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1.
Int J Mol Sci ; 22(15)2021 Jul 28.
Artigo em Inglês | MEDLINE | ID: mdl-34360820

RESUMO

We designed a concept of 3D-printed attachment with porous glass filter disks-SLIDE (Sweat sampLIng DevicE) for easy sampling of apocrine sweat. By applying advanced mass spectrometry coupled with the liquid chromatography technique, the complex lipid profiles were measured to evaluate the reproducibility and robustness of this novel approach. Moreover, our in-depth statistical evaluation of the data provided an insight into the potential use of apocrine sweat as a novel and diagnostically relevant biofluid for clinical analyses. Data transformation using probabilistic quotient normalization (PQN) significantly improved the analytical characteristics and overcame the 'sample dilution issue' of the sampling. The lipidomic content of apocrine sweat from healthy subjects was described in terms of identification and quantitation. A total of 240 lipids across 15 classes were identified. The lipid concentrations varied from 10-10 to 10-4 mol/L. The most numerous class of lipids were ceramides (n = 61), while the free fatty acids were the most abundant ones (average concentrations of 10-5 mol/L). The main advantages of apocrine sweat microsampling include: (a) the non-invasiveness of the procedure and (b) the unique feature of apocrine sweat, reflecting metabolome and lipidome of the intracellular space and plasmatic membranes. The SLIDE application as a sampling technique of apocrine sweat brings a promising alternative, including various possibilities in modern clinical practice.


Assuntos
Lipidômica/métodos , Lipídeos/análise , Metabolômica/métodos , Manejo de Espécimes , Suor/química , Voluntários Saudáveis , Humanos
2.
Molecules ; 26(13)2021 Jul 03.
Artigo em Inglês | MEDLINE | ID: mdl-34279415

RESUMO

Neurodegeneration is the consequence of harmful events affecting the nervous system that lead to neuronal death. Toxic substances, including air pollutants, are capable of inducing neurodegeneration. Ozone (O3) is the most oxidative toxic pollutant. O3 reacts with cellular components and forms reactive oxygen and nitrogen species, triggering nitro-oxidative damage during short-term exposure. Curcumin (CUR) is a natural phenolic molecule bearing well-documented antioxidant and anti-inflammatory biological activities in diverse experimental models. The aim of this work was to evaluate the effect of preventive dietary administration of CUR against hippocampal neurodegeneration and nitro-oxidative damage caused by short-term exposure to O3. Eighty Wistar male rats were distributed into four experimental groups, twenty rats each: intact control; CUR dietary supplementation without O3 exposure; exposure to 0.7 ppm of O3; and exposed to O3 with CUR dietary supplementation. Five rats from each group were sacrificed at 1, 2, 4, and 8 h of exposure. The CUR dose was 5.6 mg/kg and adjusted according to food consumption. CUR significantly decreased oxidative damage to plasma lipids and proteins, as well as neurodegeneration in CA1 and CA3 hippocampal regions. Concluding, CUR proved effective protection in decreasing neurodegeneration in the hippocampus and prevented systemic oxidative damage.


Assuntos
Proteínas Sanguíneas/metabolismo , Curcumina/farmacologia , Hipocampo/efeitos dos fármacos , Lipídeos/análise , Doenças Neurodegenerativas/tratamento farmacológico , Estresse Oxidativo , Ozônio/toxicidade , Animais , Anti-Inflamatórios não Esteroides/farmacologia , Antioxidantes/farmacologia , Hipocampo/metabolismo , Hipocampo/patologia , Masculino , Doenças Neurodegenerativas/induzido quimicamente , Doenças Neurodegenerativas/metabolismo , Doenças Neurodegenerativas/patologia , Óxido Nítrico/metabolismo , Ratos , Ratos Wistar
3.
Molecules ; 26(14)2021 Jul 07.
Artigo em Inglês | MEDLINE | ID: mdl-34299399

RESUMO

Celastrol, a pentacyclic triterpene isolated from the traditional Chinese medicine Tripterygium wilfordii Hook. F., exhibits effectiveness in protection against multiple central nervous system (CNS) diseases such as cerebral ischemia, but its influence on lipidomics still remains unclear. Therefore, in the present study, the efficacy and potential mechanism of celastrol against cerebral ischemia/reperfusion (I/R) injury were investigated based on lipidomics. Middle cerebral artery occlusion (MCAO) followed by reperfusion was operated in mice to set up a cerebral I/R model. TTC staining and TUNEL staining were used to evaluate the therapeutic effect of celastrol. Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC/MS) was employed for lipidomics analysis in ipsilateral hemisphere and plasma. Celastrol remarkably reduced cerebral infarct volume and apoptosis positive cells in tMCAO mice. Furthermore, lipidomics analysis showed that 14 common differentially expressed lipids (DELs) were identified in brain and five common DELs were identified in plasma between the Sham, tMCAO and Celastrol-treated tMCAO groups. Through enrichment analysis, sphingolipid metabolism and glycerophospholipid metabolism were demonstrated to be significantly enriched in all the comparison groups. Among the DELs, celastrol could reverse cerebral I/R injury-induced alteration of phosphatidylcholine, phosphatidylethanolamine and sulfatide, which may be responsible for the neuroprotective effect of celastrol. Our findings suggested the neuroprotection of celastrol on cerebral I/R injury may be partially associated with its regulation of lipid metabolism.


Assuntos
Isquemia Encefálica/metabolismo , Encéfalo/metabolismo , Modelos Animais de Doenças , Infarto da Artéria Cerebral Média/complicações , Lipídeos/análise , Triterpenos Pentacíclicos/farmacologia , Traumatismo por Reperfusão/metabolismo , Animais , Encéfalo/efeitos dos fármacos , Isquemia Encefálica/tratamento farmacológico , Isquemia Encefálica/etiologia , Isquemia Encefálica/patologia , Lipidômica/métodos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Fármacos Neuroprotetores/farmacologia , Traumatismo por Reperfusão/tratamento farmacológico , Traumatismo por Reperfusão/etiologia , Traumatismo por Reperfusão/patologia
4.
Molecules ; 26(14)2021 Jul 07.
Artigo em Inglês | MEDLINE | ID: mdl-34299415

RESUMO

Chemical characteristics of raw and processed Istarski prsut (PDO) produced from two different pig genotypes were studied with special emphasis on amino and fatty acid composition and factors of lipid stability. Raw hams of Large White (LW)xLandrace (L), and (LWxL)xDuroc (D) pig genotypes were used in the study (20 hams of each genotype). All left raw hams from each carcass were processed in accordance with the PDO specification of Istarski prsut, and other half (the right ones) of LWxL)xD genotype were used for analyses of raw hams (fresh muscles). Istarski prsut was evaluated on the basis of the chemical parameters of the raw and matured lean ham. The process of dry curing significantly influenced the chemical properties of Istarski prsut. Despite the higher content of intramuscular fat and polyunsaturated fatty acids, the fat of (LWxL)xD ham was much more resistant to hydrolysis and oxidation, suggesting that fatty acid profile and other factors, also play a significant role. Significant differences between pig genotypes in the amino acid and fatty acid profiles were found. The analyzed Istarski prsut may be distinguished by prints of multivariate chemometric statistical analysis, based on their amino acid and fatty acid compositions.


Assuntos
Manipulação de Alimentos/métodos , Lipídeos/análise , Produtos da Carne/análise , Músculo Esquelético/crescimento & desenvolvimento , Carne de Porco/análise , Suínos/crescimento & desenvolvimento , Animais , Fenômenos Químicos , Genótipo , Músculo Esquelético/metabolismo , Suínos/classificação , Suínos/genética
5.
Int J Mol Sci ; 22(14)2021 Jul 12.
Artigo em Inglês | MEDLINE | ID: mdl-34299070

RESUMO

The aim of the current study was to test the hypothesis that maternal lipid metabolism was modulated during normal pregnancy and that these modulations are altered in gestational diabetes mellitus (GDM). We tested this hypothesis using an established mouse model of diet-induced obesity with pregnancy-associated loss of glucose tolerance and a novel lipid analysis tool, Lipid Traffic Analysis, that uses the temporal distribution of lipids to identify differences in the control of lipid metabolism through a time course. Our results suggest that the start of pregnancy is associated with several changes in lipid metabolism, including fewer variables associated with de novo lipogenesis and fewer PUFA-containing lipids in the circulation. Several of the changes in lipid metabolism in healthy pregnancies were less apparent or occurred later in dams who developed GDM. Some changes in maternal lipid metabolism in the obese-GDM group were so late as to only occur as the control dams' systems began to switch back towards the non-pregnant state. These results demonstrate that lipid metabolism is modulated in healthy pregnancy and the timing of these changes is altered in GDM pregnancies. These findings raise important questions about how lipid metabolism contributes to changes in metabolism during healthy pregnancies. Furthermore, as alterations in the lipidome are present before the loss of glucose tolerance, they could contribute to the development of GDM mechanistically.


Assuntos
Diabetes Gestacional/patologia , Metabolismo dos Lipídeos , Lipidômica/métodos , Lipídeos/análise , Obesidade/fisiopatologia , Animais , Glicemia/análise , Diabetes Gestacional/etiologia , Diabetes Gestacional/metabolismo , Feminino , Teste de Tolerância a Glucose , Camundongos , Gravidez
6.
Int J Mol Sci ; 22(14)2021 Jul 12.
Artigo em Inglês | MEDLINE | ID: mdl-34299079

RESUMO

In order to tackle the study of DNA repair pathways, the physical and chemical agents creating DNA damage, the genotoxins, are frequently employed. Despite their utility, their effects are rarely restricted to DNA, and therefore simultaneously harm other cell biomolecules. Methyl methanesulfonate (MMS) is an alkylating agent that acts on DNA by preferentially methylating guanine and adenine bases. It is broadly used both in basic genome stability research and as a model for mechanistic studies to understand how alkylating agents work, such as those used in chemotherapy. Nevertheless, MMS exerts additional actions, such as oxidation and acetylation of proteins. In this work, we introduce the important notion that MMS also triggers a lipid stress that stems from and affects the inner nuclear membrane. The inner nuclear membrane plays an essential role in virtually all genome stability maintenance pathways. Thus, we want to raise awareness that the relative contribution of lipid and genotoxic stresses when using MMS may be difficult to dissect and will matter in the conclusions drawn from those studies.


Assuntos
Antineoplásicos Alquilantes/efeitos adversos , Dano ao DNA , Lipídeos/análise , Metanossulfonato de Metila/efeitos adversos , Mutagênicos/efeitos adversos , Membrana Nuclear/patologia , Epitélio Pigmentado da Retina/patologia , Reparo do DNA , Células Hep G2 , Humanos , Membrana Nuclear/efeitos dos fármacos , Epitélio Pigmentado da Retina/efeitos dos fármacos
7.
Int J Mol Sci ; 22(11)2021 Jun 04.
Artigo em Inglês | MEDLINE | ID: mdl-34199904

RESUMO

Nutrition determines our health, both directly and indirectly. Consumed foods affect the functioning of individual organs as well as entire systems, e.g., the cardiovascular system. There are many different diets, but universal guidelines for proper nutrition are provided in the WHO healthy eating pyramid. According to the latest version, plant products should form the basis of our diet. Many groups of plant compounds with a beneficial effect on human health have been described. Such groups include anthocyanins and iridoids, for which it has been proven that their consumption may lead to, inter alia, antioxidant, cholesterol and lipid-lowering, anti-obesity and anti-diabetic effects. Transcription factors directly affect a number of parameters of cell functions and cellular metabolism. In the context of lipid and cholesterol metabolism, five particularly important transcription factors can be distinguished: liver X receptor (LXR), peroxisome proliferator-activated receptor-α (PPAR-α), peroxisome proliferator-activated receptor-γ (PPAR-γ), CCAAT/enhancer binding protein α (C/EBPα) and sterol regulatory element-binding protein 1c (SREBP-1c). Both anthocyanins and iridoids may alter the expression of these transcription factors. The aim of this review is to collect and systematize knowledge about the impact of anthocyanins and iridoids on transcription factors crucial for lipid and cholesterol homeostasis.


Assuntos
Antocianinas/farmacologia , Colesterol/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Homeostase , Iridoides/farmacologia , Lipídeos/análise , Fatores de Transcrição/metabolismo , Animais , Humanos , Metabolismo dos Lipídeos , Fatores de Transcrição/genética
8.
Nat Commun ; 12(1): 4343, 2021 07 15.
Artigo em Inglês | MEDLINE | ID: mdl-34267224

RESUMO

Aberrant sterol lipid metabolism is associated with physiological dysfunctions in the aging brain and aging-dependent disorders such as neurodegenerative diseases. There is an unmet demand to comprehensively profile sterol lipids spatially and temporally in different brain regions during aging. Here, we develop an ion mobility-mass spectrometry based four-dimensional sterolomics technology leveraged by a machine learning-empowered high-coverage library (>2000 sterol lipids) for accurate identification. We apply this four-dimensional technology to profile the spatially resolved landscapes of sterol lipids in ten functional regions of the mouse brain, and quantitatively uncover ~200 sterol lipids uniquely distributed in specific regions with concentrations spanning up to 8 orders of magnitude. Further spatial analysis pinpoints age-associated differences in region-specific sterol lipid metabolism, revealing changes in the numbers of altered sterol lipids, concentration variations, and age-dependent coregulation networks. These findings will contribute to our understanding of abnormal sterol lipid metabolism and its role in brain diseases.


Assuntos
Química Encefálica , Encéfalo/metabolismo , Lipídeos/química , Esteróis/análise , Envelhecimento/fisiologia , Animais , Feminino , Isomerismo , Lipidômica/métodos , Lipídeos/análise , Aprendizado de Máquina , Camundongos Endogâmicos C57BL , Esteróis/química , Esteróis/metabolismo , Espectrometria de Massas em Tandem/métodos
9.
Methods Mol Biol ; 2314: 109-150, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34235650

RESUMO

The very high content of structurally diverse and biologically active lipids of exotic structures is the hallmark of Mycobacteria. As such the lipid composition is commonly used to characterize mycobacterial strains at the species and type-species levels. The present chapter describes the methods that allow the purification of the most commonly isolated biologically active lipids and those used for analyzing extractable lipids and their constituents, cell wall-linked mycolic acids (MA), and lipoarabinomannan (LAM). These involve various chromatographic techniques and analytical procedures necessary for structural and metabolic studies of mycobacterial lipids. In addition, as the use of physical methods has brought important overhang on chemical structures of the very-long-chain MA, which typify mycobacteria, NMR and mass spectrometry data of these specific fatty acids are included.


Assuntos
Parede Celular/metabolismo , Lipídeos/análise , Lipídeos/isolamento & purificação , Lipopolissacarídeos/análise , Lipopolissacarídeos/isolamento & purificação , Mycobacterium/metabolismo , Espectroscopia de Ressonância Magnética , Espectrometria de Massas
10.
Methods Mol Biol ; 2314: 549-577, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34235670

RESUMO

Decades of study have highlighted the richness and uniqueness of the repertoire of lipid and glycolipid families produced by mycobacteria. Many of these families potently regulate host immune responses, in stimulatory or suppressive ways. Thus, the global study of this repertoire in different genetic backgrounds or under model conditions of infection is gaining interest. Despite the difficulties associated with the specificities of this repertoire, the field of mass spectrometry-based lipidomics of mycobacteria has recently made considerable progress, particularly at the analytical level. There is still considerable scope for further progress, especially with regard to the development of an efficient bioinfomatics pipeline for the analysis of the large datasets generated. This chapter describes an HPLC-MS methodology allowing the simultaneous screening of more than 20 of the lipid families produced by mycobacteria and provides recommendations to analyze the generated data given the state-of-the-art.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Glicolipídeos/metabolismo , Lipidômica/métodos , Lipídeos/análise , Espectrometria de Massas/métodos , Mycobacterium/metabolismo , Glicolipídeos/análise , Humanos
11.
Int J Mol Sci ; 22(13)2021 Jun 23.
Artigo em Inglês | MEDLINE | ID: mdl-34201486

RESUMO

Oleaginous filamentous fungi can accumulate large amount of cellular lipids and biopolymers and pigments and potentially serve as a major source of biochemicals for food, feed, chemical, pharmaceutical, and transport industries. We assessed suitability of Fourier transform (FT) Raman spectroscopy for screening and process monitoring of filamentous fungi in biotechnology. Six Mucoromycota strains were cultivated in microbioreactors under six growth conditions (three phosphate concentrations in the presence and absence of calcium). FT-Raman and FT-infrared (FTIR) spectroscopic data was assessed in respect to reference analyses of lipids, phosphorus, and carotenoids by using principal component analysis (PCA), multiblock or consensus PCA, partial least square regression (PLSR), and analysis of spectral variation due to different design factors by an ANOVA model. All main chemical biomass constituents were detected by FT-Raman spectroscopy, including lipids, proteins, cell wall carbohydrates, and polyphosphates, and carotenoids. FT-Raman spectra clearly show the effect of growth conditions on fungal biomass. PLSR models with high coefficients of determination (0.83-0.94) and low error (approximately 8%) for quantitative determination of total lipids, phosphates, and carotenoids were established. FT-Raman spectroscopy showed great potential for chemical analysis of biomass of oleaginous filamentous fungi. The study demonstrates that FT-Raman and FTIR spectroscopies provide complementary information on main fungal biomass constituents.


Assuntos
Fungos/química , Análise Espectral Raman/métodos , Biomassa , Biotecnologia , Cálcio/metabolismo , Carotenoides/análise , Cromatografia Gasosa , Cromatografia Líquida de Alta Pressão , Análise de Fourier , Fungos/crescimento & desenvolvimento , Lipídeos/análise , Espectroscopia de Ressonância Magnética , Fósforo/análise , Fósforo/metabolismo , Pigmentos Biológicos/análise , Análise de Componente Principal , Espectrofotometria Ultravioleta , Espectroscopia de Infravermelho com Transformada de Fourier
12.
Food Chem ; 362: 130066, 2021 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-34098434

RESUMO

Cereal germ (CG), a by-product of grain milling, has drawn much attention in the food industry because of its nutritional and functional advantages. Nowadays, the utilization of cereal germ from animal feeds to foodstuff is a popular trend. CGs have high content of polyunsaturated fatty acids in their lipids (43.9-64.9% of total fatty acids), but they are also induced to oxidative rancidity under the catalytic reaction of enzymes. Chemical and structural properties of lipids in CGs are affected by different treatments. Thermal and non-thermal effects prevent lipid oxidation or promote lipid combination with starch/protein in CG. Thus, the functional properties and final quality of CG are directly changed. In this review, the chemical composition and application of CGs especially the endogenous lipids are summarized and the effects of various processes on CG lipids/matrices are discussed for CG future development.


Assuntos
Grão Comestível/química , Lipídeos/química , Ração Animal , Animais , Análise de Alimentos , Indústria Alimentícia , Indústria de Processamento de Alimentos , Lipídeos/análise , Oxirredução , Amido/química
13.
Int J Mol Sci ; 22(11)2021 May 24.
Artigo em Inglês | MEDLINE | ID: mdl-34073737

RESUMO

Gestational diabetes mellitus (GDM) is the fastest growing type of diabetes, affecting between 2 to 38% of pregnancies worldwide, varying considerably depending on diagnostic criteria used and sample population studied. Adverse obstetric outcomes include an increased risk of macrosomia, and higher rates of stillbirth, instrumental delivery, and birth trauma. Metabolomics, which is a platform used to analyse and characterise a large number of metabolites, is increasingly used to explore the pathophysiology of cardiometabolic conditions such as GDM. This review aims to summarise metabolomics studies in GDM (from inception to January 2021) in order to highlight prospective biomarkers for diagnosis, and to better understand the dysfunctional metabolic pathways underlying the condition. We found that the most commonly deranged pathways in GDM include amino acids (glutathione, alanine, valine, and serine), carbohydrates (2-hydroxybutyrate and 1,5-anhydroglucitol), and lipids (phosphatidylcholines and lysophosphatidylcholines). We also highlight the possibility of using certain metabolites as predictive markers for developing GDM, with the use of highly stratified modelling techniques. Limitations for metabolomic research are evaluated, and future directions for the field are suggested to aid in the integration of these findings into clinical practice.


Assuntos
Diabetes Gestacional/metabolismo , Redes e Vias Metabólicas , Metabolômica , Aminoácidos/análise , Biomarcadores/análise , Carboidratos/análise , Diabetes Gestacional/sangue , Diabetes Gestacional/diagnóstico , Diabetes Gestacional/urina , Feminino , Humanos , Lipídeos/análise , Espectrometria de Massas , Gravidez
14.
Int J Mol Sci ; 22(10)2021 May 11.
Artigo em Inglês | MEDLINE | ID: mdl-34064761

RESUMO

Kirsten rat sarcoma viral oncogene homolog (KRAS)-driven pancreatic cancer is very lethal, with a five-year survival rate of <9%, irrespective of therapeutic advances. Different treatment modalities including chemotherapy, radiotherapy, and immunotherapy demonstrated only marginal efficacies because of pancreatic tumor specificities. Surgery at the early stage of the disease remains the only curative option, although only in 20% of patients with early stage disease. Clinical trials targeting the main oncogenic driver, KRAS, have largely been unsuccessful. Recently, global metabolic reprogramming has been identified in patients with pancreatic cancer and oncogenic KRAS mouse models. The newly reprogrammed metabolic pathways and oncometabolites affect the tumorigenic environment. The development of methods modulating metabolic reprogramming in pancreatic cancer cells might constitute a new approach to its therapy. In this review, we describe the major metabolic pathways providing acetyl-CoA and NADPH essential to sustain lipid synthesis and cell proliferation in pancreatic cancer cells.


Assuntos
Citratos/metabolismo , Glutamina/metabolismo , Lipídeos/análise , Mitocôndrias/metabolismo , Pâncreas/metabolismo , Neoplasias Pancreáticas/metabolismo , Proteínas Proto-Oncogênicas p21(ras)/metabolismo , Humanos , Pâncreas/citologia , Neoplasias Pancreáticas/patologia , Proteínas Proto-Oncogênicas p21(ras)/genética
15.
Sci Total Environ ; 784: 147018, 2021 Aug 25.
Artigo em Inglês | MEDLINE | ID: mdl-34088028

RESUMO

Fine particulate matter (PM2.5) is able to pass through the respiratory barrier to enter the circulatory system and can consequently spread to the whole body to cause toxicity. Although our previous studies have revealed significantly altered levels of phosphorylcholine-containing lipids in the lungs of rats after chronic inhalation exposure to PM2.5, the effects of PM2.5 on phosphorylcholine-containing lipids in the extrapulmonary organs have not yet been elucidated. In this study, we examined the lipid effects of chronic PM2.5 exposure on various organs and serum by using a rat inhalation model followed by a mass spectrometry-based lipidomic approach. Male Sprague-Dawley rats were continuously exposed at the whole body level to nonfiltered and nonconcentrated ambient air from the outside environment of Taipei city for 8 months, while the control rats inhaled filtered air simultaneously. After exposure, serum samples and various organs, including the testis, pancreas, heart, liver, kidney, spleen, and epididymis, were collected for lipid extraction and analysis to examine the changes in phosphorylcholine-containing lipids after exposure. The results from the partial least squares discriminant analysis models demonstrated that the lipid profiles in the PM2.5 exposure group were different from those in the control group in the rat testis, pancreas, heart, liver, kidney and serum. The greatest PM2.5-induced lipid effects were observed in the testes. Decreased lyso-phosphatidylcholines (PCs) as well as increased unsaturated diacyl-PCs and sphingomyelins in the testes may be related to maintaining the membrane integrity of spermatozoa, antioxidation, and cell signaling. Additionally, our results showed that decreased PC(16:0/18:1) was observed in both the serum and testes. In conclusion, exposure to chronic environmental concentrations of PM2.5 caused lipid perturbation, especially in the testes of rats. This study highlighted the susceptibility of the testes and suggested possible molecular events for future study.


Assuntos
Poluentes Atmosféricos , Material Particulado , Poluentes Atmosféricos/análise , Poluentes Atmosféricos/toxicidade , Animais , Cidades , Lipídeos/análise , Masculino , Material Particulado/análise , Material Particulado/toxicidade , Ratos , Ratos Sprague-Dawley
16.
Int J Mol Sci ; 22(11)2021 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-34071405

RESUMO

In vitro skin tissue engineering is challenging due to the manifold differences between the in vivo and in vitro conditions. Yet, three-dimensional (3D) human skin equivalents (HSEs) are able to mimic native human skin in many fundamental aspects. However, the epidermal lipid barrier formation, which is essential for the functionality of the skin barrier, remains compromised. Recently, HSEs with an improved lipid barrier formation were generated by (i) incorporating chitosan in the dermal collagen matrix, (ii) reducing the external oxygen level to 3%, and (iii) inhibiting the liver X receptor (LXR). In this study, we aimed to determine the synergic effects in full-thickness models (FTMs) with combinations of these factors as single-, double-, and triple-targeted optimization approaches. The collagen-chitosan FTM supplemented with the LXR inhibitor showed improved epidermal morphogenesis, an enhanced lipid composition, and a better lipid organization. Importantly, barrier functionality was improved in the corresponding approach. In conclusion, our leading optimization approach substantially improved the epidermal morphogenesis, barrier formation, and functionality in the FTM, which therefore better resembled native human skin.


Assuntos
Células Epidérmicas/metabolismo , Epiderme/metabolismo , Morfogênese , Pele/metabolismo , Engenharia Tecidual/métodos , Células Cultivadas , Quitosana/metabolismo , Cromatografia Líquida , Colágeno/metabolismo , Epiderme/crescimento & desenvolvimento , Humanos , Queratinócitos/citologia , Queratinócitos/metabolismo , Metabolismo dos Lipídeos , Lipídeos/análise , Receptores X do Fígado/metabolismo , Espectrometria de Massas , Espalhamento a Baixo Ângulo , Pele/citologia , Pele/crescimento & desenvolvimento , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios X
17.
Anal Bioanal Chem ; 413(16): 4181-4194, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33974088

RESUMO

This work assesses the potential of new water cluster-based ion beams for improving the capabilities of secondary ion mass spectrometry (SIMS) for in situ lipidomics. The effect of water clusters was compared to carbon dioxide clusters, along with the effect of using pure water clusters compared to mixed water and carbon dioxide clusters. A signal increase was found when using pure water clusters. However, when analyzing cells, a more substantial signal increase was found in positive ion mode when the water clusters also contained carbon dioxide, suggesting that additional reactions are in play. The effects of using a water primary ion beam on a more complex sample were investigated by analyzing brain tissue from an Alzheimer's disease transgenic mouse model. The results indicate that the ToF-SIMS results are approaching those from MALDI as ToF-SIMS was able to image lyso-phosphocholine (LPC) lipids, a lipid class that for a long time has eluded detection during SIMS analyses. Gangliosides, sulfatides, and cholesterol were also imaged.


Assuntos
Doença de Alzheimer/patologia , Encéfalo/patologia , Dióxido de Carbono/análise , Lipídeos/análise , Água/análise , Animais , Química Encefálica , Modelos Animais de Doenças , Humanos , Células MCF-7 , Camundongos , Camundongos Transgênicos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Espectrometria de Massa de Íon Secundário
18.
J Steroid Biochem Mol Biol ; 211: 105903, 2021 07.
Artigo em Inglês | MEDLINE | ID: mdl-33933575

RESUMO

We aimed to evaluate sex differences in changes of lipid profiles in a cohort of metabolically healthy adults following Orthodox fasting (OF), as well as to assess a potential role of vitamin D status in mediating these variations. 45 individuals (24 premenopausal females, 53.3 %) with mean age 48.3 ± 9.1 years and mean Body Mass Index 28.7 ± 5.8 kg/m2 were prospectively followed for 12 weeks. Anthropometry, dietary and biochemical data regarding serum lipids, and vitamin D status were collected at baseline, 7 weeks after the implementation of OF, and 5 weeks after fasters returned to their standard dietary habits (12 weeks from baseline). According to 25-hydroxy-vitamin D [25(OH)D] measurements, participants were divided into two groups: those with concentrations above and below the median of values. Females with 25(OH)D concentrations below the median manifested a non-significant reduction by approximately 15 % in total and low-density lipoprotein cholesterol during the fasting period, followed by a significant increase 5 weeks after OF cessation (170.7 vs. 197.5 and 99.6 vs. 121.0 mg/dl respectively, p < 0.001). In contrast, males with 25(OH)D levels below the median demonstrated an inverse, non-significant trend of increase in lipid concentrations during the whole study period. Our findings suggest strikingly different inter-gender lipid responses to a dietary model of low-fat, mediated by vitamin D status. Further studies are necessary to reveal the underlying mechanisms and assess the importance of these differences with respect to cardiovascular health and the benefit of vitamin D supplementation strategies.


Assuntos
Índice de Massa Corporal , Suplementos Nutricionais , Jejum , Lipídeos/análise , Vitamina D/metabolismo , Vitaminas/metabolismo , Antropometria , Ortodoxia Oriental , Comportamento Alimentar , Feminino , Grécia , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Fatores Sexuais
19.
Methods Mol Biol ; 2295: 3-13, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34047968

RESUMO

Analysis of plant lipids provides insights into a range of biological processes, from photosynthetic membrane function to oil seed engineering. Many lipid extraction protocols are tailored to fit a specific lipid class. Here we describe a procedure for extraction of glycerolipids from vegetative tissue. This procedure is designed for 1 gram of tissue per sample but maybe scaled for larger samples.


Assuntos
Lipídeos/isolamento & purificação , Extração Líquido-Líquido/métodos , Plantas/metabolismo , Clorofórmio/química , Glicerol/metabolismo , Lipídeos/análise , Metanol/química , Sementes/química , Solventes/química , Água/química
20.
Clin Transl Gastroenterol ; 12(3): e00316, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33939383

RESUMO

INTRODUCTION: This randomized trial aimed to compare the effects of moderate-intensity aerobic vs resistance exercise with dietary modification in patients with nonalcoholic fatty liver disease (NAFLD). METHODS: Patients with NAFLD were randomly assigned (1:1) to a 12-week supervised training program of moderate-intensity aerobic or resistance exercise with dietary intervention consisting of monthly individual nutritional counseling by a dietician. Transient elastography, anthropometry, body composition, cardiorespiratory fitness, biochemistries, and glucose tolerance were measured at baseline and 12 weeks. RESULTS: Eighteen subjects exercised for an average of 3.35 ± 0.30 sessions a week in the aerobic group, and 17 subjects exercised an average of 3.39 ± 0.28 sessions a week in the resistance group. After completion of the training program, hepatic fat content was similarly reduced in both groups (P < 0.001). The mean relative reduction from baseline in the aerobic group was -10.3% (95% confidence interval -18.2 to -2.40) and the resistance group was -12.6% (-20.5 to -4.69). Liver steatosis (defined as controlled attenuation parameter >248 dB/m) disappeared in 9 (50%) of the aerobic group and in 9 (53%) of the resistance group. Whole-body and muscle insulin sensitivity indexes were improved, and waist circumference was reduced comparably in both exercise groups. The number of exercise sessions per week was correlated with the absolute reduction in hepatic fat content (r = 0.52; P = 0.001). Weekly exercise training ≥3 sessions substantially attenuates liver fat accumulation independent of weight loss. DISCUSSION: Moderate-intensity aerobic training and resistance training with dietary modification are equally effective for reducing intrahepatic fat and improving underlying insulin resistance among patients with NAFLD.


Assuntos
Aconselhamento/métodos , Dietética/métodos , Hepatopatia Gordurosa não Alcoólica/terapia , Treinamento de Força/métodos , Adulto , Aptidão Cardiorrespiratória/fisiologia , Terapia Combinada/métodos , Técnicas de Imagem por Elasticidade , Feminino , Humanos , Metabolismo dos Lipídeos/fisiologia , Lipidômica , Lipídeos/análise , Fígado/diagnóstico por imagem , Fígado/patologia , Masculino , Hepatopatia Gordurosa não Alcoólica/diagnóstico , Nutricionistas , Resultado do Tratamento
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