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1.
Int J Mol Sci ; 22(23)2021 Nov 23.
Artigo em Inglês | MEDLINE | ID: mdl-34884471

RESUMO

Obesity is a major health problem. Compelling evidence supports the beneficial effects of probiotics on obesity. However, the anti-obesity effect of probiotics remains unknown. In this study, we investigated the anti-obesity effects and potential mechanisms of Lactiplantibacillus plantarum ATG-K2 using 3T3-L1 adipocytes and high-fat diet (HFD)-induced obese mice. 3T3-L1 cells were incubated to determine the effect of lipid accumulation with lysate of L. plantarum ATG-K2. Mice were fed a normal fat diet or HFD with L. plantarum ATG-K2 and Orlistat for 8 weeks. L. plantarum ATG-K2 inhibited lipid accumulation in 3T3-L1 adipocytes, and reduced body weight gain, WAT weight, and adipocyte size in HFD-induced obese mice, concurrently with the downregulation of PPARγ, SREBP1c, and FAS and upregulation of PPARα, CTP1, UCP1, Prdm16, and ND5. Moreover, L. plantarum ATG-K2 decreased TG, T-CHO, leptin, and TNF-α levels in the serum, with corresponding gene expression levels in the intestine. L. plantarum ATG-K2 modulated the gut microbiome by increasing the abundance of the Lactobacillaceae family, which increased SCFA levels and branched SCFAs in the feces. L. plantarum ATG-K2 exhibited an anti-obesity effect and anti-hyperlipidemic effect in 3T3-L1 adipocytes and HFD-induced obese mice by alleviating the inflammatory response and regulating lipid metabolism, which may be influenced by modulation of the gut microbiome and its metabolites. Therefore, L. plantarum ATG-K2 can be a preventive and therapeutic agent for obesity.


Assuntos
Dieta Hiperlipídica/efeitos adversos , Lactobacillaceae/fisiologia , Obesidade/dietoterapia , Probióticos/administração & dosagem , Células 3T3-L1 , Animais , Fatores Biológicos/análise , Peso Corporal , Modelos Animais de Doenças , Microbioma Gastrointestinal/efeitos dos fármacos , Regulação da Expressão Gênica , Lactobacillaceae/química , Camundongos , Camundongos Obesos , Obesidade/induzido quimicamente , Obesidade/genética , Probióticos/farmacologia
2.
J Bacteriol ; 201(5)2019 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-30530517

RESUMO

Pseudomonas aeruginosa is an opportunistic pathogen found ubiquitously in the environment and commonly associated with airway infection in patients with cystic fibrosis. P. aeruginosa strain PAO1 is one of the most commonly used laboratory-adapted research strains and is a standard laboratory-adapted strain in multiple laboratories and strain banks worldwide. Due to potential isolate-to-isolate variability, we investigated the genomic and phenotypic diversity among 10 PAO1 strains (henceforth called sublines) obtained from multiple research laboratories and commercial sources. Genomic analysis predicted a total of 5,682 genes, with 5,434 (95.63%) being identical across all 10 strains. Phenotypic analyses revealed comparable growth phenotypes in rich media and biofilm formation profiles. Limited differences were observed in antibiotic susceptibility profiles and immunostimulatory potential, measured using heat-killed whole-cell preparations in four immortalized cell lines followed by quantification of interleukin-6 (IL-6) and IL-1ß secretion. However, variability was observed in the profiles of secreted molecular products, most notably, in rhamnolipid, pyoverdine, pyocyanin, Pseudomonas quinolone signal (PQS), extracellular DNA, exopolysaccharide, and outer membrane vesicle production. Many of the observed phenotypic differences did not correlate with subline-specific genetic changes, suggesting alterations in transcriptional and translational regulation. Taken together, these results suggest that individually maintained sublines of PAO1, even when acquired from the same parent subline, are continuously undergoing microevolution during culture and storage that results in alterations in phenotype, potentially affecting the outcomes of in vitro phenotypic analyses and in vivo pathogenesis studies.IMPORTANCE Laboratory-adapted strains of bacteria are used throughout the world for microbiology research. These prototype strains help keep research data consistent and comparable between laboratories. However, we have observed phenotypic variability when using different strains of Pseudomonas aeruginosa PAO1, one of the major laboratory-adopted research strains. Here, we describe the genomic and phenotypic differences among 10 PAO1 strains acquired from independent sources over 15 years to understand how individual maintenance affects strain characteristics. We observed limited genomic changes but variable phenotypic changes, which may have consequences for cross-comparison of data generated using different PAO1 strains. Our research highlights the importance of limiting practices that may promote the microevolution of model strains and calls for researchers to specify the strain origin to ensure reproducibility.


Assuntos
Fatores Biológicos/análise , Variação Genética , Genômica , Pseudomonas aeruginosa/genética , Pseudomonas aeruginosa/fisiologia , Biofilmes/crescimento & desenvolvimento , Meios de Cultura/química , Citocinas/metabolismo , Evolução Molecular , Genótipo , Testes de Sensibilidade Microbiana , Fenótipo , Pseudomonas aeruginosa/imunologia , Seleção Genética
3.
Biotechnol Bioeng ; 116(1): 121-131, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30199089

RESUMO

Large-scale manufacturing of therapeutic cells requires bioreactor technologies with online feedback control enabled by monitoring of secreted biomolecular critical quality attributes (CQAs). Electrospray ionization mass spectrometry (ESI-MS) is a highly sensitive label-free method to detect and identify biomolecules, but requires extensive sample preparation before analysis, making online application of ESI-MS challenging. We present a microfabricated, monolithically integrated device capable of continuous sample collection, treatment, and direct infusion for ESI-MS detection of biomolecules in high-salt solutions. The dynamic mass spectrometry probe (DMSP) uses a microfluidic mass exchanger to rapidly condition samples for online MS analysis by removing interfering salts, while concurrently introducing MS signal enhancers to the sample for sensitive biomolecular detection. Exploiting this active conditioning capability increases MS signal intensity and signal-to-noise ratio. As a result, sensitivity for low-concentration biomolecules is significantly improved, and multiple proteins can be detected from chemically complex samples. Thus, the DMSP has significant potential to serve as an enabling portion of a novel analytical tool for discovery and monitoring of CQAs relevant to therapeutic cell manufacturing.


Assuntos
Fatores Biológicos/análise , Reatores Biológicos , Técnicas de Cultura de Células/métodos , Espectrometria de Massas por Ionização por Electrospray/métodos , Tecnologia Farmacêutica/métodos , Terapia Baseada em Transplante de Células e Tecidos
4.
Appl Microbiol Biotechnol ; 103(16): 6759-6769, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31230100

RESUMO

Confocal Raman microspectral imaging (CRMI) is an advanced cell-imaging method that maps endogenous molecular compositions with their unique spectral fingerprint indicators. The aim of this work was to provide a visualized understanding of subcellular features of live osteosarcoma cells using a 532-nm laser excitation without the use of dyes or molecular probes. Both malignant osteoblast and spindle osteosarcoma cells derived from the BALB/c mouse osteosarcoma cell line K7M2 were investigated in this work. After preprocessing the obtained spectral dataset, K-means cluster analysis (KCA) is employed to reconstruct Raman spectroscopic maps of single biological cells by identifying regions of the cellular membrane, cytoplasm, organelles, and nucleus with their corresponding mean spectra. Principal component analysis (PCA) was further employed to indicate variables of significant influence on the separation of the spectra of each cellular component. The biochemical components of the two cell types were then extracted by showing the spectral and distribution features attributed to proteins, lipids, and DNA. Using this standardized CRMI technique and multivariate analysis approaches, the results obtained could be a sound foundation for a typical Raman imaging protocol of live cellular biomedical analysis.


Assuntos
Fatores Biológicos/análise , Linhagem Celular Tumoral/química , Microscopia Óptica não Linear/métodos , Osteossarcoma/patologia , Análise de Célula Única/métodos , Animais , Camundongos , Camundongos Endogâmicos BALB C , Análise Multivariada
5.
J Microsc ; 270(1): 83-91, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29064560

RESUMO

The symbiotic relationship between corals and Symbiodinium spp. is the key to the success and survival of coral reef ecosystems the world over. Nutrient exchange and chemical communication between the two partners provides the foundation of this key relationship, yet we are far from a complete understanding of these processes. This is due, in part, to the difficulties associated with studying an intracellular symbiosis at the small spatial scales required to elucidate metabolic interactions between the two partners. This feasibility study, which accompanied a more extensive investigation of fixed Symbiodinium cells (data unpublished), examines the potential of using synchrotron radiation infrared microspectroscopy (SR-IRM) for exploring metabolite localisation within a single Symbiodinium cell. In doing so, three chemically distinct subcellular regions of a single Symbiodinium cell were established and correlated to cellular function based on assignment of diagnostic chemical classes.


Assuntos
Fatores Biológicos/análise , Dinoflagellida/química , Dinoflagellida/ultraestrutura , Microscopia/métodos , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Estudos de Viabilidade
6.
Curr Microbiol ; 75(2): 142-149, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-28918535

RESUMO

Marine actinomycetes are less investigated compared to terrestrial strains as potential sources of natural products. To date, few investigations have been performed on culturable actinomycetes associated with South China Sea sediments. In the present study, twenty-eight actinomycetes were recovered from South China Sea sediments after dereplication by traditional culture-dependent method. The 16S rRNA gene sequences analyses revealed that these strains related to five families and seven genera. Twelve representative strains possessed at least one of the biosynthetic genes coding for polyketide synthase I, II, and nonribosomal peptide synthetase. Four strains had anti-Mycobacterium phlei activities and five strains had activities against methicillin-resistant Staphylococcus aureus. 10 L-scale fermentation of strains Salinispora sp. NHF45, Nocardiopsis sp. NHF48, and Streptomyces sp. NHF86 were carried out for novel and bioactive compounds discovery. Finally, we obtained a novel α-pyrone compound from marine Nocardiopsis sp. NHF48, an analogue of paulomenol from marine Streptomyces sp. NHF86 and a new source of rifamycin B, produced by Salinispora sp. NHF45. The present study concluded that marine actinomycetes, which we isolated from South China Sea sediments, will be a suitable source for the development of novel and bioactive compounds.


Assuntos
Actinobacteria/genética , Actinobacteria/isolamento & purificação , Organismos Aquáticos/genética , Organismos Aquáticos/isolamento & purificação , Fatores Biológicos/análise , Bioprospecção , Sedimentos Geológicos/microbiologia , Actinobacteria/classificação , Actinobacteria/metabolismo , Organismos Aquáticos/classificação , Organismos Aquáticos/metabolismo , China , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Filogenia , RNA Ribossômico 16S/genética , Análise de Sequência de DNA
7.
Anal Chem ; 89(7): 4062-4070, 2017 04 04.
Artigo em Inglês | MEDLINE | ID: mdl-28263059

RESUMO

Some pathogens and toxins have the potential to be used as weapons of mass destruction and instigate population-based fear. Rapid, sensitive, and unambiguous identification of biothreat agents is of paramount importance for confirmation of the event and to mitigate the direct and indirect damages to public health and resources. Although there are several potential dissemination scenarios to describe an attack with a biological weapon, artificially generated bioaerosol is of the greatest concern from a bioterrorism or warfare perspective, potentially capable of causing mass destruction to a civilian or military population by inhalation of toxic bioaerosol. The present investigation proposes methodologies for recovery of biological agent followed by an off-site unambiguous detection using tandem mass spectrometry, in a postattack situation. We envisaged a biothreat scenario wherein the polydispersed bioaerosol is disseminated in bulk over any geographical setting. The larger particles (>5 µm in diameter) of bioaerosol settle and bind to various surfaces depending on the geographical setting. Recovery of agent was optimized from foliage, sand, and glass in a simulated biothreat scenario using bovine serum albumin (BSA). The recovered agents were shown to be amenable to detection by a downstream tandem MS analysis. Applicability of the proposed methodology was demonstrated in validation experiments for the recovery and detection of toxin and bacterial agents. The use of cleaner matrices (foliage, exposed smooth surfaces, sand) is recommended for retrospective verification of agent in a biothreat scenario.


Assuntos
Fatores Biológicos/análise , Armas Biológicas , Animais , Bovinos , Clostridium perfringens/química , Soroalbumina Bovina/química , Espectrometria de Massas em Tandem
8.
J Nutr ; 147(6): 1121-1130, 2017 06.
Artigo em Inglês | MEDLINE | ID: mdl-28298536

RESUMO

Background: Holder pasteurization (HP) destroys multiple bioactive factors in donor human milk (DM), and UV-C irradiation (UVC) is potentially a gentler method for pasteurizing DM for preterm infants.Objective: We investigated whether UVC-treated DM improves gut maturation and resistance toward bacterial infections relative to HP-treated DM.Methods: Bacteria, selected bioactive components, and markers of antioxidant capacity were measured in unpasteurized donor milk (UP), HP-treated milk, and UVC-treated milk (all from the same DM pool). Fifty-seven cesarean-delivered preterm pigs (91% gestation; ratio of males to females, 30:27) received decreasing volumes of parental nutrition (average 69 mL · kg-1 · d-1) and increasing volumes of the 3 DM diets (n = 19 each, average 89 mL · kg-1 · d-1) for 8-9 d. Body growth, gut structure and function, and systemic bacterial infection were evaluated.Results: A high bacterial load in the UP (6×105 colony forming units/mL) was eliminated similarly by HP and UVC treatments. Relative to HP-treated milk, both UVC-treated milk and UP showed greater activities of lipase and alkaline phosphatase and concentrations of lactoferrin, secretory immunoglobulin A, xanthine dehydrogenase, and some antioxidant markers (all P < 0.05). The pigs fed UVC-treated milk and pigs fed UP showed higher relative weight gain than pigs fed HP-treated milk (5.4% and 3.5%), and fewer pigs fed UVC-treated milk had positive bacterial cultures in the bone marrow (28%) than pigs fed HP-treated milk (68%) (P < 0.05). Intestinal health was also improved in pigs fed UVC-treated milk compared with those fed HP-treated milk as indicated by a higher plasma citrulline concentration (36%) and villus height (38%) (P < 0.05) and a tendency for higher aminopeptidase N (48%) and claudin-4 (26%) concentrations in the distal intestine (P < 0.08). The gut microbiota composition was similar among groups except for greater proportions of Enterococcus in pigs fed UVC-treated milk than in pigs fed UP and those fed HP-treated milk in both cecum contents (20% and 10%) and distal intestinal mucosa (24% and 20%) (all P < 0.05).Conclusions: UVC is better than HP treatment in preserving bioactive factors in DM. UVC-treated milk may induce better weight gain, intestinal health, and resistance against bacterial infections as shown in preterm pigs as a model for DM-fed preterm infants.


Assuntos
Infecções Bacterianas/prevenção & controle , Dieta , Irradiação de Alimentos/métodos , Idade Gestacional , Intestinos/crescimento & desenvolvimento , Leite Humano/efeitos da radiação , Aumento de Peso , Animais , Animais Recém-Nascidos , Antioxidantes/metabolismo , Fatores Biológicos/análise , Medula Óssea/microbiologia , Enterococcus/crescimento & desenvolvimento , Feminino , Microbioma Gastrointestinal , Humanos , Imunoglobulina A Secretora/análise , Recém-Nascido , Mucosa Intestinal/metabolismo , Mucosa Intestinal/microbiologia , Intestinos/microbiologia , Masculino , Leite Humano/química , Leite Humano/enzimologia , Pasteurização/métodos , Suínos , Raios Ultravioleta
9.
Virol J ; 14(1): 144, 2017 07 27.
Artigo em Inglês | MEDLINE | ID: mdl-28750646

RESUMO

BACKGROUND: Echovirus 30 (E30) causes acute aseptic meningitis. Viral replication requires energy and macromolecular precursors derived from the metabolic network of the host cell. The effect of viral infection within a host cell metabolic activity remains unclear. METHODS: To gain an insight into cell-virus interaction during E30 infection we used a human rhabdomyosarcoma cell line. In a new approach to metabolomics, 1H NMR was used to measure the level of various cellular metabolites at different times of infection and morphological examination of the cells. Statistical analysis was done by using Confidence interval (CI) 95% and One-way ANOVA test. RESULTS: The1H NMR metabolite spectrum signals were observed between mock infected and virus infected cells. Both mock infected and virus infected cells utilized glucose through metabolic pathways and released metabolic end products. Upon infection, the concentration of Alanine, Lactate, Acetate, Glutamate, Tyrosine, Histidine, Phenylalanine, Creatine, Choline and Formate, increased. Interestingly, all of these augmented metabolites were decreased during later stage of infection. The cells showed wide-ranging lipid signals at the end of infection, which correlates with the morphological changes as apoptosis (programmed cell death) of cells was observed. A significant association was found between time interval (12 h, 24 h, and 48 h) and metabolites likewise Alanin, Lactate, Acetate, Glutamate, Tyrosine, Histidine, Phenylalanine, Creatine, Choline and Formate respectively released by cell during infection, which is highly significant (p < 0.01). CONCLUSION: Progressive breakdown and utilization of all cellular components were observed as the infection increased. This study is useful for monitoring the cellular metabolic changes during viral infection.


Assuntos
Fatores Biológicos/análise , Enterovirus Humano B/crescimento & desenvolvimento , Metaboloma , Linhagem Celular Tumoral , Humanos , Espectroscopia de Ressonância Magnética
10.
BMC Complement Altern Med ; 17(1): 22, 2017 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-28056926

RESUMO

BACKGROUND: Edible Bird's Nest (EBN) as a popular traditional Chinese medicine is believed to have health enhancing and antiviral activities against influenza A virus (IAV); however, the molecular mechanism behind therapeutic effects of EBN is not well characterized. METHODS: In this study, EBNs that underwent different enzymatic preparation were tested against IAV infected cells. 50% cytotoxic concentration (CC50) and 50% inhibitory concentration (IC50) of the EBNs against IAV strain A/Puerto Rico/8/1934(H1N1) were determined by HA and MTT assays. Subsequently, the sialic acid content of the used EBNs were analyzed by fluorometric HPLC. Western Blotting and immunofluorescent staining were used to investigate the effects of EBNs on early endosomal trafficking and autophagy process of influenza virus. RESULTS: This study showed that post inoculations of EBNs after enzymatic preparations have the highest efficacy to inhibit IAV. While CC50 of the tested EBNs ranged from 27.5-32 mg/ml, the IC50 of these compounds ranged between 2.5-4.9 mg/ml. EBNs could inhibit IAV as efficient as commercial antiviral agents, such as amantadine and oseltamivir with different mechanisms of action against IAV. The antiviral activity of these EBNs correlated with the content of N-acetyl neuraminic acid. EBNs could affect early endosomal trafficking of the virus by reducing Rab5 and RhoA GTPase proteins and also reoriented actin cytoskeleton of IAV infected cells. In addition, for the first time this study showed that EBNs can inhibit intracellular autophagy process of IAV life cycle as evidenced by reduction of LC3-II and increasing of lysosomal degradation. CONCLUSIONS: The results procured in this study support the potential of EBNs as supplementary medication or alternative to antiviral agents to inhibit influenza infections. Evidently, EBNs can be a promising antiviral agent; however, these natural compounds should be screened for their metabolites prior to usage as therapeutic approach.


Assuntos
Antivirais/farmacologia , Fatores Biológicos/farmacologia , Aves , Vírus da Influenza A Subtipo H1N1/efeitos dos fármacos , Vírus da Influenza A/efeitos dos fármacos , Influenza Humana/virologia , Citoesqueleto de Actina/efeitos dos fármacos , Citoesqueleto de Actina/metabolismo , Animais , Antivirais/análise , Antivirais/metabolismo , Fatores Biológicos/análise , Fatores Biológicos/metabolismo , Aves/metabolismo , Humanos , Vírus da Influenza A Subtipo H1N1/fisiologia , Vírus da Influenza A/fisiologia , Influenza Humana/metabolismo , Proteínas rab5 de Ligação ao GTP/metabolismo , Proteína rhoA de Ligação ao GTP/metabolismo
11.
Environ Manage ; 59(4): 652-664, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28091741

RESUMO

Aquatic invertebrates form the base of the consumer food web in lakes. In coal-mining end-pit lakes, invertebrates are exposed to an environment with potentially challenging physical and chemical features. We hypothesized that the physical and chemical features of end-pit lakes reduce critical littoral habitat and thus reduce invertebrate diversity, thereby limiting the potential for these lakes to be naturalized. We used a multivariate approach using principle component analysis and redundancy analysis to study relationships between invertebrate community structure, habitat features, and water quality in five end-pit lakes and five natural lakes in the Rocky Mountain foothills of west-central Alberta, Canada. Results show a significantly different invertebrate community structure was present in end-pit lakes as compared with reference lakes in the same region, which could be accounted for by water hardness, conductivity, slope of the littoral zone, and phosphorus concentrations. Habitat diversity in end-pit lakes was also limited, cover provided by macrophytes was scarce, and basin slopes were significantly steeper in pit lakes. Although water chemistry is currently the strongest influencing factor on the invertebrate community, physical challenges of habitat homogeneity and steep slopes in the littoral zones were identified as major drivers of invertebrate community structure. The addition of floating wetlands to the littoral zone of existing pit lakes can add habitat complexity without the need for large-scale alterations to basing morphology, while impermeable capping of waste-rock and the inclusion of littoral habitat in the planning process of new pit lakes can improve the success of integrating new pit lakes into the landscape.


Assuntos
Fatores Biológicos/análise , Minas de Carvão , Ecossistema , Sedimentos Geológicos/química , Invertebrados/crescimento & desenvolvimento , Lagos/química , Alberta , Animais , Invertebrados/química , Fósforo/análise , Análise de Componente Principal , Qualidade da Água
12.
Opt Express ; 22(7): 8165-89, 2014 Apr 07.
Artigo em Inglês | MEDLINE | ID: mdl-24718194

RESUMO

A system for measuring spectrally-resolved fluorescence cross sections of single bioaerosol particles has been developed and employed in a biological safety level 3 (BSL-3) facility at Edgewood Chemical and Biological Center (ECBC). It is used to aerosolize the slurry or solution of live agents and surrogates into dried micron-size particles, and to measure the fluorescence spectra and sizes of the particles one at a time. Spectrally-resolved fluorescence cross sections were measured for (1) bacterial spores: Bacillus anthracis Ames (BaA), B. atrophaeus var. globigii (BG) (formerly known as Bacillus globigii), B. thuringiensis israelensis (Bti), B. thuringiensis kurstaki (Btk), B. anthracis Sterne (BaS); (2) vegetative bacteria: Escherichia coli (E. coli), Pantoea agglomerans (Eh) (formerly known as Erwinia herbicola), Yersinia rohdei (Yr), Yersinia pestis CO92 (Yp); and (3) virus preparations: Venezuelan equine encephalitis TC83 (VEE) and the bacteriophage MS2. The excitation wavelengths were 266 nm, 273 nm, 280 nm, 365 nm and 405 nm.


Assuntos
Bactérias/química , Fatores Biológicos/análise , Espectrometria de Fluorescência/métodos , Fluorescência
13.
Crit Rev Food Sci Nutr ; 52(9): 761-74, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22698267

RESUMO

It is essential that the novel biomarkers discovered by means of advanced detection tools based on metabonomics could be used for long-term monitoring in food safety. By summarizing the common biomarkers discovery flowsheet based on metabonomics, this review evaluates the possible application of metabonomics in new biomarker discovery, especially in relation to food safety issues. Metabonomics have the advantages of decreasing detection limits and constant monitoring. Although metabonomics is still in the developmental stage, we believe that, based on its properties, such as noninvasiveness, sensitivity, and persistence, together with rigorous experimental designs, new and novel technologies, as well as increasingly accurate chemometrics and a relational database, metabonomics can demonstrate extensive application in food safety in the postgenome period.


Assuntos
Inocuidade dos Alimentos/métodos , Metabolômica/métodos , Animais , Fatores Biológicos/análise , Bases de Dados Factuais , Inspeção de Alimentos/métodos , Inspeção de Alimentos/tendências , Saúde Global , Humanos , Metabolômica/tendências
14.
Acta Neurol Scand ; 125(4): 219-27, 2012 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22070461

RESUMO

Fatigue is a common and disabling consequence of stroke. Its mechanisms are unknown. Neuroanatomical abnormalities (e.g. white matter lesions, brain atrophy), neuroendocrine dysregulation, neurotransmitter changes and inflammation are associated with fatigue in conditions other than stroke. This review sought to identify published studies describing associations between post-stroke fatigue and these biological factors. We searched Medline, EMBASE, CINAHL, PsycINFO and AMED on October 15 and PubMed on 28 December 2010 and included studies in English that recruited at least 10 patients (>18 years old) with stroke, assessed fatigue and reported its relationship with neuroanatomical abnormalities, hypothalamo-pituitary-adrenal axis dysregulation, neurotransmitter changes or inflammation. Of 4916 citations from the searches, 17 studies met our inclusion criteria. There was no association between white matter lesions, brain atrophy or pathological type of stroke and fatigue (seven studies, n = 4746). The data on relationship between lesion location and fatigue were inconclusive: four (n = 675) of 13 studies (n = 1613) showed associations between fatigue and infratentorial lesion location (brainstem in particular) or basal ganglia stroke. One study reported C-reactive protein levels and found an association with fatigue. No studies reported hypothalamo-pituitary-adrenal axis dysregulation or neurotransmitter changes and fatigue. We could not perform meta-analysis because the studies used different methods of fatigue assessment, examined different populations and had different designs. The biological mechanisms of post-stroke fatigue are uncertain. Further studies are required to determine the relationship between post-stroke fatigue and biological factors.


Assuntos
Fadiga/etiologia , Acidente Vascular Cerebral/complicações , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Fatores Biológicos/análise , Encéfalo/patologia , Proteína C-Reativa/metabolismo , Fadiga/metabolismo , Fadiga/patologia , Humanos , Pessoa de Meia-Idade , Neurotransmissores/metabolismo , Adulto Jovem
15.
Bioinformatics ; 26(18): i653-8, 2010 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-20823335

RESUMO

MOTIVATION: Detecting human proteins that are involved in virus entry and replication is facilitated by modern high-throughput RNAi screening technology. However, hit lists from different laboratories have shown only little consistency. This may be caused by not only experimental discrepancies, but also not fully explored possibilities of the data analysis. We wanted to improve reliability of such screens by combining a population analysis of infected cells with an established dye intensity readout. RESULTS: Viral infection is mainly spread by cell-cell contacts and clustering of infected cells can be observed during spreading of the infection in situ and in vivo. We employed this clustering feature to define knockdowns which harm viral infection efficiency of human Hepatitis C Virus. Images of knocked down cells for 719 human kinase genes were analyzed with an established point pattern analysis method (Ripley's K-function) to detect knockdowns in which virally infected cells did not show any clustering and therefore were hindered to spread their infection to their neighboring cells. The results were compared with a statistical analysis using a common intensity readout of the GFP-expressing viruses and a luciferase-based secondary screen yielding five promising host factors which may suit as potential targets for drug therapy. CONCLUSION: We report of an alternative method for high-throughput imaging methods to detect host factors being relevant for the infection efficiency of viruses. The method is generic and has the potential to be used for a large variety of different viruses and treatments being screened by imaging techniques.


Assuntos
Fatores Biológicos/análise , Hepacivirus/fisiologia , Processamento de Imagem Assistida por Computador , Interferência de RNA , RNA Interferente Pequeno , Replicação Viral , Antígenos CD/análise , Caseína Quinase II/análise , Linhagem Celular Tumoral , Interpretação Estatística de Dados , Vírus da Dengue/fisiologia , Humanos , Antígenos de Histocompatibilidade Menor , Fosfotransferases (Aceptor do Grupo Álcool)/análise , Proteínas Quinases/genética , Receptores de Superfície Celular/análise , Membro 1 da Família de Moléculas de Sinalização da Ativação Linfocitária , Tetraspanina 28 , Receptor 3 de Fatores de Crescimento do Endotélio Vascular/análise
16.
BMC Complement Altern Med ; 11: 82, 2011 Sep 26.
Artigo em Inglês | MEDLINE | ID: mdl-21943200

RESUMO

BACKGROUND: Keloid is a type of scar which extends beyond the boundaries of the original wound. It can spread to the surrounding skin by invasion. The use of Tualang honey is a possible approach for keloid treatment. The objective of this study was to determine the antiproliferative effect of methanolic extraction of Tualang honey to primary human keloid fibroblasts and to identify the volatile compounds in methanol extraction of Tualang honey. METHODS: Crude Tualang honey was extracted with methanol and then dried using rota vapor to remove remaining methanol from honey. Normal and keloid fibroblasts were verified and treated with the extracted honey. Cell proliferation was tested with [3-(4,5-dimethylthiazol-2-yi)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt] (MTS) assay. Extraction of Tualang honey using methanol was carried out and the extracted samples were analysed using gas chromatography-mass spectrometry (GC-MS). The result was analysed using SPSS and tested with Kruskal-Wallis and Mann-Whitney tests. RESULTS: Methanolic extraction of honey has positive anti proliferative effect on keloid fibroblasts in a dose-dependent manner. The presence of fatty acids such as palmitic acid, stearic acid, oleic acid, linoleic acid and octadecanoic acid may contribute to the anti-proliferative effect in keloid fibroblasts. CONCLUSIONS: The methanolic honey extraction has an antiproliferative effect on keloid fibroblasts and a range of volatile compounds has been identified from Tualang honey. The antiproliferative effect of keloid fibroblasts towards Tualang honey may involve cell signaling pathway. Identifying other volatile compounds from different organic solvents should be carried out in future.


Assuntos
Fatores Biológicos/farmacologia , Proliferação de Células/efeitos dos fármacos , Fibroblastos/citologia , Mel/análise , Queloide/tratamento farmacológico , Animais , Abelhas/metabolismo , Fatores Biológicos/análise , Fatores Biológicos/isolamento & purificação , Células Cultivadas , Fibroblastos/efeitos dos fármacos , Humanos , Queloide/fisiopatologia , Malásia , Metanol/química
17.
Int J Med Mushrooms ; 13(4): 327-33, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22164762

RESUMO

Forty isolates of Ophiocordyceps sinensis collected from Himalayan alpine meadows of Uttarakhand, India, and cultivated on Jhangora (Echinochloa crusgalli) grains were screened to identify the isolate(s) of high cordycepin content. The cultured mycelia were extracted with 50% methanol-chloroform and analyzed by HPTLC using chloroform:methanol (6:1 v/v) as mobile phase and densitometry scanning at 263 nm. Cordycepin varied from 0.002% to 0.029% was detected in twenty-one isolates. Compared to natural O. sinensis (0.004%, 0.006%), cordycepin was determined to be enhanced in twelve cultured samples.


Assuntos
Fatores Biológicos/análise , Desoxiadenosinas/análise , Alimento Funcional/análise , Hypocreales/química , Micélio/química , Fatores Biológicos/isolamento & purificação , Técnicas de Cultura , Desoxiadenosinas/isolamento & purificação , Hypocreales/crescimento & desenvolvimento , Índia , Micélio/crescimento & desenvolvimento
18.
Int J Med Mushrooms ; 13(4): 335-41, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22164763

RESUMO

The purpose of the study was to explore sixteen of the most popular edible species of wild-growing mushrooms as potential sources of antioxidants. Among the mushrooms tested, the highest total polyphenol contents, exceeding 100 mg/100 g fresh mass, were found in five mushrooms: Boletus chrysenteron, B. edulis, Leccinum scabrum, L. aurantiacum, and Macrolepiota procera. Antioxidant activity was measured with the FRAP, TEAC, DPPH scavenging ability and ferrous ions chelating ability assays. Results of the study show that wild mushrooms vary according to their antioxidant properties. The highest FRAP potentials, exceeding 1 mmol/100 g, were found in five species ofBoletales: Boletus edulis, B. chrysenteron, Leccinum scabrum, L. aurantiacum, and Suillus grevillei. TEAC values were from 1.07 to 4.01 mmol/100 g fresh mass. High TEAC values (>2.3 mmol/100 g) were found in Leccinum scabrum, L. aurantiacum, Macrolepiota procera, Boletus chrysenteron, and B. edulis. The DPPH radical scavenging effectiveness of mushroom extracts, expressed as EC50 values, was in range 2.91-13.86 mg/mL. Scavenging ability was the highest for B. edulis and B. chrysenteron. The metal chelating ability of mushroom extracts expressed as ECso values of chelating ability on ferrous ions were from 8.02 mg/mL in Cantharellus cibarius to 12.10 mg/mL in Suillus luteus. Among the mushrooms tested, Boletus chrysenteron and B. edulis were characterized by high scores of polyphenol contents and antioxidant activity in the FRAP, TEAC, and DPPH assays. These results place these culinary species of wild-growing mushrooms among products with considerable antioxidant potential.


Assuntos
Agaricales/química , Antioxidantes/análise , Fatores Biológicos/análise , Alimento Funcional/análise , Polifenóis/análise , Agaricales/classificação , Quelantes/análise
19.
Int J Med Mushrooms ; 13(4): 377-85, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-22164768

RESUMO

The effect of culinary-medicinal Royal Sun Agaricus (Agaricus brasiliensis) hot water extract on methyl methane sulfonate (MMS) induced mutagenicity/genotoxity in Drosophila melanogaster was studied using a quick and broadly applicable in vivo assay, i.e., the wing somatic mutation and recombination test. We used 2nd instar larvae, trans-heterozygous for the third chromosome recessive markers, i.e., multiple wing hairs (mvh) and flare-3 [flr (3)], and fed them for 24 h with the aqueous extract of A. brasiliensis. For antigenotoxicity studies a 24-h pretreatment with the extract was done, followed by a 48-h treatment of the then 3rd instar larvae with MMS. The frequency of mutations of the wing blade changes (i.e., of the number of wing spots of different sizes) induced in somatic cells was determined as a parameter of genetic changes of the wing imaginal discs. The results showed that A. brasiliensis extract did not cause any genotoxic or mutagenic effects. No antigenotoxic and/or protective effect against the induction of mutations by MMS was observed. Instead, a possible enhanced mitotic recombination frequency by MMS was seen after pretreatment of the larvae with A. brasiliensis extract. Possible mechanisms of action are discussed.


Assuntos
Agaricus/química , Antimutagênicos/farmacologia , Fatores Biológicos/farmacologia , Dano ao DNA/efeitos dos fármacos , Drosophila melanogaster/efeitos dos fármacos , Drosophila melanogaster/genética , Alimento Funcional/análise , Metanossulfonato de Metila/toxicidade , Mutagênicos/toxicidade , Animais , Antimutagênicos/análise , Antimutagênicos/isolamento & purificação , Fatores Biológicos/análise , Fatores Biológicos/isolamento & purificação , Drosophila melanogaster/crescimento & desenvolvimento , Testes de Mutagenicidade , Asas de Animais/efeitos dos fármacos , Asas de Animais/crescimento & desenvolvimento
20.
Wei Sheng Wu Xue Bao ; 51(6): 764-8, 2011 Jun.
Artigo em Zh | MEDLINE | ID: mdl-21866700

RESUMO

OBJECTIVE: Ganoderma lucidum was cultivated on non-medicinal parts of Salvia miltiorrhiza, Chrysanthemum morifolium, Ptatycodgn grandlfiorum, as all are Chinese traditional herbal medicines. We studied the changes of active ingredients and efficacies of the Ganoderma lucidum fruit bodies. METHODS: The agronomic characters, polysaccharide and terpene contents, acute toxicity and efficacy of Ganoderma lucidum grown on the non-medicinal part of the three materials were compared with that grown on the ordinary formula group (OF. G) which was composed of corn cob, cotton seed shell. RESULTS: Biological conversion efficiencies of the Ganoderma lucidum fruit body using non-medicinal parts were higher than that of using the ordinary formula group (OF. G), though growth periods became longer; Contents of active ingredients were all improved except that the terpene content of the Salvia miltiorrhiza group was decreased. Both polysaccharide and terpene from the Chrysanthemum morifolium group were the highest, contents of which were respectively 2.47% and 0.79%; Acute toxicity test showed that Ganoderma lucidum fruit bodies were all with low toxicities. Mice maximum tolerance dose were 100 g/kg weight. In hemolysin test and sleeping promotion test, the Chrysanthemum morifolium group showed better effect than the ordinary formula group (OF. G). In anti-fatigue test, only the ordinary formula group (OF. G) proved to be more effective. CONCLUSION: It's feasible to cultivate Ganoderma lucidum and active ingredients and efficacies of Ganoderma lucidum have been changed using the non-medicinal parts of Chinese medicinal herbs.


Assuntos
Fatores Biológicos/análise , Plantas Medicinais/química , Reishi/química , Reishi/crescimento & desenvolvimento , Animais , Fatores Biológicos/metabolismo , Fatores Biológicos/farmacologia , Chrysanthemum/química , Feminino , Masculino , Camundongos , Reishi/metabolismo , Salvia miltiorrhiza/química
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