismo , Fumaratos/metabolismo , Neoplasias Renais/metabolismo , Leiomiomatose/metabolismo , Síndromes Neoplásicas Hereditárias/metabolismo , Neoplasias Cutâneas/metabolismo , Neoplasias Uterinas/metabolismo , Linhagem Celular Tumoral , Cisteína/genética , Fumarato Hidratase/genética , Humanos , Neoplasias Renais/genética , Neoplasias Renais/patologia , Leiomiomatose/genética , Leiomiomatose/patologia , Síndromes Neoplásicas Hereditárias/genética , Síndromes Neoplásicas Hereditárias/patologia , Neoplasias Cutâneas/genética , Neoplasias Cutâneas/patologia , Neoplasias Uterinas/genética , Neoplasias Uterinas/patologiaRESUMO
BACKGROUND: The loss-of-function mutation of fumarate hydratase (FH) is a driver of hereditary leiomyomatosis and renal cell carcinoma (HLRCC). Fumarate accumulation results in activation of stress-related mechanisms leading to upregulation of cell survival-related genes. To better understand how cells compensate for the loss of FH in HLRCC, we determined the amino acid nutrient requirements of the FH-deficient UOK262 cell line (UOK262) and its FH-repleted control (UOK262WT). METHODS: We determined growth rates and survival of cell lines in response to amino acid depletion and supplementation. RNAseq was used to determine the transcription changes contingent on Asn and Gln supplementation, which was further followed with stable isotope resolved metabolomics (SIRM) using both [U- 13C,15N] Gln and Asn. RESULTS: We found that Asn increased the growth rate of both cell lines in vitro. Gln, but not Asn, increased oxygen consumption rates and glycolytic reserve of both cell lines. Although Asn was taken up by the cells, there was little evidence of Asn-derived label in cellular metabolites, indicating that Asn was not catabolized. However, Asn strongly stimulated Gln labeling of uracil and precursors, uridine phosphates and hexosamine metabolites in the UOK262 cells and to a much lesser extent in the UOK262WT cells, indicating an activation of the hexosamine biosynthetic pathway (HBP) by Asn. Asn in combination with Gln, but not Asn or Gln alone, stimulated expression of genes associated with the endoplasmic reticulum (ER) stress and the unfolded protein response (UPR) in UOK262 to a greater extent than in FH-restored cells. The changes in expression of these genes were confirmed by RT-PCR, and the stimulation of the UPR was confirmed orthogonally by demonstration of an increase in spliced XBP1 (sXBP1) in UOK262 cells under these conditions. Asn exposure also increased both the RNA and protein expression of the HBP regulator GFPT2, which is a transcriptional target of sXBP1. CONCLUSIONS: Asn in the presence of Gln induces an ER stress response in FH-deficient UOK262 cells and stimulates increased synthesis of UDP-acetyl glycans indicative of HBP activity. These data demonstrate a novel effect of asparagine on cellular metabolism in FH-deficient cells that could be exploited therapeutically.
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Restless legs syndrome (RLS) is characterized by an urge to move legs, usually accompanied by uncomfortable sensations. RLS symptoms generally happen at night and can be relieved by movements. Genetic studies have linked polymorphisms in BTBD9 to a higher risk of RLS. Knockout of BTBD9 homolog in mice (Btbd9) and fly results in RLS-like phenotypes. A dysfunctional dopaminergic system is associated with RLS. However, the function of BTBD9 in the dopaminergic system and RLS is not clear. Here, we made use of the simple Caenorhabditis elegans nervous system. Loss of hpo-9, the worm homolog of BTBD9, resulted in hyperactive egg-laying behavior. Analysis of genetic interactions between hpo-9 and genes for dopamine receptors (dop-1, dop-3) indicated that hpo-9 and dop-1 worked similarly. Reporter assays of dop-1 and dop-3 revealed that hpo-9 knockout led to a significant increase of DOP-3 expression. This appears to be evolutionarily conserved in mice with an increased D2 receptor (D2R) mRNA in the striatum of the Btbd9 knockout mice. Furthermore, the striatal D2R protein was significantly decreased and Dynamin I was increased. Overall, activities of DA neurons in the substantia nigra were not altered, but the peripheral D1R pathway was potentiated in the Btbd9 knockout mice. Finally, we generated and characterized the dopamine neuron-specific Btbd9 knockout mice and detected an active-phase sleepiness, suggesting that dopamine neuron-specific loss of Btbd9 is sufficient to disturb the sleep. Our results suggest that increased activities in the D1R pathway, decreased activities in the D2R pathway, or both may contribute to RLS.
Assuntos
Domínio BTB-POZ/fisiologia , Proteínas de Caenorhabditis elegans/fisiologia , Dopamina/fisiologia , Proteínas do Tecido Nervoso/fisiologia , Síndrome das Pernas Inquietas/fisiopatologia , Animais , Caenorhabditis elegans , Modelos Animais de Doenças , Dopamina/administração & dosagem , Neurônios Dopaminérgicos/fisiologia , Dinamina I/metabolismo , Feminino , Masculino , Camundongos Knockout , Proteínas do Tecido Nervoso/genética , Receptores de Dopamina D2/metabolismo , Substância Negra/fisiologiaRESUMO
Renal medullary carcinoma (RMC) is a rare, aggressive disease that predominantly afflicts individuals of African or Mediterranean descent with sickle cell trait. RMC comprises 1% of all renal cell carcinoma diagnoses with a median overall survival of 13 months. Patients are typically young (median age-22) and male (male:female ratio of 2:1) and tumors are characterized by complete loss of expression of the SMARCB1 tumor suppressor protein. Due to the low incidence of RMC and the disease's aggressiveness, treatment decisions are often based on case reports. Thus, it is critical to develop preclinical models of RMC to better understand the pathogenesis of this disease and to identify effective forms of therapy. Two novel cell line models, UOK353 and UOK360, were derived from primary RMCs that both demonstrated the characteristic SMARCB1 loss. Both cell lines overexpressed EZH2 and other members of the polycomb repressive complex and EZH2 inhibition in RMC tumor spheroids resulted in decreased viability. High throughput drug screening of both cell lines revealed several additional candidate compounds, including bortezomib that had both in vitro and in vivo antitumor activity. The activity of bortezomib was shown to be partially dependent on increased oxidative stress as addition of the N-acetyl cysteine antioxidant reduced the effect on cell proliferation. Combining bortezomib and cisplatin further decreased cell viability both in vitro and in vivo that single agent bortezomib treatment. The UOK353 and UOK360 cell lines represent novel preclinical models for the development of effective forms of therapy for RMC patients.
Assuntos
Carcinoma Medular/patologia , Neoplasias Renais/patologia , Cultura Primária de Células/métodos , Ensaios Antitumorais Modelo de Xenoenxerto/métodos , Animais , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Bortezomib/farmacologia , Bortezomib/uso terapêutico , Carcinoma Medular/tratamento farmacológico , Carcinoma Medular/genética , Autenticação de Linhagem Celular/métodos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Cisplatino/farmacologia , Cisplatino/uso terapêutico , Proteína Potenciadora do Homólogo 2 de Zeste/genética , Proteína Potenciadora do Homólogo 2 de Zeste/metabolismo , Humanos , Neoplasias Renais/tratamento farmacológico , Neoplasias Renais/genética , Camundongos , Camundongos Nus , Proteína SMARCB1/genética , Proteína SMARCB1/metabolismo , Células Tumorais CultivadasRESUMO
As a class of peptide hormone, plant natriuretic peptides (PNPs) play an important role in maintaining water and salt balance in plants, as well as in the physiological processes of biotic stress and pathogen resistance. However, in plants, except for some PNPs, such as the Arabidopsis thaliana PNP-A (AtPNP-A), of which the function has not yet been thoroughly revealed, few PNPs in other plants have been reported. In this study, a PNP-A (ScPNP-A) has been identified and characterized in Stellera chamaejasme for the first time. ScPNP-A is a double-psi beta-barrel (DPBB) fold containing protein and is localized in the extracellular (secreted) space. In S. chamaejasme, the expression of ScPNP-A was significantly up-regulated by salt, drought and cold stress. Changes at the physiological and biochemical levels and the expression of resistance-related genes indicated that overexpression of ScPNP-A can significantly improve salt, drought and freezing tolerance in Arabidopsis. ScPNP-A could stimulate the opening, not the closing of stomata, and its expression was not enhanced by external application of ABA. Furthermore, overexpression of ScPNP-A resulted in the elevated expression of genes in the ABA biosynthesis and reception pathway. These suggested that there may be some cross-talk between ScPNP-A and the ABA-dependent signaling pathways to regulate water related stress, however further experimentation is required to understand this relationship. In addition, overexpression of ScPNP-A can enhance the resistance to pathogens by enhancing SAR in Arabidopsis. These results indicate that ScPNP-A could function as a positive regulator in plant response to biotic stress and abiotic stress.
Assuntos
Arabidopsis , Peptídeos Natriuréticos , Estresse Fisiológico , Arabidopsis/genética , Arabidopsis/metabolismo , Secas , Expressão Gênica , Peptídeos Natriuréticos/genética , Peptídeos Natriuréticos/metabolismo , Estômatos de Plantas/genética , Estômatos de Plantas/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Estresse Fisiológico/genética , Thymelaeaceae/genéticaRESUMO
Restless legs syndrome (RLS) is a neurological disorder characterized by an urge to move and uncomfortable sensations. Genetic studies have identified polymorphisms in up to 19 risk loci, including MEIS1 and BTBD9. Rodents deficient in either homolog show RLS-like phenotypes. However, whether MEIS1 and BTBD9 interact in vivo is unclear. Here, with C. elegans, we observed that the hyperactive egg-laying behavior caused by loss of BTBD9 homolog was counteracted by knockdown of MEIS1 homolog. This was further investigated in mutant mice with Btbd9, Meis1, or both knocked out. The double knockout mice showed an earlier onset of the motor deficit in the wheel running test but did not have increased sensitivity to the heat stimuli as observed in single KOs. Meis1 protein level was not influenced by Btbd9 deficiency, and Btbd9 transcription was not affected by Meis1 haploinsufficiency. Our results demonstrate that MEIS1 and BTBD9 do not regulate each other.