RESUMO
Female somatic X-chromosome inactivation (XCI) balances the X-linked transcriptional dosages between the sexes, randomly silencing the maternal or paternal X chromosome in each cell of 46,XX females. Skewed XCI toward one parental X has been observed in association with ageing and in some female carriers of X-linked diseases. To address the problem of non-random XCI, we quantified the XCI skew in different biological samples of naturally conceived females of different age groups and girls conceived after in vitro fertilization (IVF). Generally, XCI skew differed between saliva, blood, and buccal swabs, while saliva and blood had the most similar XCI patterns in individual females. XCI skew increased with age in saliva, but not in other tissues. We showed no significant differences in the XCI patterns in tissues of naturally conceived and IVF females. The gene expression profile of the placenta and umbilical cord blood was determined depending on the XCI pattern. The increased XCI skewing in the placental tissue was associated with the differential expression of several genes out of 40 considered herein. Notably, skewed XCI patterns (> 80:20) were identified with significantly increased expression levels of four genes: CD44, KDM6A, PHLDA2, and ZRSR2. The differences in gene expression patterns between samples with random and non-random XCI may shed new light on factors contributing to the XCI pattern outcome and indicate new paths in future research on the phenomenon of XCI skewing.
Assuntos
Placenta , Inativação do Cromossomo X , Humanos , Feminino , Gravidez , Cromossomo XRESUMO
Mucopolysaccharidoses (MPS) are a group of inherited disorders, caused by mutations in the genes coding for proteins involved (directly or indirectly) in glycosaminoglycan (GAG) degradation. A lack or drastically decreased residual activity of a GAG-degrading enzyme leads to the storage of these compounds, thus damaging proper functions of different cells, including neurons. The disease leads to serious psycho-motor dysfunctions and death before reaching the adulthood. Until now, induction of the autophagy process was considered as one of the therapeutic strategies for treatment of diseases caused by protein aggregation (Alzheimer's, Parkinson's, and Huntington's diseases). However, this strategy has only been recently suggested as a potential therapy for MPS. In this work, we show that the pharmacological stimulation of autophagy, by using valproic acid and lithium chloride, led to accelerated degradation of accumulated GAGs. Cytotoxicity tests indicated the safety of the use of the investigated compounds. We observed an increased number of lysosomes and enhanced degradation of heparan sulfate (one of GAGs). Induction of the autophagy process was confirmed by measuring abundance of the marker proteins, including LC3-II. Moreover, inhibition of this process resulted in abolition of the valproic acid- and LiCl-mediated reduction in GAG levels. This is the first report on the possibility of using valproic acid and lithium chloride for reducing levels of GAGs in neuronopathic forms of MPS.
RESUMO
Through the use of new genomic and metabolomic technologies, our comprehension of the molecular and biochemical etiologies of genetic disorders is rapidly expanding, and so are insights into their varying phenotypes. Dosage compensation (lyonization) is an epigenetic mechanism that balances the expression of genes on heteromorphic sex chromosomes. Many studies in the literature have suggested a profound influence of this phenomenon on the manifestation of X-linked disorders in females. In this review, we summarize the clinical and genetic findings in female heterozygotic carriers of a pathogenic variant in one of ten selected X-linked genes whose defects result in metabolic disorders.
Assuntos
Mecanismo Genético de Compensação de Dose/genética , Doenças Genéticas Ligadas ao Cromossomo X/genética , Doenças Metabólicas/genética , Cromossomos Humanos X/genética , Epigênese Genética/genética , Feminino , Genes Ligados ao Cromossomo X/genética , Humanos , Inativação do Cromossomo X/genéticaRESUMO
Recent studies indicated that apart from lysosomal storage of glycosaminoglycans (GAGs), secondary and tertiary changes in cellular processes may significantly contribute to development of disorders and symptoms occurring in mucopolysaccharidoses (MPS), a group of lysosomal storage diseases in which neurodegeneration is specific for most types and subtypes. In this report, using transcriptomic data, we demonstrate that regulation of hundreds of genes coding for proteins involved in regulations of various cellular processes is changed in cells derived from patients suffering from all types and subtypes of MPS. Among such genes there are 10 which expression is significantly changed in 9 or more (out of 11) MPS types/subtypes; they include IER3IP1, SAR1A, TMEM38B, PLCB4, SIN3B, ABHD5, SH3BP5, CAPG, PCOLCE2, and MN1. Moreover, there are several genes whose expression is changed over log2 > 4 times in some MPS types relative to control cells. The above analysis indicates that significant changes in expression of genes coding for various regulators of cellular processes may considerably contribute to development of cellular dysfunctions, and further appearance of specific symptoms of MPS, including neurodegeneration.
Assuntos
Microambiente Celular/fisiologia , Fibroblastos/metabolismo , Mucopolissacaridoses/genética , Mucopolissacaridoses/metabolismo , Transcriptoma/fisiologia , Linhagem Celular , Células Cultivadas , Fibroblastos/patologia , Expressão Gênica , Humanos , Mucopolissacaridoses/patologiaRESUMO
Endothelial-mesenchymal transition (EndMT) is a crucial phenomenon in regulating the development of diseases, including cancer metastasis and fibrotic disorders. The primary regulators of disease development are zinc-finger transcription factors belonging to the Snail family. In this study, we characterized the myocardin-related transcription factor (MRTF)-dependent mechanisms of a human snail promoter regulation in TGF-ß-stimulated human endothelial cells. Although in silico analysis revealed that the snail promoter's regulatory fragment contains one GCCG and two SP1 motifs that could be occupied by MRTFs, the genetic study confirmed that MRTF binds only to SP1 sites to promote snail expression. The more accurate studies revealed that MRTF-A binds to both SP1 elements, whereas MRTF-B to only one (SP1near). Although we found that each MRTF alone is capable of inducing snail expression, the direct cooperation of these proteins is required to reinforce snail expression and promote the late stages of EndMT within 48 hours. Furthermore, genetic and biochemical analysis revealed that MRTF-B alone could induce the late stage of EndMT. However, it requires a prolonged time. Therefore, we concluded that MRTFs might cause EndMT in a fast- and slow-dependent manner. Based on MRTF-dependent Snail upregulation, we recognized that TGF-ß1, as an MRTF-B regulator, is involved in slow EndMT induction, whereas TGF-ß2, which altered both MRTF-A and MRTF-B expression, promotes a fast EndMT process.
Assuntos
Transição Epitelial-Mesenquimal , Fatores de Transcrição da Família Snail/genética , Fatores de Transcrição/metabolismo , Linhagem Celular , Células Endoteliais/metabolismo , Células Endoteliais/patologia , Humanos , Regiões Promotoras Genéticas , Ligação Proteica , Fatores de Transcrição da Família Snail/metabolismo , Ativação TranscricionalRESUMO
BACKGROUND: The lifestyle of young boys has impact on the risks of cardiovascular diseases. OBJECTIVE: The aim of the study was to evaluate the effect of atherosclerosis risk factors determined by overweight and obesity and lifestyle, i.e.: eating habits, low physical activity and smoking cigarettes, on blood lipid profile of boys at the age of 16 to 18. MATERIAL AND METHODS: The study covered 369 boys from secondary schools. They were evaluated for the supply of dietary constituents with atherogenic and protective actions, for nutritional status acc. to Cole's criteria, the level of physical activity, and smoking cigarettes. Lipid metabolism was determined based on criteria recommended by the American National Cholesterol Education Program (NCEP). Logistic regression analysis was conducted and risk odds ratio [OR] was determined. RESULTS: Analyses showed the boys to be characterized by overweight (10.8%) and obesity (2.7%), and by inappropriate concentration of total cholesterol (26.5%), LDL (13.3%), HDL (21.7%) and triglycerides (41.7%). High BMI turned out to be a significant risk factor of an elevated total cholesterol concentration: [aOR]=2.27; triglycerides: [aOR]=2.35 and LDL: [aOR]=2.41. Low physical activity was found to negatively affect the concentration of LDL: [aOR]=1.88. The boys smoking cigarettes were shown to have a reduced HDL: [aOR]=1.65. The total content of fat and saturated fatty acids in diet exerted a significantly negative impact on blood lipid profile of the boys. CONCLUSIONS: The lifestyle of the young boys was demonstrated to determine the risk of cardiovascular diseases. Overweight and obesity, abdominal obesity in particular, were found to be a significant risk factor of disorders in their lipid metabolism.
Assuntos
Exercício Físico , Comportamento Alimentar , Produtos do Tabaco , Adolescente , Índice de Massa Corporal , Humanos , Lipídeos , Masculino , Polônia , Fatores de Risco , Fumar/epidemiologiaRESUMO
Background: A recently growing number of children and adolescents with overweight or obesity is indicative of the need for diagnosing their complications that may appear in the early childhood. For this reason, diagnostic criteria were developed for components of the metabolic syndrome (MS) also for these groups of the population. Objective: This study was aimed at evaluating the frequency of metabolic syndrome risk factors occurrence in children and adolescents from the city of Wroclaw and surroundings depending on gender, age and physical activity. Material and methods: Investigations on the frequency of occurrence of dietary and non-dietary risk factors of the metabolic syndrome were carried out in the years 2010-2017 among 771 children and adolescents aged 10-18 year, attending to primary schools, gymnasiums and secondary schools in Wroclaw. Results: The lack of any components of the metabolic syndrome was demonstrated in 14.78% of the children aged 10-12 years as well as in 17.38% of both adolescents aged 13-15 and 17-18 years. One risk factor was most frequently diagnosed in children aged 10-12 years (17.89%) and it was arterial hypertension (16.08% of the whole surveyed population). Three MS components were demonstrated in 15 persons (1.95% of the whole surveyed population), including in 7 girls and 8 boys. The persons with three MS risk factors from the age category 13-15 years constituted 0.26% whereas these from the age category 17-18 years constituted 1.69% of the whole surveyed group. Conclusions: It shall be concluded that the incidence of the fully symptomatic MS depended significantly on the age of the surveyed, but not on their gender. Among the three adopted components of MS, the most frequently demonstrated disorders included: arterial hypertension, abdominal obesity and increased concentration of triglycerides in blood serum.
Assuntos
Exercício Físico/fisiologia , Síndrome Metabólica/diagnóstico , Síndrome Metabólica/fisiopatologia , Obesidade Infantil/diagnóstico , Obesidade Infantil/fisiopatologia , Adolescente , Fatores Etários , Criança , Feminino , Humanos , Masculino , Síndrome Metabólica/epidemiologia , Obesidade Infantil/epidemiologia , Polônia/epidemiologia , Prevalência , Fatores de Risco , Fatores SexuaisRESUMO
Obstacles to effective therapies for mucopolysaccharidoses (MPSs) determine the need for continuous studies in order to enhance therapeutic strategies. Dimethyl sulfoxide (DMSO) is frequently utilised as a solvent in biological studies, and as a vehicle for drug therapy and the in vivo administration of water-insoluble substances. In the light of the uncertainty on the mechanisms of DMSO impact on metabolism of glycosaminoglycans (GAGs) pathologically accumulated in MPSs, in this work, we made an attempt to investigate and resolve the question of the nature of GAG level modulation by DMSO, the isoflavone genistein solvent employed previously by our group in MPS treatment. In this work, we first found the cytotoxic effect of DMSO on human fibroblasts at concentrations above 3%. Also, our results displayed the potential role of DMSO in the regulation of biological processes at the transcriptional level, then demonstrated a moderate impact of the solvent on GAG synthesis. Interestingly, alterations of lysosomal ultrastructure upon DMSO treatment were visible. As there is growing evidence in the literature that DMSO can affect cellular pathways leading to numerous changes, it is important to expand our knowledge concerning this issue.
Assuntos
Dimetil Sulfóxido/administração & dosagem , Genisteína/administração & dosagem , Doenças por Armazenamento dos Lisossomos/tratamento farmacológico , Mucopolissacaridoses/tratamento farmacológico , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Glicosaminoglicanos/antagonistas & inibidores , Humanos , Isoflavonas/metabolismo , Doenças por Armazenamento dos Lisossomos/metabolismo , Doenças por Armazenamento dos Lisossomos/patologia , Lisossomos/efeitos dos fármacos , Lisossomos/ultraestrutura , Mucopolissacaridoses/metabolismo , Mucopolissacaridoses/patologiaRESUMO
Epigenetic mechanisms, including histone post-translational modifications, are central regulators of cell cycle control. The euchromatic G9a histone methyltransferase (G9a HMT) is a key enzyme catalyzing histone H3 methylation on lysines 9 and 27, and its dysregulation has been linked to uncontrolled proliferation of tumor cells. Here, we have investigated the effect of G9a HMT silencing on cell proliferation of microvascular endothelial cells, a process necessary to sustain tumor growth through the formation of the vascular capillary network. Inhibition of G9a HMT activity in human microvascular endothelial cells (HMEC-1) was performed either pharmacologically, by treatment of cells with BIX-01294 or chaetocin, or transcriptionally, using shRNA. Cell viability and proliferation were examined using the resazurin reduction assay, flow cytometry and immunostaining of phosphorylated checkpoint kinase 1 (pSer317Chk1). Expression of cell cycle- and redox homeostasis-related genes was determined by quantitative PCR. Reactive oxygen species production was measured by oxidation of the fluorescent probe 2',7'-dichlorodihydrofluorescein diacetate and the cell's total antioxidant capacity by using the ABTS assay. Inhibition of G9a HMT activity by BIX-01294 treatment or by shRNA attenuated the proliferation of HMEC-1, nuclear localization of phosphorylated Chk1, and induced cell cycle arrest in G1 phase. Transcriptional analysis demonstrated increased gene expression of the cyclin-dependent kinase (CDK) inhibitor p21, and also of Rb1, in BIX-01294 treated cells. Decreased proliferation rate was accompanied by enhanced antioxidant potential of HMEC-1 cells, as demonstrated by reduced production of reactive oxygen species, increased total antioxidant capacity and expression of the antioxidant enzymes catalase and superoxide dismutase 1. Collectively, our results demonstrate of the central role of G9a HMT in the promotion of endothelial cells proliferation, and suggest that endothelial G9a HMT may be a target in the treatment of vascular proliferative disorders and tumor neovascularization.
Assuntos
Proliferação de Células/fisiologia , Células Endoteliais/fisiologia , Antígenos de Histocompatibilidade/fisiologia , Histona-Lisina N-Metiltransferase/fisiologia , Microvasos/citologia , Azepinas/farmacologia , Linhagem Celular , Proliferação de Células/efeitos dos fármacos , Células Endoteliais/efeitos dos fármacos , Antígenos de Histocompatibilidade/genética , Histona-Lisina N-Metiltransferase/antagonistas & inibidores , Histona-Lisina N-Metiltransferase/genética , Homeostase , Humanos , Oxirredução , Quinazolinas/farmacologia , RNA Interferente Pequeno/genéticaRESUMO
Sanfilippo disease is one of mucopolysaccharidoses (MPS), a group of lysosomal storage diseases characterized by accumulation of partially degraded glycosaminoglycans (GAGs). It is classified as MPS type III, though it is caused by four different genetic defects, determining subtypes A, B, C and D. In each subtype of MPS III, the primary storage GAG is heparan sulfate (HS), but mutations leading to A, B, C, and D subtypes are located in genes coding for heparan N-sulfatase (the SGSH gene), α-N-acetylglucosaminidase (the NAGLU gene), acetyl-CoA:α-glucosaminide acetyltransferase (the HGSNAT gene), and N-acetylglucosamine-6-sulfatase (the GNS gene), respectively. Neurodegenerative changes in the central nervous system (CNS) are major problems in Sanfilippo disease. They cause severe cognitive disabilities and behavioral disturbances. This is the main reason of a current lack of therapeutic options for MPS III patients, while patients from some other MPS types (I, II, IVA, and VI) can be treated with enzyme replacement therapy or bone marrow or hematopoietic stem cell transplantations. Nevertheless, although no therapy is available for Sanfilippo disease now, recent years did bring important breakthroughs in this aspect, and clinical trials are being conducted with enzyme replacement therapy, gene therapy, and substrate reduction therapy. These recent achievements are summarized and discussed in this review.
Assuntos
Terapia Genética , Transplante de Células-Tronco Hematopoéticas , Heparitina Sulfato/uso terapêutico , Sulfatases/uso terapêutico , Acetilglucosaminidase/genética , Terapia Genética/métodos , Humanos , Mutação/genéticaRESUMO
Autophagy is a process of degradation of macromolecules in the cytoplasm, particularly proteins of a long half-life, as well as whole organelles, in eukaryotic cells. Lysosomes play crucial roles during this degradation. Autophagy is a phylogenetically old, and evolutionarily conserved phenomenon which occurs in all eukaryotic cells. It can be found in yeast Saccharomyces cerevisiae, insect Drosophila melanogaster, and mammals, including humans. Its high importance for cell physiology has been recognized, and in fact, dysfunctions causing impaired autophagy are associated with many severe disorders, including cancer and metabolic brain diseases. The types and molecular mechanisms of autophagy have been reviewed recently by others, and in this paper they will be summarized only briefly. Regulatory networks controlling the autophagy process are usually described as negative regulations. In contrast, here, we focus on different ways by which autophagy can be stimulated. In fact, activation of this process by different factors or processes can be considered as a therapeutic strategy in metabolic neurodegenerative diseases. These aspects are reviewed and discussed in this article.
Assuntos
Autofagia/efeitos dos fármacos , Doenças Neurodegenerativas/tratamento farmacológico , Fármacos Neuroprotetores/uso terapêutico , Animais , Humanos , Lisossomos/metabolismo , Doenças Neurodegenerativas/metabolismo , Fármacos Neuroprotetores/farmacologia , Transdução de Sinais/efeitos dos fármacosRESUMO
Oxidative stress may be the major cause of induction of Shiga toxin-converting (Stx) prophages from chromosomes of Shiga toxin-producing Escherichia coli (STEC) in human intestine. Thus, we aimed to test a series of novel antioxidant compounds for their activities against prophage induction, thus, preventing pathogenicity of STEC. Forty-six compounds (derivatives of carbazole, indazole, triazole, quinolone, ninhydrine, and indenoindole) were tested. Fifteen of them gave promising results and were further characterized. Eleven compounds had acceptable profiles in cytotoxicity tests with human HEK-293 and HDFa cell lines. Three of them (selected for molecular studies) prevent the prophage induction at the level of expression of specific phage genes. In bacterial cells treated with hydrogen peroxide, expression of genes involved in the oxidative stress response was significantly less efficient in the presence of the tested compounds. Therefore, they apparently reduce the oxidative stress, which prevents induction of Stx prophage in E. coli.
Assuntos
Antibacterianos/farmacologia , Antioxidantes/farmacologia , Toxina Shiga/antagonistas & inibidores , Escherichia coli Shiga Toxigênica/efeitos dos fármacos , Antibacterianos/síntese química , Antibacterianos/química , Antioxidantes/síntese química , Antioxidantes/química , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Células HEK293 , Humanos , Estrutura Molecular , Estresse Oxidativo/efeitos dos fármacos , Toxina Shiga/genética , Toxina Shiga/metabolismo , Escherichia coli Shiga Toxigênica/citologia , Escherichia coli Shiga Toxigênica/metabolismo , Relação Estrutura-AtividadeRESUMO
Genetic and immunological diseases, despite many attempts to develop effective treatments, still remain a great challenge for medicine. Current therapies of these diseases consist of pharmacological alleviation of symptoms, rehabilitation and psychological help which, although very important, are not sufficient. Therefore, searching for new therapeutics which could remove the major causes of these diseases is of particular importance for the society. Natural compounds reveal many biological activities which makes them candidates for drugs in such diseases. One of them is genistein, a compound from the group of flavonoids. As it affects multiple processes, genistein has become in the center of interest of many scientists working on diseases of various etiology, course and inheritance. It was used in experimental therapies of some genetic diseases (Huntington's disease, amyotrophic lateral sclerosis Parkinson disease, cystic fibrosis), as well as autoimmunological diseases and allergies. Clinical trials with the use of genistein in treatment of patients suffering from Alzheimer's diseases and mucopolysaccharidosis type III are ongoing. The employment of differential properties of genistein in attempts to treat each of these diseases is of special interest. In this review, detailed molecular mechanisms of genistein action are summarized in the light of therapies of the above mentioned genetic and immunological diseases, including description of therapeutic potentials of each activity of this isoflavone, efficiency of its action, and its potential use as a drug in the future.
Assuntos
Genisteína/farmacologia , Genisteína/uso terapêutico , Doenças do Sistema Imunitário/tratamento farmacológico , Doença de Alzheimer/tratamento farmacológico , Doença de Alzheimer/genética , Esclerose Lateral Amiotrófica/tratamento farmacológico , Esclerose Lateral Amiotrófica/genética , Humanos , Doença de Huntington/tratamento farmacológico , Doença de Huntington/genética , Mucopolissacaridose III/tratamento farmacológico , Doença de Parkinson/tratamento farmacológico , Doença de Parkinson/genéticaRESUMO
The aim of the present study was to investigate serum levels of novel markers: interleukin 17A (IL-17A), anaphylatoxin C5a and chemokine regulated upon activation normal T-cell expressed and secreted (RANTES) in neonates with clinically suspected early-onset neonatal sepsis (EONS), and to compare their values with those of non-infected neonates. Eighteen neonates with clinical signs and symptoms of EONS were enrolled in this study. Fifty healthy, non-infected neonates served as the control group. In all neonates serum levels of IL-17A, C5a and RANTES were measured by solid-phase sandwich enzyme-linked immunosorbent assay (ELISA). At the time of investigation serum levels of anaphylatoxin C5a were significantly higher in neonates with clinical symptoms of EONS than in non-infected neonates (median 65.35 vs. 50.4 ng/ml, p = 0.034), whereas levels of RANTES were similar and levels of IL-17A were under detection limit of the method. Based on these preliminary results, serum levels of C5a may be a useful marker of inflammation in early onset neonatal sepsis. Because traditional methods of microbiological diagnostics in EONS are frequently unsuccessful, the search for an alternative laboratory biomarkers is of great clinical importance. Thus, there is a strong need for further studies evaluating usefulness of this anaphylatoxin in EONS diagnosis on a larger group of patients.
RESUMO
Flavonoids have been studied as potential agents in medicine for many years. Among them, genistein was found to be active in various biological systems, mainly in prevention of cancer. Our recent work supported the idea that genistein also impacts multiple cellular processes in healthy fibroblasts; however, its effects on cell cycle-related pathways remained to be elucidated. Thus, in this work, high throughput screening with microarrays coupled to real-time quantitative Reverse Transcription PCR analyses was employed to study the changes in expression of key genes associated with cell cycle regulation and/or DNA replication in response to genistein, kaempferol, daidzein, and mixtures of genistein and either kaempferol or daidzein. Among them, genistein was found as the most significantly modulating, in a time- and dose-dependent manner, compound of activity of studied genes, whose products are involved in different phases of the cell cycle and/or in regulatory processes important for DNA replication and cell growth. It considerably reduced the efficiency of expression of genes coding for MCM2-7 and MCM10 helicases, as well as some other proteins involved in the S phase control. In addition, genistein caused cell cycle arrest in the G2/M phase, which was accompanied by activation of CDKN1A, CDKN1C, CDKN2A, CDKN2B, CDKN2C, and GADD45A genes, as well as down-regulation of several mRNAs specific for this stage, demonstrated by transcriptomic assessments. We believe that studies described in this paper will be helpful in elucidating molecular mechanisms of action of genistein as modulator of cell cycle and inhibitor of DNA replication in humans.
Assuntos
Ciclo Celular/efeitos dos fármacos , Replicação do DNA/efeitos dos fármacos , Fibroblastos/efeitos dos fármacos , Flavonoides/farmacologia , Células Cultivadas , Fibroblastos/citologia , Perfilação da Expressão Gênica , Regulação da Expressão Gênica/efeitos dos fármacos , Genisteína/farmacologia , Humanos , Isoflavonas/farmacologia , Quempferóis/farmacologia , Proteínas de Manutenção de Minicromossomo/genética , Análise de Sequência com Séries de OligonucleotídeosRESUMO
BACKGROUND: Youth nutrition and their nutritional status are conditioned by many factors, some of the main ones being: economic, social, climatic, cultural, and psychological factors as well as nutritional knowledge. With the growing problem of overweight and obesity among children and young people, the incidence of the metabolic syndrome is also increasing. OBJECTIVES: The aim of the study was to assess the impact of demographic, sociological and psychological factors on the incidence of obesity among 17-18-year-old adolescents from Wroclaw and vicinity as a major risk factor for the development of the metabolic syndrome. MATERIAL AND METHODS: The study was conducted in three upper-secondary schools in Wroclaw, Poland. In the surveyed group (17-18 years old, n = 269) girls accounted for 59.5% and boys constituted 40.5%. Majority of young people were Wroclaw citizens (72.9%). Centile charts elaborated by the Children's Memorial Health Institute were adopted for the evaluation of anthropometric parameters. Evaluation of the impact of non-dietary factors on the manner of nutrition was carried out using own questionnaire. RESULTS: Based on the tests, abdominal obesity was determined among 34.5% of adolescents aged 17 years and among 65.5% of these aged 18 years. Obesity was more common in girls carrying genetic burden of the disease. Youth with the largest waist circumference most often declared to use slimming diets - 6.7%, and the lowest hunger sensation in stress - 3.4%. In addition, 30.5% of the adolescents with the smallest waist circumference and 11.5% with the largest waist circumference declared to be non-smoking. Occasional alcohol consumption was declared by 30.1% of young people with the smallest waist circumference, and 13.4% with the largest waist circumference. CONCLUSIONS: Youth with abdominal obesity significantly more likely than those with normal waist circumference applied slimming diets. Significant impact on the formation of abdominal obesity among girls had inherited disease burden.
Assuntos
Dieta/estatística & dados numéricos , Comportamentos Relacionados com a Saúde , Síndrome Metabólica/epidemiologia , Obesidade Abdominal/epidemiologia , Obesidade Infantil/epidemiologia , Adolescente , Índice de Massa Corporal , Dieta/psicologia , Comportamento Alimentar , Feminino , Humanos , Masculino , Síndrome Metabólica/psicologia , Obesidade Abdominal/psicologia , Obesidade Infantil/psicologia , Grupo Associado , Polônia/epidemiologia , Prevalência , Distribuição por Sexo , Fatores Socioeconômicos , Circunferência da CinturaRESUMO
A growing number of studies confirm an important effect of diet, lifestyle and physical activity on health status, the ageing process and many metabolic disorders. This study focuses on the influence of a diet supplement, NucleVital®Q10 Complex, on parameters related to redox homeostasis and ageing. An experimental group of 66 healthy volunteer women aged 35-55 supplemented their diet for 12 weeks with the complex, which contained omega-3 acids (1350 mg/day), ubiquinone (300 mg/day), astaxanthin (15 mg/day), lycopene (45 mg/day), lutein palmitate (30 mg/day), zeaxanthine palmitate (6 mg/day), L-selenomethionine (330 mg/day), cholecalciferol (30 µg/day) and α-tocopherol (45 mg/day). We found that NucleVital®Q10 Complex supplementation significantly increased total antioxidant capacity of plasma and activity of erythrocyte superoxide dismutase, with slight effects on oxidative stress biomarkers in erythrocytes; MDA and 4-hydroxyalkene levels. Apart from the observed antioxidative effects, the tested supplement also showed anti-ageing activity. Analysis of expression of SIRT1 and 2 in PBMCs showed significant changes for both genes on a mRNA level. The level of telomerase was also increased by more than 25%, although the length of lymphocyte telomeres, determined by RT-PCR, remained unchanged. Our results demonstrate beneficial effects concerning the antioxidant potential of plasma as well as biomarkers related to ageing even after short term supplementation of diet with NucleVital®Q10 Complex.
Assuntos
Envelhecimento/efeitos dos fármacos , Antioxidantes/administração & dosagem , Micronutrientes/administração & dosagem , Adulto , Biomarcadores/sangue , Fator Neurotrófico Derivado do Encéfalo/sangue , Suplementos Nutricionais , Combinação de Medicamentos , Eritrócitos/enzimologia , Feminino , Glutationa Peroxidase/sangue , Homeostase , Humanos , Leucócitos Mononucleares/fisiologia , Peroxidação de Lipídeos , Pessoa de Meia-Idade , Oxirredução , Estresse Oxidativo , Superóxido Dismutase/sangue , Homeostase do Telômero/efeitos dos fármacosRESUMO
BACKGROUND: Hyaluronan (HA) is an extracellular glycosaminoglycan polysaccharide with widespread roles throughout development and in healthy and neoplastic tissues. In pluripotent stem cell culture it can support both stem cell renewal and differentiation. However, responses to HA in culture are influenced by interaction with a range of cognate factors and receptors including components of blood serum supplements, which alter results. These may contribute to variation in cell batch production yield and phenotype as well as heighten the risks of adventitious pathogen transmission in the course of cell processing for therapeutic applications. MAIN: Here we characterise differentiation of a human embryo/pluripotent stem cell derived Mesenchymal Stromal Cell (hESC/PSC-MSC)-like cell population by culture on a planar surface coated with HA in serum-free media qualified for cell production for therapy. Resulting cells met minimum criteria of the International Society for Cellular Therapy for identification as MSC by expression of. CD90, CD73, CD105, and lack of expression for CD34, CD45, CD14 and HLA-II. They were positive for other MSC associated markers (i.e.CD166, CD56, CD44, HLA 1-A) whilst negative for others (e.g. CD271, CD71, CD146). In vitro co-culture assessment of MSC associated functionality confirmed support of growth of hematopoietic progenitors and inhibition of mitogen activated proliferation of lymphocytes from umbilical cord and adult peripheral blood mononuclear cells, respectively. Co-culture with immortalized THP-1 monocyte derived macrophages (Mɸ) concurrently stimulated with lipopolysaccharide as a pro-inflammatory stimulus, resulted in a dose dependent increase in pro-inflammatory IL6 but negligible effect on TNFα. To further investigate these functionalities, a bulk cell RNA sequence comparison with adult human bone marrow derived MSC and hESC substantiated a distinctive genetic signature more proximate to the former. CONCLUSION: Cultivation of human pluripotent stem cells on a planar substrate of HA in serum-free culture media systems is sufficient to yield a distinctive developmental mesenchymal stromal cell lineage with potential to modify the function of haematopoietic lineages in therapeutic applications.
Assuntos
Diferenciação Celular , Ácido Hialurônico , Células-Tronco Mesenquimais , Células-Tronco Pluripotentes , Humanos , Células-Tronco Mesenquimais/metabolismo , Células-Tronco Mesenquimais/citologia , Ácido Hialurônico/farmacologia , Ácido Hialurônico/metabolismo , Células-Tronco Pluripotentes/metabolismo , Células-Tronco Pluripotentes/citologia , Meios de Cultura Livres de Soro/farmacologia , Linhagem da Célula , Células Cultivadas , Técnicas de Cultura de Células/métodos , Técnicas de CoculturaRESUMO
Mucopolysaccharidosis type VI (MPS VI) is a rare lysosomal, autosomal recessive storage disorder caused by deficient activity of N-acetylgalactosamine-4-sulfatase (ARSB). Approximately, 140 ARSB gene mutations have been identified; however, most are private mutations making genotype-phenotype correlation for most MPS VI patients difficult. The aim of this study was to describe the natural clinical course in patients homozygous for the p.R152W mutation from eight unrelated families. From our database of 70 patients with MPS VI, we selected 10 patients homozygous for the p.R152W mutant allele (median age 27.5 years, range 18-38 years). We performed a cross-sectional observational study characterizing the onset and prevalence of clinical manifestations. First signs of the disease, such as cardiac valve disease, slightly decreased joint range of motion and mild growth retardation, were observed in mid-adolescent years (median 15 years). Within the disease course, the most common clinical feature in all the patients was progressive heart disease of predominantly valve origin leading to symptoms of heart failure. Other typical MPS VI features were subtle and not present in all the patients. Delays up to 23 years (median 8.5 years) intervened between symptom onset and disease diagnosis. Patients homozygous for the p.R152W mutation present a cardiac variant of MPS VI characterized by progressive cardiac valve disease leading to serious cardiac complications including abrupt death due to cardiac failure.
Assuntos
Insuficiência Cardíaca/genética , Mucopolissacaridose VI/genética , N-Acetilgalactosamina-4-Sulfatase/genética , Polimorfismo Genético/genética , Adolescente , Adulto , Substituição de Aminoácidos , Estudos de Coortes , Estudos Transversais , Demografia , Europa (Continente) , Feminino , Frequência do Gene , Estudos de Associação Genética , Insuficiência Cardíaca/diagnóstico , Valvas Cardíacas/fisiopatologia , Homozigoto , Humanos , Masculino , Moscou , Mucopolissacaridose VI/diagnóstico , Mutação , Fenótipo , Adulto JovemRESUMO
Mucopolysaccharidosis VI (MPS VI) is a rare autosomal recessive disorder caused by a deficiency of N-acetylgalactosamine-4-sulfatase (ARSB). Over 130 ARSB gene mutations have been identified thus far and most mutations are unique to individual families. We aimed to analyze the spectrum of mutations in the ARSB gene responsible for the disorder in Poland, Belarus and Baltic States. Twenty one families with MPS VI patients, in whom diagnosis was confirmed biochemically and enzymatically, were studied. Direct sequencing of patient genomic DNA was used to identify ARSB mutations. In total, fourteen different disease-causing mutations were found. Three novel mutations included insertion c.375_376insT, a missense mutation c.499G>A (p.G167R) and deletion/insertion c.750_754delinsCCTGAAGTCAAG. We also report 11 previously described mutations (p.A33V, p.W57C, p.Q88X, p.T92K, p.Q97X, p.R152W, p.R160Q, p.R160X, p.Y210C, p.Y266S, p.G302R). The mutation p.R152W was present at a high prevalence of 50% (21/42) the mutated alleles in this group of patients. High prevalence of p.R152W mutation in Poland, Belarus and Baltic States indicates a possible founder effect and suggests that screening for this mutation may be appropriate in MPS VI patients from this region. Our study has also provided evidence to support genotype-phenotype correlation.