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1.
Food Chem ; 448: 139059, 2024 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-38531295

RESUMO

Plant-based (PB) food products have surged in popularity over the past decade. Available PB products in the UK market were extracted from NielsenIQ Brandbank and compared with animal-based (AB) counterparts in their nutrient contents and calculated Nutri-Scores. The amino acid contents of four beef products and their PB alternatives were analysed by LC-MS/MS. PB products consistently exhibited significantly higher fibre content across all food groups. Protein was significantly higher in AB products from all food groups except beef and ready meals. PB products were more likely to have higher Nutri-Scores compared to AB counterparts, albeit with greater score variability within each food group. Nutrient fortifications were primarily focused on dairy and ready meals; the most supplemented nutrient was vitamin B12 (found in 15% of all products). A higher proportion of EAAs in relation to total protein content was observed in all beef products.


Assuntos
Aminoácidos , Suplementos Nutricionais , Valor Nutritivo , Animais , Aminoácidos/análise , Reino Unido , Bovinos , Suplementos Nutricionais/análise , Espectrometria de Massas em Tandem
2.
Redox Biol ; 67: 102878, 2023 11.
Artigo em Inglês | MEDLINE | ID: mdl-37703668

RESUMO

Cruciferous-rich diets, particularly broccoli, have been associated with reduced risk of developing cancers of various sites, cardiovascular disease and type-2 diabetes. Sulforaphane (SF), a sulfur-containing broccoli-derived metabolite, has been identified as the major bioactive compound mediating these health benefits. Sulforaphane is a potent dietary activator of the transcription factor Nuclear factor erythroid-like 2 (NRF2), the master regulator of antioxidant cell capacity responsible for inducing cytoprotective genes, but its role in glucose homeostasis remains unclear. In this study, we set to test the hypothesis that SF regulates glucose metabolism and ameliorates glucose overload and its resulting oxidative stress by inducing NRF2 in human hepatoma HepG2 cells. HepG2 cells were exposed to varying glucose concentrations: basal (5.5 mM) and high glucose (25 mM), in the presence of physiological concentrations of SF (10 µM). SF upregulated the expression of glutathione (GSH) biosynthetic genes and significantly increased levels of reduced GSH. Labelled glucose and glutamine experiments to measure metabolic fluxes identified that SF increased intracellular utilisation of glycine and glutamate by redirecting the latter away from the TCA cycle and increased the import of cysteine from the media, likely to support glutathione synthesis. Furthermore, SF altered pathways generating NADPH, the necessary cofactor for oxidoreductase reactions, namely pentose phosphate pathway and 1C-metabolism, leading to the redirection of glucose away from glycolysis and towards PPP and of methionine towards methylation substrates. Finally, transcriptomic and targeted metabolomics LC-MS analysis of NRF2-KD HepG2 cells generated using CRISPR-Cas9 genome editing revealed that the above metabolic effects are mediated through NRF2. These results suggest that the antioxidant properties of cruciferous diets are intricately connected to their metabolic benefits.


Assuntos
Antioxidantes , Fator 2 Relacionado a NF-E2 , Humanos , Antioxidantes/farmacologia , Antioxidantes/metabolismo , Fator 2 Relacionado a NF-E2/genética , Fator 2 Relacionado a NF-E2/metabolismo , Isotiocianatos/farmacologia , Estresse Oxidativo , Glutationa/metabolismo , Homeostase , Glucose
3.
Nutrients ; 14(16)2022 Aug 10.
Artigo em Inglês | MEDLINE | ID: mdl-36014767

RESUMO

Diets rich in cruciferous vegetables have been associated with a lower risk of incidence and progression of prostate cancer. Sulforaphane, an isothiocyanate derived from 4-methylsulphinylbutyl glucosinolate (glucoraphanin) that accumulates in certain of these vegetables, notably broccoli, has been implicated in their protective effects. Likewise, the consumption of garlic and its sulphur-containing compounds such as alliin have been associated with a reduction in risk of prostate cancer. In this study, we tested whether consuming glucoraphanin derived from broccoli seeds and alliin derived from garlic resulted in the occurrence of these potential bioactive compounds in the prostate, which may contribute to our understanding of the putative protective effects of these dietary components. We recruited 42 men scheduled for a trans-perineal prostate biopsy into a randomised, double-blinded, 2 × 2-factorial dietary supplement four-week intervention study, and 39 completed the study. The two active interventions were supplements providing glucoraphanin from broccoli (BroccoMax®) and alliin from garlic (Kwai Heartcare®). Following the intervention, prostate biopsy tissue was analysed for the presence of sulforaphane and its thiol conjugates and for alliin and associated metabolites. Sulforaphane occurred in significantly higher levels in the prostate tissue (both within the transition and peripheral zone) of men consuming the glucoraphanin containing supplements (p < 0.0001) compared to men not consuming these supplements. However, while alliin and alliin-derived metabolites were detected within the prostate, there was no significant difference in the concentrations of these compounds in the prostate of men consuming supplements derived from garlic compared to men not consuming these supplements.


Assuntos
Allium , Brassica , Neoplasias da Próstata , Antioxidantes/metabolismo , Brassica/metabolismo , Cisteína/análogos & derivados , Glucosinolatos/metabolismo , Humanos , Imidoésteres/metabolismo , Isotiocianatos/metabolismo , Masculino , Próstata/metabolismo , Neoplasias da Próstata/metabolismo , Neoplasias da Próstata/prevenção & controle , Sulfóxidos
4.
Cell Rep ; 40(3): 111130, 2022 07 19.
Artigo em Inglês | MEDLINE | ID: mdl-35858549

RESUMO

Peripheral nervous system (PNS) injuries initiate transcriptional changes in glial cells and sensory neurons that promote axonal regeneration. While the factors that initiate the transcriptional changes in glial cells are well characterized, the full range of stimuli that initiate the response of sensory neurons remain elusive. Here, using a genetic model of glial cell ablation, we find that glial cell loss results in transient PNS demyelination without overt axonal loss. By profiling sensory ganglia at single-cell resolution, we show that glial cell loss induces a transcriptional injury response preferentially in proprioceptive and Aß RA-LTMR neurons. The transcriptional response of sensory neurons to mechanical injury has been assumed to be a cell-autonomous response. By identifying a similar response in non-injured, demyelinated neurons, our study suggests that this represents a non-cell-autonomous transcriptional response of sensory neurons to glial cell loss and demyelination.


Assuntos
Doenças Desmielinizantes , Neuroglia , Humanos , Neuroglia/fisiologia , Sistema Nervoso Periférico , Células Receptoras Sensoriais
5.
Front Plant Sci ; 13: 855707, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35432397

RESUMO

Broccoli cultivars that have enhanced accumulation of methionine-derived glucosinolates have been developed through the introgression of a novel allele of the MYB28 transcription factor from the wild species Brassica villosa. Through a novel k-mer approach, we characterised the extent of the introgression of unique B. villosa genome sequences into high glucosinolate broccoli genotypes. RNAseq analyses indicated that the introgression of the B. villosa MYB28 C2 allele resulted in the enhanced expression of the MYB28 transcription factor, and modified expression of genes associated with sulphate absorption and reduction, and methionine and glucosinolate biosynthesis when compared to standard broccoli. A adenine-thymine (AT) short tandem repeat (STR) was identified within the 5' untranslated region (UTR) B. villosa MYB28 allele that was absent from two divergent cultivated forms of Brassica oleracea, which may underpin the enhanced expression of B. villosa MYB28.

6.
Eur Urol Oncol ; 5(4): 412-419, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35450835

RESUMO

BACKGROUND: Bacteria play a suspected role in the development of several cancer types, and associations between the presence of particular bacteria and prostate cancer have been reported. OBJECTIVE: To provide improved characterisation of the prostate and urine microbiome and to investigate the prognostic potential of the bacteria present. DESIGN, SETTING, AND PARTICIPANTS: Microbiome profiles were interrogated in sample collections of patient urine (sediment microscopy: n = 318, 16S ribosomal amplicon sequencing: n = 46; and extracellular vesicle RNA-seq: n = 40) and cancer tissue (n = 204). OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: Microbiomes were assessed using anaerobic culture, population-level 16S analysis, RNA-seq, and whole genome DNA sequencing. RESULTS AND LIMITATIONS: We demonstrate an association between the presence of bacteria in urine sediments and higher D'Amico risk prostate cancer (discovery, n = 215 patients, p < 0.001; validation, n = 103, p < 0.001, χ2 test for trend). Characterisation of the bacterial community led to the (1) identification of four novel bacteria (Porphyromonas sp. nov., Varibaculum sp. nov., Peptoniphilus sp. nov., and Fenollaria sp. nov.) that were frequently found in patient urine, and (2) definition of a patient subgroup associated with metastasis development (p = 0.015, log-rank test). The presence of five specific anaerobic genera, which includes three of the novel isolates, was associated with cancer risk group, in urine sediment (p = 0.045, log-rank test), urine extracellular vesicles (p = 0.039), and cancer tissue (p = 0.035), with a meta-analysis hazard ratio for disease progression of 2.60 (95% confidence interval: 1.39-4.85; p = 0.003; Cox regression). A limitation is that functional links to cancer development are not yet established. CONCLUSIONS: This study characterises prostate and urine microbiomes, and indicates that specific anaerobic bacteria genera have prognostic potential. PATIENT SUMMARY: In this study, we investigated the presence of bacteria in patient urine and the prostate. We identified four novel bacteria and suggest a potential prognostic utility for the microbiome in prostate cancer.


Assuntos
Microbiota , Neoplasias da Próstata , Bactérias/genética , Humanos , Masculino , Microbiota/genética , Próstata/patologia , Neoplasias da Próstata/patologia , RNA Ribossômico 16S/genética
7.
Calcif Tissue Int ; 110(3): 273-284, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-34870723

RESUMO

The human microbiota functions at the interface between diet, medication-use, lifestyle, host immune development and health. It is therefore closely aligned with many of the recognised modifiable factors that influence bone mass accrual in the young, and bone maintenance and skeletal decline in older populations. While understanding of the relationship between micro-organisms and bone health is still in its infancy, two decades of broader microbiome research and discovery supports a role of the human gut microbiome in the regulation of bone metabolism and pathogenesis of osteoporosis as well as its prevention and treatment. Pre-clinical research has demonstrated biological interactions between the microbiome and bone metabolism. Furthermore, observational studies and randomized clinical trials have indicated that therapeutic manipulation of the microbiota by oral administration of probiotics may influence bone turnover and prevent bone loss in humans. In this paper, we summarize the content, discussion and conclusions of a workshop held by the Osteoporosis and Bone Research Academy of the Royal Osteoporosis Society in October, 2020. We provide a detailed review of the literature examining the relationship between the microbiota and bone health in animal models and in humans, as well as formulating the agenda for key research priorities required to advance this field. We also underscore the potential pitfalls in this research field that should be avoided and provide methodological recommendations to facilitate bridging the gap from promising concept to a potential cause and intervention target for osteoporosis.


Assuntos
Microbioma Gastrointestinal , Microbiota , Osteoporose , Probióticos , Animais , Osso e Ossos/metabolismo , Microbioma Gastrointestinal/fisiologia , Osteoporose/metabolismo , Osteoporose/prevenção & controle , Probióticos/uso terapêutico
8.
CRISPR J ; 4(3): 416-426, 2021 06.
Artigo em Inglês | MEDLINE | ID: mdl-34152214

RESUMO

Discoveries in model plants grown under optimal conditions can provide important directions for crop improvement. However, it is important to verify whether results can be translated to crop plants grown in the field. In this study, we sought to study the role of MYB28 in the regulation of aliphatic glucosinolate (A-GSL) biosynthesis and associated sulfur metabolism in field-grown Brassica oleracea with the use of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas9 gene-editing technology. We describe the first myb28 knockout mutant in B. oleracea, and the first CRISPR field trial in the United Kingdom approved and regulated by the UK Department for Environment, Food & Rural Affairs after the reclassification of gene-edited crops as genetically modified organisms by the European Court of Justice on July 25, 2018. We report that knocking out myb28 results in downregulation of A-GSL biosynthesis genes and reduction in accumulation of the methionine-derived glucosinolate, glucoraphanin, in leaves and florets of field-grown myb28 mutant broccoli plants, whereas accumulation of sulfate, S-methyl cysteine sulfoxide, and indole glucosinolate in leaf and floret tissues remained unchanged. These results demonstrate the potential of gene-editing approaches to translate discoveries in fundamental biological processes for improved crop performance.


Assuntos
Brassica/genética , Brassica/metabolismo , Sistemas CRISPR-Cas , Edição de Genes/métodos , Glucosinolatos/biossíntese , Glucosinolatos/genética , Histona Acetiltransferases/genética , Histona Acetiltransferases/metabolismo , Proteínas de Arabidopsis , Produtos Agrícolas/genética , Produtos Agrícolas/metabolismo , Expressão Gênica , Oximas , Plantas Geneticamente Modificadas , Sulfóxidos/metabolismo , Reino Unido
9.
Physiol Rep ; 8(13): e14482, 2020 07.
Artigo em Inglês | MEDLINE | ID: mdl-32643289

RESUMO

BACKGROUND AND AIMS: Nonalcoholic fatty liver disease (NAFLD) begins with steatosis, where a mixed macrovesicular pattern of large and small lipid droplets (LDs) develops. Since in vitro models recapitulating this are limited, the aims of this study were to develop mixed macrovesicular steatosis in immortalized hepatocytes and investigate effects on intracellular metabolism by altering nutritional substrates. METHODS: Huh7 cells were cultured in 11 mM glucose and 2% human serum (HS) for 7 days before additional sugars and fatty acids (FAs), either with 200 µM FAs (low fat low sugar; LFLS), 5.5 mM fructose + 200 µM FAs (low fat high sugar; LFHS), or 5.5 mM fructose + 800 µM FAs (high fat high sugar; HFHS), were added for 7 days. FA metabolism, lipid droplet characteristics, and transcriptomic signatures were investigated. RESULTS: Between the LFLS and LFHS conditions, there were few notable differences. In the HFHS condition, intracellular triacylglycerol (TAG) was increased and the LD pattern and distribution was similar to that found in primary steatotic hepatocytes. HFHS-treated cells had lower levels of de novo-derived FAs and secreted larger, TAG-rich lipoprotein particles. RNA sequencing and gene set enrichment analysis showed changes in several pathways including those involved in metabolism and cell cycle. CONCLUSIONS: Repeated doses of HFHS treatment resulted in a cellular model of NAFLD with a mixed macrovesicular LD pattern and metabolic dysfunction. Since these nutrients have been implicated in the development of NAFLD in humans, the model provides a good physiological basis for studying NAFLD development or regression in vitro.


Assuntos
Ácidos Graxos/metabolismo , Glucose/metabolismo , Hepatócitos/metabolismo , Gotículas Lipídicas/metabolismo , Hepatopatia Gordurosa não Alcoólica/metabolismo , Linhagem Celular Tumoral , Células Cultivadas , Hepatócitos/patologia , Humanos , Gotículas Lipídicas/patologia , Hepatopatia Gordurosa não Alcoólica/genética , Transcriptoma
10.
Nutrients ; 11(9)2019 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-31540470

RESUMO

Prostate cancer has become the most common form of non-cutaneous (internal) malignancy in men, accounting for 26% of all new male visceral cancer cases in the UK. The aetiology and pathogenesis of prostate cancer are not understood, but given the age-adjusted geographical variations in prostate cancer incidence quoted in epidemiological studies, there is increasing interest in nutrition as a relevant factor. In particular, foods rich in phytochemicals have been proposed to reduce the risk of prostate cancer. Epidemiological studies have reported evidence that plant-based foods including cruciferous vegetables, garlic, tomatoes, pomegranate and green tea are associated with a significant reduction in the progression of prostate cancer. However, while there is well-documented mechanistic evidence at a cellular level of the manner by which individual dietary components may reduce the risk of prostate cancer or its progression, evidence from intervention studies is limited. Moreover, clinical trials investigating the link between the dietary bioactives found in these foods and prostate cancer have reported varied conclusions. Herein, we review the plant bioactives for which there is substantial evidence from epidemiological and human intervention studies. The aim of this review is to provide important insights into how particular plant bioactives (e.g., sulphur-containing compounds, carotenoids and polyphenols) present in commonly consumed food groups may influence the development and progression of prostate cancer.


Assuntos
Compostos Fitoquímicos , Extratos Vegetais , Neoplasias da Próstata , Ensaios Clínicos como Assunto , Humanos , Masculino , Neoplasias da Próstata/dietoterapia , Neoplasias da Próstata/tratamento farmacológico , Neoplasias da Próstata/prevenção & controle , Verduras
11.
Mol Nutr Food Res ; 63(20): e1900461, 2019 10.
Artigo em Inglês | MEDLINE | ID: mdl-31410992

RESUMO

SCOPE: Observational studies have associated consumption of cruciferous vegetables with reduced risk of prostate cancer. This effect has been associated with the degradation products of glucosinolates-thioglycosides that accumulate within crucifers. The possible role of S-methyl cysteine sulfoxide, a metabolite that also accumulates in cruciferous vegetables, and its derivatives, in cancer prevention is relatively unexplored compared to glucosinolate derivatives. The hypothesis that consuming a broccoli soup results in the accumulation of sulfate (a SMCSO derivative) and other broccoli-derived metabolites in prostate tissue is tested. METHODS AND RESULTS: Eighteen men scheduled for transperineal prostate biopsy were recruited into a 4-week parallel single blinded diet supplementation study (NCT02821728). Nine men supplemented their diet with three 300 mL portions of a broccoli soup each week for four weeks prior to surgery. Analyses of prostate biopsy tissues reveal no detectable levels of glucosinolates and derivatives. In contrast, SMCSO is detected in prostate tissues of the participants, with significantly higher levels in tissue of men in the supplementation arm. SMCSO was also found in blood and urine samples from a previous intervention study with the identical broccoli soup. CONCLUSION: The consequences of SMCSO accumulation in prostate tissues and its potential role in prevention of prostate cancer remains to be investigated.


Assuntos
Brassica , Próstata/metabolismo , Sulfóxidos/metabolismo , Idoso , Allium , Suplementos Nutricionais , Glucosinolatos/metabolismo , Humanos , Imidoésteres/metabolismo , Isotiocianatos/metabolismo , Masculino , Pessoa de Meia-Idade , Oximas , Neoplasias da Próstata/patologia , Neoplasias da Próstata/prevenção & controle , Método Simples-Cego
12.
Am J Clin Nutr ; 109(4): 1133-1144, 2019 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-30982861

RESUMO

BACKGROUND: Epidemiological evidence suggests that consumption of cruciferous vegetables is associated with reduced risk of prostate cancer progression, largely attributed to the biological activity of glucosinolate degradation products, such as sulforaphane derived from glucoraphanin. Because there are few therapeutic interventions for men on active surveillance for prostate cancer to reduce the risk of cancer progression, dietary approaches are an appealing option for patients. OBJECTIVE: We evaluated whether consumption of a glucoraphanin-rich broccoli soup for 1 y leads to changes in gene expression in prostate tissue of men with localized prostate cancer. METHODS: Forty-nine men on active surveillance completed a 3-arm parallel randomized double-blinded intervention study for 12 mo and underwent transperineal template biopsy procedures and dietary assessment at the start and end of the study. Patients received a weekly 300 mL portion of soup made from a standard broccoli (control) or from 1 of 2 experimental broccoli genotypes with enhanced concentrations of glucoraphanin, delivering 3 and 7 times that of the control, respectively. Gene expression in tissues from each patient obtained before and after the dietary intervention was quantified by RNA sequencing followed by gene set enrichment analyses. RESULTS: In the control arm, there were several hundred changes in gene expression in nonneoplastic tissue during the 12 mo. These were associated with an increase in expression of potentially oncogenic pathways including inflammation processes and epithelial-mesenchymal transition. Changes in gene expression and associated oncogenic pathways were attenuated in men on the glucoraphanin-rich broccoli soup in a dose-dependent manner. Although the study was not powered to assess clinical progression, an inverse association between consumption of cruciferous vegetables and cancer progression was observed. CONCLUSION: Consuming glucoraphanin-rich broccoli soup affected gene expression in the prostate of men on active surveillance, consistent with a reduction in the risk of cancer progression. This trial was registered at clinicaltrials.gov as NCT01950143.


Assuntos
Brassica/metabolismo , Glucosinolatos/metabolismo , Imidoésteres/metabolismo , Isotiocianatos/metabolismo , Próstata/metabolismo , Neoplasias da Próstata/genética , Neoplasias da Próstata/prevenção & controle , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Oximas , Neoplasias da Próstata/metabolismo , Sulfóxidos , Transcrição Gênica , Adulto Jovem
13.
Methods Mol Biol ; 1936: 295-310, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30820906

RESUMO

Genetic mouse models facilitate investigation of mechanisms underpinning human diseases and aid the development of novel therapeutic treatments. To better understand the demyelination and remyelination processes in adult-onset demyelinating diseases like multiple sclerosis (MS), we have developed the DTA mouse model system that allows for the widespread ablation of the mature oligodendrocytes, resulting in demyelination throughout the central nervous system (CNS). Induction of oligodendrocyte death in young adult DTA mice causes extensive CNS demyelination that leads to a severe neurological disease, followed by a full recovery that is associated with extensive replenishment of oligodendrocytes and remyelination. Thus, the DTA mouse enables investigation of the mechanisms that promote remyelination in MS and other adult-onset demyelinating diseases. Approximately 30 weeks later, the recovered DTA mice develop a fatal secondary demyelinating disease that is mediated by autoimmune T cells. Therefore, the DTA mouse model is also ideal for elucidating the role of oligodendrocyte death in eliciting autoimmunity in MS. In this chapter we describe the methods we used to generate the DTA mouse model and to analyze both the primary and secondary demyelinating diseases in DTA mice.


Assuntos
Doenças Desmielinizantes/imunologia , Toxina Diftérica/genética , Modelos Animais de Doenças , Fragmentos de Peptídeos/genética , Animais , Sistema Nervoso Central/citologia , Sistema Nervoso Central/imunologia , Doenças Desmielinizantes/genética , Feminino , Humanos , Masculino , Camundongos , Microscopia Eletrônica de Transmissão , Oligodendroglia/citologia , Oligodendroglia/imunologia , Oligodendroglia/patologia , Remielinização
14.
FASEB J ; 33(2): 2372-2387, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30277819

RESUMO

NF-E2-related factor 2 (NRF2) transcription factor has a fundamental role in cell homeostasis maintenance as one of the master regulators of oxidative and electrophilic stress responses. Previous studies have shown that a regulatory connection exists between NRF2 and autophagy during reactive oxygen species-generated oxidative stress. The aim of the present study was to investigate how autophagy is turned off during prolonged oxidative stress, to avoid overeating and destruction of essential cellular components. AMPK is a key cellular energy sensor highly conserved in eukaryotic organisms, and it has an essential role in autophagy activation at various stress events. Here the role of human AMPK and its Caenorhabditis elegans counterpart AAK-2 was explored upon oxidative stress. We investigated the regulatory connection between NRF2 and AMPK during oxidative stress induced by tert-butyl hydroperoxide (TBHP) in HEK293T cells and C. elegans. Putative conserved NRF2/protein skinhead-1 binding sites were found in AMPK/aak-2 genes by in silico analysis and were later confirmed experimentally by using EMSA. After addition of TBHP, NRF2 and AMPK showed a quick activation; AMPK was later down-regulated, however, while NRF2 level remained high. Autophagosome formation and Unc-51-like autophagy activating kinase 1 phosphorylation were initially stimulated, but they returned to basal values after 4 h of TBHP treatment. The silencing of NRF2 resulted in a constant activation of AMPK leading to hyperactivation of autophagy during oxidative stress. We observed the same effects in C. elegans demonstrating the conservation of this self-defense mechanism to save cells from hyperactivated autophagy upon prolonged oxidative stress. We conclude that NRF2 negatively regulates autophagy through delayed down-regulation of the expression of AMPK upon prolonged oxidative stress. This regulatory connection between NRF2 and AMPK may have an important role in understanding how autophagy is regulated in chronic human morbidities characterized by oxidative stress, such as neurodegenerative diseases, certain cancer types, and in metabolic diseases.-Kosztelnik, M., Kurucz, A., Papp, D., Jones, E., Sigmond, T., Barna, J., Traka, M. H., Lorincz, T., Szarka, A., Banhegyi, G., Vellai, T., Korcsmaros, T., Kapuy, O. Suppression of AMPK/aak-2 by NRF2/SKN-1 down-regulates autophagy during prolonged oxidative stress.


Assuntos
Autofagia , Proteínas de Caenorhabditis elegans/antagonistas & inibidores , Proteínas de Caenorhabditis elegans/metabolismo , Proteínas de Ligação a DNA/metabolismo , Regulação da Expressão Gênica , Fator 2 Relacionado a NF-E2/metabolismo , Estresse Oxidativo , Proteínas Quinases/química , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Fatores de Transcrição/metabolismo , Quinases Proteína-Quinases Ativadas por AMP , Proteínas Quinases Ativadas por AMP , Animais , Caenorhabditis elegans/genética , Caenorhabditis elegans/crescimento & desenvolvimento , Caenorhabditis elegans/metabolismo , Proteínas de Caenorhabditis elegans/genética , Proteínas de Ligação a DNA/genética , Regulação para Baixo , Células HEK293 , Humanos , Fator 2 Relacionado a NF-E2/genética , Oxirredução , Fosforilação , Proteínas Quinases/genética , Proteínas Quinases/metabolismo , Proteínas Serina-Treonina Quinases/genética , Proteínas Serina-Treonina Quinases/metabolismo , Fatores de Transcrição/genética
15.
Mol Nutr Food Res ; 62(18): e1700911, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-29266773

RESUMO

SCOPE: Broccoli accumulates 4-methylsulphinylbutyl glucosinolate (glucoraphanin) which is hydrolyzed to the isothiocyanate sulforaphane. Through the introgression of novel alleles of the Myb28 transcription factor from Brassica villosa, broccoli genotypes have been developed that have enhanced levels of glucoraphanin. This study seeks to quantify the exposure of human tissues to glucoraphanin and sulforaphane following consumption of broccoli with contrasting Myb28 genotypes. METHODS AND RESULTS: Ten participants are recruited into a three-phase, double-blinded, randomized crossover trial (NCT02300324), with each phase comprising consumption of 300 g of a soup made from broccoli of one of three Myb28 genotypes (Myb28B/B , Myb28B/V , Myb28V/V ). Plant myrosinases are intentionally denatured during soup manufacture. Threefold and fivefold higher levels of sulforaphane occur in the circulation following consumption of Myb28V/B and Myb28V/V broccoli soups, respectively. The percentage of sulforaphane excreted in 24 h relative to the amount of glucoraphanin consumed varies among volunteers from 2 to 15%, but does not depend on the broccoli genotype. CONCLUSION: This is the first study to report the bioavailability of glucoraphanin and sulforaphane from soups made with novel broccoli varieties. The presence of one or two Myb28V alleles results in enhanced delivery of sulforaphane to the systemic circulation.


Assuntos
Brassica/química , Glucosinolatos/farmacocinética , Imidoésteres/farmacocinética , Isotiocianatos/farmacocinética , Adolescente , Adulto , Idoso , Alelos , Disponibilidade Biológica , Brassica/genética , Estudos Cross-Over , Dieta , Método Duplo-Cego , Feminino , Genótipo , Técnicas de Genotipagem , Glucosinolatos/sangue , Glucosinolatos/urina , Glicosídeo Hidrolases/genética , Glicosídeo Hidrolases/metabolismo , Humanos , Isotiocianatos/sangue , Isotiocianatos/urina , Masculino , Pessoa de Meia-Idade , Oximas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Sulfóxidos , Espectrometria de Massas em Tandem , Adulto Jovem
16.
Oncotarget ; 8(49): 84902-84916, 2017 Oct 17.
Artigo em Inglês | MEDLINE | ID: mdl-29156692

RESUMO

The human prostate gland comprises three distinct anatomical glandular zones, namely the peripheral, central and transitional zones. Although prostate cancer can arise throughout the prostate, it is more frequent in the peripheral zone. In contrast, hyperplasia occurs most frequently in the transitional zone. In this paper, we test the hypothesis that peripheral and transitional zones have distinct metabolic adaptations that may underlie their different inherent predispositions to cancer and hyperplasia. In order to do this, we undertook RNA sequencing and high-throughput metabolic analyses of non-cancerous tissue from the peripheral and transitional zones of patients undergoing prostatectomy. Integrated analysis of RNAseq and metabolomic data revealed that transcription of genes involved in lipid biosynthesis is higher in the peripheral zone, which was mirrored by an increase in fatty acid metabolites, such as lysolipids. The peripheral zone also exhibited increased fatty acid catabolic activity and contained higher level of neurotransmitters. Such increased capacity for de novo lipogenesis and fatty acid oxidation, which is characteristic of prostate cancer, can potentially provide a permissive growth environment within the peripheral zone for cancer growth and also transmit a metabolic growth advantage to newly emerging clones themselves. This lipo-rich priming may explain the observed susceptibility of the peripheral zone to oncogenesis.

17.
Sci Rep ; 7(1): 3398, 2017 06 13.
Artigo em Inglês | MEDLINE | ID: mdl-28611391

RESUMO

Osteoarthritis is a major cause of disability and there is no current pharmaceutical treatment which can prevent the disease or slow its progression. Dietary advice or supplementation is clearly an attractive option since it has low toxicity and ease of implementation on a population level. We have previously demonstrated that sulforaphane, a dietary isothiocyanate derived from its glucosinolate precursor which is found in broccoli, can prevent cartilage destruction in cells, in in vitro and in vivo models of osteoarthritis. As the next phase of this research, we enrolled 40 patients with knee osteoarthritis undergoing total knee replacement into a proof-of-principle trial. Patients were randomised to either a low or high glucosinolate diet for 14 days prior to surgery. We detected ITCs in the synovial fluid of the high glucosinolate group, but not the low glucosinolate group. This was mirrored by an increase in ITCs and specifically sulforaphane in the plasma. Proteomic analysis of synovial fluid showed significantly distinct profiles between groups with 125 differentially expressed proteins. The functional consequence of this diet will now be tested in a clinical trial.


Assuntos
Biomarcadores/metabolismo , Brassica/efeitos adversos , Isotiocianatos/metabolismo , Articulação do Joelho/fisiopatologia , Osteoartrite do Joelho/etiologia , Líquido Sinovial/metabolismo , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Osteoartrite do Joelho/metabolismo , Osteoartrite do Joelho/patologia , Proteômica
18.
Prostate ; 76(14): 1326-37, 2016 10.
Artigo em Inglês | MEDLINE | ID: mdl-27403764

RESUMO

BACKGROUND: Acylcarnitines are intermediates of fatty acid oxidation and accumulate as a consequence of the metabolic dysfunction resulting from the insufficient integration between ß-oxidation and the tricarboxylic acid (TCA) cycle. The aim of this study was to investigate whether acylcarnitines accumulate in prostate cancer tissue, and whether their biological actions could be similar to those of dihydrotestosterone (DHT), a structurally related compound associated with cancer development. METHODS: Levels of palmitoylcarnitine (palcar), a C16:00 acylcarnitine, were measured in prostate tissue using LC-MS/MS. The effect of palcar on inflammatory cytokines and calcium (Ca(2+) ) influx was investigated in in vitro models of prostate cancer. RESULTS: We observed a significantly higher level of palcar in prostate cancerous tissue compared to benign tissue. High levels of palcar have been associated with increased gene expression and secretion of the pro-inflammatory cytokine IL-6 in cancerous PC3 cells, compared to normal PNT1A cells. Furthermore, we found that high levels of palcar induced a rapid Ca(2+) influx in PC3 cells, but not in DU145, BPH-1, or PNT1A cells. This pattern of Ca(2+) influx was also observed in response to DHT. Through the use of whole genome arrays we demonstrated that PNT1A cells exposed to palcar or DHT have a similar biological response. CONCLUSIONS: This study suggests that palcar might act as a potential mediator for prostate cancer progression through its effect on (i) pro-inflammatory pathways, (ii) Ca(2+) influx, and (iii) DHT-like effects. Further studies need to be undertaken to explore whether this class of compounds has different biological functions at physiological and pathological levels. Prostate 76:1326-1337, 2016. © 2016 The Authors. The Prostate published by Wiley Periodicals, Inc.


Assuntos
Cálcio/metabolismo , Mediadores da Inflamação/metabolismo , Palmitoilcarnitina/metabolismo , Neoplasias da Próstata/metabolismo , Transdução de Sinais/fisiologia , Sobrevivência Celular/fisiologia , Células Cultivadas , Cromatografia Líquida , Humanos , Masculino , Espectrometria de Massas , Palmitoilcarnitina/análise , Próstata/metabolismo , Neoplasias da Próstata/patologia
19.
Development ; 143(13): 2356-66, 2016 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-27226321

RESUMO

The tumor suppressor protein adenomatous polyposis coli (APC) is multifunctional - it participates in the canonical Wnt/ß-catenin signal transduction pathway as well as modulating cytoskeleton function. Although APC is expressed by Schwann cells, the role that it plays in these cells and in the myelination of the peripheral nervous system (PNS) is unknown. Therefore, we used the Cre-lox approach to generate a mouse model in which APC expression is specifically eliminated from Schwann cells. These mice display hindlimb weakness and impaired axonal conduction in sciatic nerves. Detailed morphological analyses revealed that APC loss delays radial axonal sorting and PNS myelination. Furthermore, APC loss delays Schwann cell differentiation in vivo, which correlates with persistent activation of the Wnt signaling pathway and results in perturbed extension of Schwann cell processes and disrupted lamellipodia formation. In addition, APC-deficient Schwann cells display a transient diminution of proliferative capacity. Our data indicate that APC is required by Schwann cells for their timely differentiation to mature, myelinating cells and plays a crucial role in radial axonal sorting and PNS myelination.


Assuntos
Proteína da Polipose Adenomatosa do Colo/metabolismo , Axônios/metabolismo , Bainha de Mielina/metabolismo , Sistema Nervoso Periférico/metabolismo , Animais , Diferenciação Celular/genética , Membro Posterior/patologia , Integrases/metabolismo , Camundongos , Pseudópodes/metabolismo , Células de Schwann/citologia , Células de Schwann/metabolismo , Nervo Isquiático/metabolismo , Via de Sinalização Wnt/genética
20.
Nat Neurosci ; 19(1): 65-74, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26656646

RESUMO

Although multiple sclerosis is a common neurological disorder, the origin of the autoimmune response against myelin, which is the characteristic feature of the disease, remains unclear. To investigate whether oligodendrocyte death could cause this autoimmune response, we examined the oligodendrocyte ablation Plp1-CreER(T);ROSA26-eGFP-DTA (DTA) mouse model. Approximately 30 weeks after recovering from oligodendrocyte loss and demyelination, DTA mice develop a fatal secondary disease characterized by extensive myelin and axonal loss. Strikingly, late-onset disease was associated with increased numbers of T lymphocytes in the CNS and myelin oligodendrocyte glycoprotein (MOG)-specific T cells in lymphoid organs. Transfer of T cells derived from DTA mice to naive recipients resulted in neurological defects that correlated with CNS white matter inflammation. Furthermore, immune tolerization against MOG ameliorated symptoms. Overall, these data indicate that oligodendrocyte death is sufficient to trigger an adaptive autoimmune response against myelin, suggesting that a similar process can occur in the pathogenesis of multiple sclerosis.


Assuntos
Autoimunidade/imunologia , Morte Celular/imunologia , Doenças Autoimunes Desmielinizantes do Sistema Nervoso Central/imunologia , Bainha de Mielina/imunologia , Glicoproteína Mielina-Oligodendrócito/imunologia , Oligodendroglia , Linfócitos T/imunologia , Animais , Antineoplásicos Hormonais/farmacologia , Contagem de Células , Doenças Autoimunes Desmielinizantes do Sistema Nervoso Central/induzido quimicamente , Doenças Autoimunes Desmielinizantes do Sistema Nervoso Central/patologia , Modelos Animais de Doenças , Camundongos , Camundongos Transgênicos , Tamoxifeno/farmacologia , Substância Branca
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