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2.
Int J Mass Spectrom ; 301(1-3): 7-11, 2011 Mar 30.
Artigo em Inglês | MEDLINE | ID: mdl-21499518

RESUMO

Microwave assisted acid cleavage was applied directly to intact adenovirus type 5 to achieve denaturation and proteolysis in a single reaction. The speed of the digestion, coupled with the simplicity of MALDI analysis, allowed peptide products to be profiled in less than 5 min. Identification of peptides from a range of proteins by MALDI-TOF confirms that both denaturation and proteolysis were achieved using low concentrations of acetic acid (12.5%) and short incubations (1.5 to 2 min) at high temperatures (140° C). These conditions favor production of peptides that carry Asp on their C-termini. When this cleavage reaction is carried out in highly enriched H(2) (18)O, a single atom of (18)O is introduced site-specifically into the carboxyl terminal. The labeling reaction is applied to label reporter peptides from human adenovirus type 5 harvested from HeLa cells. Small peptide products of endogenous processing were also observed.

3.
Psychoneuroendocrinology ; 124: 105064, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-33260082

RESUMO

Previous research suggests that maternal prenatal psychological distress (PPD) is related to altered cortisol reactivity in the exposed child. There are indications for the sex differences in vulnerability for prenatal adversities that depend on the exposure and child outcome. Still, it is not known whether the association between maternal PPD and infant cortisol stress response is moderated by sex. In addition, the recovery phase of the cortisol stress response has not been given as much attention as reactivity. Our aim was to study the sex differences in the associations between self-reported maternal prenatal depressive-, anxiety- and pregnancy-related anxiety symptoms through gestational weeks 14, 24 and 34 and the saliva cortisol reactivity to and recovery from the acute stress among 10-week-old infants. The study population comprised of 363 mother-infant pairs from the FinnBrain Birth Cohort Study. We found evidence for sex-dependent associations between PPD exposure and infant cortisol response. A less steep recovery slope (-10 % per one SD increase in PPD [95 % CI = -18 to -2 %] and -8 % [-16 to 0 %] depending on the exposure) and a possibly less steep reactivity slope (-14 % [95 % CI = -25 to 0 %] and -10 % [-21 to 3 %]) were associated with higher PPD exposure in females. Of the PPD measures, the strongly intercorrelated, and thus combined, depressive and anxiety symptom score provided the most robust prediction of infant cortisol recovery. Our results demonstrate sexually dimorphic alterations in the functioning of the infant hypothalamus-pituitary-adrenal axis and especially in the functioning of the negative feedback loop of the axis after prenatal PPD exposure among healthy babies.


Assuntos
Hidrocortisona , Efeitos Tardios da Exposição Pré-Natal , Criança , Estudos de Coortes , Feminino , Humanos , Lactente , Masculino , Gravidez , Saliva , Caracteres Sexuais , Estresse Psicológico
5.
PLoS One ; 13(9): e0204207, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30222764

RESUMO

Fatty acid (FA) signatures (FAS) are important tools to assess the foraging ecology of wild animals. The present study was conducted to assess how well the general FAS and the proportions of individual FA are preserved in fat samples stored at different temperatures (-196, -80, -20, +4 and +20°C). Using three species (laboratory rat, American mink and rainbow trout), FAS were determined immediately upon sampling. Thereafter, eight subsamples per storage temperature from the inner part of the sample unaffected by oxygen and light were re-analyzed after 1, 2, 3, 7, 28, 84 and 168 days. Each time the remaining sample was sealed in its vial after replacing air with nitrogen gas. The results were tested with the mixed model and discriminant analyses. Generally, the FAS were well preserved regardless of storage temperature, and only a few major FA showed significant changes even after the 6-month period at room temperature. After an initial first-day change in proportions, presumably due to post-mortem enzymatic activities, the remaining minor changes could not be clearly attributed to either further autolysis, decomposition or autoxidation. In the discriminant analysis, the species-specific differences dominated and remained distinct even after 6 months. Furthermore, the analysis mostly classified the samples preserved at sub- and above-freezing temperatures separate from each other, and the general deviation from the initial analysis results was present as early as after 1 day. If FAS are to be analyzed in a very precise manner, the analysis should be performed immediately upon sampling. However, FAS remain adequately reliable for long periods of time even without preservation in deep freeze, widening the availability of potential samples for studies on foraging ecology and related disciplines.


Assuntos
Cromatografia Gasosa/métodos , Ácidos Graxos/análise , Manejo de Espécimes/métodos , Animais , Análise Discriminante , Feminino , Masculino , Vison , Oncorhynchus mykiss , Ratos , Especificidade da Espécie , Temperatura , Fatores de Tempo
6.
Mar Pollut Bull ; 52(2): 149-61, 2006 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-16212986

RESUMO

The concentrations and composition profiles of polychlorinated dibenzo-p-dioxins and dibenzofurans, polychlorinated biphenyls, polybrominated diphenyl ethers and polychlorinated naphthalenes were determined in herring individuals collected from the commercial catches of the Bothnian Sea, northern Baltic. The median age of herring was 5.0 years and in the muscle the median toxic equivalent concentration of PCDD/Fs was 5.6 WHO(PCDD/F)-TEQ pg/g fresh weight (fw) and that of PCBs 2.7 WHO(PCB)-TEQ pg/g fw. The median sum concentration of PBDEs was 1.4 ng/g fw and that of PCNs 0.1 ng/g fw. Differences in age-dependent accumulation between the organohalogen groups and individual congeners were major. In the Bothnian Sea the content of organohalogen compounds in herring is obviously elevated due to the availability and large proportion of Mysis crustaceans in their diet. More intensive fishing could reduce the concentrations of organohalogens, including the abundant, accumulative and toxic 2,3,4,7,8-PeCDF congener in the Baltic herring catch.


Assuntos
Monitoramento Ambiental , Peixes/fisiologia , Hidrocarbonetos Halogenados/análise , Poluentes Químicos da Água/análise , Fatores Etários , Animais , Benzofuranos/análise , Tamanho Corporal , Feminino , Peixes/crescimento & desenvolvimento , Masculino , Naftalenos/análise , Oceanos e Mares , Éteres Fenílicos/análise , Bifenilos Policlorados/análise , Dibenzodioxinas Policloradas/análogos & derivados , Dibenzodioxinas Policloradas/análise
8.
Sci Total Environ ; 340(1-3): 1-11, 2005 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-15752488

RESUMO

Sunlight or ultraviolet light irradiation of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) in the presence of vegetable oil offers a potential method for the cleanup of contaminated soil. In this study, the effects of different types of vegetable oils on the photochemical degradation of 1,2,3,4,6,7,8-heptachlorodibenzofuran and heptachlorodibenzo-p-dioxin (1,2,3,4,6,7,8-HpCDF/HpCDD) were investigated in the laboratory. Using a blacklight lamp as a source of ultraviolet light, 93-100% of 1,2,3,4,6,7,8-HpCDF degraded in 60 min in rapeseed oil, extra virgin olive oil and olive oil. Less degradation occurred in palm oil (59%), toluene (39%) and hexane (20%). The better degradation in vegetable oils in comparison with organic solvents was attributed to the photooxidation of lipids producing hydrogen for PCDD/F dechlorination. In addition to the hydrogen donor capacity, permeability of ultraviolet light was involved in the differences between vegetable oils. alpha-Tocopherol and chlorophyll did not influence the performance of oil at concentrations normally present in vegetable oils, whereas beta-carotene had an inhibitory effect on the degradation of 1,2,3,4,6,7,8-HpCDF. Up to 28% of the degradation products of 1,2,3,4,6,7,8-HpCDF were formed via the dechlorination pathway. Products included both toxic (2,3,7,8-chlorinated) and non-toxic PCDD/Fs, the toxic PCDD/Fs being more stable. Irradiation of 1,2,3,4,6,7,8-HpCDD yielded only non-toxic dechlorination products. Polychlorinated hydroxybiphenyls (OH-PCBs), polychlorinated dihydroxybiphenyls (DOH-PCBs) and polychlorinated hydroxydiphenylethers (OH-PCDEs) containing one to seven chlorine atoms were not detected in irradiated HpCDF/HpCDD samples.


Assuntos
Benzofuranos/química , Óleos de Plantas/química , Dibenzodioxinas Policloradas/análogos & derivados , Dibenzodioxinas Policloradas/química , Poluentes do Solo/análise , Dibenzofuranos Policlorados , Poluição Ambiental/prevenção & controle , Fotólise , Controle de Qualidade , Solubilidade , Raios Ultravioleta
9.
Water Res ; 38(17): 3769-79, 2004 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-15350429

RESUMO

We studied the changes in water quality and formation of biofilms occurring in a pilot-scale water distribution system with two generally used pipe materials: copper and plastic (polyethylene, PE). The formation of biofilms with time was analysed as the number of total bacteria, heterotrophic plate counts and the concentration of ATP in biofilms. At the end of the experiment (after 308 days), microbial community structure, viable biomass and gram-negative bacterial biomass were analysed via lipid biomarkers (phospholipid fatty acids and lipopolysaccharide 3-hydroxy fatty acids), and the numbers of virus-like particles and total bacteria were enumerated by SYBR Green I staining. The formation of biofilm was slower in copper pipes than in the PE pipes, but after 200 days there was no difference in microbial numbers between the pipe materials. Copper ion led to lower microbial numbers in water during the first 200 days, but thereafter there were no differences between the two pipe materials. The number of virus-like particles was lower in biofilms and in outlet water from the copper pipes than PE pipes. Pipe material influenced also the microbial and gram-negative bacterial community structure in biofilms and water.


Assuntos
Biofilmes/crescimento & desenvolvimento , Cobre , Plásticos , Microbiologia da Água , Abastecimento de Água , Engenharia , Monitoramento Ambiental , Arquitetura de Instituições de Saúde , Controle de Qualidade , Água/química
10.
Eur J Mass Spectrom (Chichester) ; 9(3): 195-201, 2003.
Artigo em Inglês | MEDLINE | ID: mdl-12939497

RESUMO

In order to investigate the systematic changes in fragmentation behavior of poly(methyl methacrylate) (PMMA) with increasing molecular weight, alkali-metal cationized PMMA 20-mer, 60-mer and 100-mer were selected for post-source decay (PSD) fragmentation study by matrix-assisted laser desorption/ionization time-of-flight (MALDI-ToF) mass spectrometry. PMMA polymers were cationized with lithium, potassium and cesium cations to explore the influence of the cation size on the fragmentation behavior of the polymers. All PMMA polymers could be fragmented by MALDI-PSD and fragmentation of the MALDI ionized synthetic polymer of molecular weight 10 kDa is reported here for the first time. It was shown that an increasing molecular weight of the PMMA chain required an increase in the size of the cation to improve the intensity and the number of the fragments in the PSD spectrum. Some instrumental parameters had to be optimized prior to a successful PSD analysis of the largest PMMA polymers.

11.
PLoS One ; 9(12): e114490, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25473947

RESUMO

In vitro toxicological studies together with atomistic molecular dynamics simulations show that occupational co-exposure with C60 fullerene may strengthen the health effects of organic industrial chemicals. The chemicals studied are acetophenone, benzaldehyde, benzyl alcohol, m-cresol, and toluene which can be used with fullerene as reagents or solvents in industrial processes. Potential co-exposure scenarios include a fullerene dust and organic chemical vapor, or a fullerene solution aerosolized in workplace air. Unfiltered and filtered mixtures of C60 and organic chemicals represent different co-exposure scenarios in in vitro studies where acute cytotoxicity and immunotoxicity of C60 and organic chemicals are tested together and alone by using human THP-1-derived macrophages. Statistically significant co-effects are observed for an unfiltered mixture of benzaldehyde and C60 that is more cytotoxic than benzaldehyde alone, and for a filtered mixture of m-cresol and C60 that is slightly less cytotoxic than m-cresol. Hydrophobicity of chemicals correlates with co-effects when secretion of pro-inflammatory cytokines IL-1ß and TNF-α is considered. Complementary atomistic molecular dynamics simulations reveal that C60 co-aggregates with all chemicals in aqueous environment. Stable aggregates have a fullerene-rich core and a chemical-rich surface layer, and while essentially all C60 molecules aggregate together, a portion of organic molecules remains in water.


Assuntos
Poluentes Ocupacionais do Ar/toxicidade , Fulerenos/toxicidade , Acetofenonas/química , Acetofenonas/toxicidade , Poluentes Ocupacionais do Ar/química , Benzaldeídos/química , Benzaldeídos/toxicidade , Álcool Benzílico/química , Álcool Benzílico/toxicidade , Linhagem Celular Tumoral , Cresóis/química , Cresóis/toxicidade , Interações Medicamentosas , Fulerenos/química , Humanos , Interações Hidrofóbicas e Hidrofílicas , Interleucina-1beta/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/fisiologia , Simulação de Dinâmica Molecular , Termodinâmica , Tolueno/química , Tolueno/toxicidade , Fator de Necrose Tumoral alfa/metabolismo
12.
Anal Chim Acta ; 699(1): 73-80, 2011 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-21704760

RESUMO

We have studied the matrix effect within direct analysis of benzodiazepines and opioids from urine with desorption electrospray ionization-mass spectrometry (DESI-MS) and desorption atmospheric pressure photoionization-mass spectrometry (DAPPI-MS). The urine matrix was found to affect the ionization mechanism of the opioids in DAPPI-MS favoring proton transfer over charge exchange reaction. The sensitivity for the drugs in solvent matrix was at the same level with DESI-MS and DAPPI-MS (LODs 0.05-6 µg mL(-1)) but the decrease in sensitivity due to the urine matrix was higher with DESI (typically 20-160-fold) than with DAPPI (typically 2-15-fold) indicating better matrix tolerance of DAPPI over DESI. Also in MS/MS mode, DAPPI provided better sensitivity than DESI for the drugs in urine. The feasibility of DAPPI-MS/MS was then studied in screening the same drugs from five authentic, forensic post mortem urine samples. A reference measurement with gas chromatography-mass spectrometry (GC-MS) (including pretreatment) revealed 16 findings from the samples, whereas with DAPPI-MS/MS after sample pretreatment, 15 findings were made. Sample pretreatment was found necessary, since only eight findings were made from the same samples untreated.


Assuntos
Analgésicos Opioides/urina , Benzodiazepinas/urina , Espectrometria de Massas por Ionização por Electrospray/métodos , Detecção do Abuso de Substâncias/métodos , Pressão Atmosférica , Medicina Legal , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Solventes/química , Espectrometria de Massas por Ionização por Electrospray/instrumentação
13.
J Mol Biol ; 381(1): 89-101, 2008 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-18586268

RESUMO

The biotin repressor is an allosterically regulated, site-specific DNA-binding protein. Binding of the small ligand bio-5'-AMP activates repressor dimerization, which is a prerequisite to DNA binding. Multiple disorder-to-order transitions, some of which are known to be important for the functional allosteric response, occur in the vicinity of the ligand-binding site concomitant with effector binding to the repressor monomer. In this work, the extent to which these local changes are coupled to additional changes in the structure/dynamics of the repressor was investigated using hydrogen/deuterium exchange coupled to mass spectrometry. Measurements were performed on the apo-protein and on complexes of the protein bound to four different effectors that elicit a range of thermodynamic responses in the repressor. Global exchange measurements indicate that binding of any effector to the intact protein is accompanied by protection from exchange. Mass spectrometric analysis of pepsin-cleavage products generated from the exchanged complexes reveals that the protection is distributed throughout the protein. Furthermore, the magnitude of the level of protection in each peptide from hydrogen/deuterium exchange correlates with the magnitude of the functional allosteric response elicited by a ligand. These results indicate that local structural changes in the binding site that occur concomitant with effector binding nucleate global dampening of dynamics. Moreover, the magnitude of dampening of repressor dynamics tracks with the magnitude of the functional response to effector binding.


Assuntos
Dobramento de Proteína , Proteínas Repressoras/química , Proteínas Repressoras/metabolismo , Transdução de Sinais , Monofosfato de Adenosina/metabolismo , Regulação Alostérica , Sequência de Aminoácidos , Medição da Troca de Deutério , Ligantes , Modelos Moleculares , Dados de Sequência Molecular , Ligação Proteica , Estrutura Quaternária de Proteína , Estrutura Terciária de Proteína , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
14.
Mol Microbiol ; 66(3): 787-800, 2007 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-17908204

RESUMO

The ESX-1 secretion system plays a critical role in the virulence of Mycobacterium tuberculosis and M. marinum. To date, three proteins are known to be secreted by ESX-1 and necessary for virulence, two of which are CFP-10 and ESAT-6. The ESX-1 secretion and the virulence mechanisms are not well understood. In this study, we have examined the M. marinum secretomes and identified four proteins specific to ESX-1. Two of those are CFP-10 and ESAT-6, and the other two are novel: MM1553 (homologous to Rv3483c) and Mh3881c (homologous to Rv3881c). We have shown that Mh3881c, CFP-10 and ESAT-6 are co-dependent for secretion. Mh3881c is being cleaved at close to the C-terminus during secretion, and the C-terminal portion is critical to the co-dependent secretion, the ESAT-6 cellular levels, and interaction with ESAT-6. The co-dependent secretion is required for M. marinum intracellular growth in macrophages, where the Mh3881c C-terminal portion plays a critical role. The role of the co-dependent secretion in intracellular growth correlates with its role in inhibiting phagosome maturation. Both the secretion and the virulence defects of the Mh3881c mutant are complemented by Mh3881c or its M. tuberculosis homologue Rv3881c, suggesting that in M. tuberculosis, Rv3881c has similar functions.


Assuntos
Antígenos de Bactérias/metabolismo , Proteínas de Bactérias/metabolismo , Mycobacterium tuberculosis/metabolismo , Fagossomos/metabolismo , Animais , Western Blotting , Eletroforese em Gel Bidimensional , Macrófagos/citologia , Macrófagos/metabolismo , Macrófagos/microbiologia , Espectrometria de Massas , Camundongos , Mycobacterium tuberculosis/patogenicidade , Virulência
15.
Anal Chem ; 74(16): 4250-8, 2002 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-12199599

RESUMO

Polyesters prepared from the same diol, 2-butyl-2-ethyl-1,3-propanediol, but different phthalic acid isomers, phthalic, isophthalic, and terephthalic acid, were characterized by collision-induced dissociation electrospray ionization Fourier transform ion cyclotron resonance (CID-ESI-FT-ICR) and postsource-decay matrix-assisted laser desorption/ionization time-of-flight (PSD-MALDI-TOF) mass spectrometry. Sodiated dihydroxyl-terminated polyester oligomers containing five repeating units at m/z 1634 were selected as precursor ions for dissociation studies. Two main mechanisms occurred in the fragmentation of all of the polyesters, since dissociation of the oligomers was initiated by hydrogen rearrangement or transesterification reactions. Polyesters prepared from different phthalic acid isomers could be distinguished by their fragmentation behavior. Polyester prepared from phthalic acid was easily identified by using both CID-ESI-FT-ICR and PSD-MALDI-TOF mass spectrometry. However, distinguishing between the polyesters prepared from isophthalic and terephthalic acid succeeded marginally only with CID-ESI-FT-ICR mass spectrometry. Molecular dynamics calculations were used to obtain an idea of the fragmentation behavior of the polyesters. The low-energy structures of the precursor ions were determined, and the coordination of the oxygen atoms of the polyester oligomers to the sodium cation was examined more closely. Both the experimental and the theoretical studies showed that the sodium ion affinity of polyester changed with the phthalic acid isomer.

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