Granulocyte-colony stimulating factor in experimental stroke and its effects on infarct size and functional outcome: a systematic review

Background Granulocyte-colony stimulating factor (G-CSF) shows promise as a treatment for stroke. This systematic review assesses G-CSF in experimental ischaemic stroke. Methods Relevant studies were identified with searches of Medline, Embase and PubMed. Data were extracted on stroke lesion size, neurological outcome and quality, and analysed using Cochrane Review Manager using random effects models; results are expressed as standardised mean difference (SMD) and odds ratio (OR). Results Data were included from 19 publications incorporating 666 animals. G-CSF reduced lesion size significantly in transient (SMD -1.63, p<0.00001) but not permanent (SMD -1.56, p=0.11) focal models of ischaemia. Lesion size was reduced at all doses and with treatment commenced within 4 hours of transient ischaemia. Neurological deficit (SMD -1.37, p=0.0004) and limb placement (SMD -1.88, p=0.003) improved with G-CSF; however, locomotor activity (>4 weeks post ischaemia) was not (SMD 0.76, p=0.35). Death (OR 0.27, p<0.0001) was reduced with G-CSF. Median study quality was 4 (range 0-7/8); Egger’s test suggested significant publication bias (p=0.001). Conclusions G-CSF significantly reduced lesion size in transient but not permanent models of ischaemic stroke. Motor impairment and death were also reduced. Further studies assessing dose-response, administration time, length of ischaemia and long-term functional recovery are needed.

Computational and experimental study of the behavior of cyano-based ionic liquids in aqueous solution

The solvation of cyano- (CN-) based ionic liquids (ILs) and their capacity to establish hydrogen bonds (H-bonds) with water was studied by means of experimental and computational approaches. Experimentally, water activity data were measured for aqueous solutions of ILs based on 1-butyl-3-methylimidazolium ([BMIM](+)) cation combined with one of the following anions: thiocyanate ([SCN](-)), dicyanamide ([DCA](-)), or tricyanomethanide ([TCM](-)), and of 1-ethyl-3-methylimidazolium tetracyanoborate ([EMIM][TCB]). From the latter data, water activity coefficients were estimated showing that [BMIM][SCN] and [BMIM][DCA], unlike [BMIM][TCM] and [EMIM][TCB], are able to establish favorable interactions with water. Computationally, the conductor like screening model for real solvents (COSMO-RS) was used to estimate the water activity coefficients which compare well with the experimental ones. From the COSMO-RS results, it is suggested that the polarity of each ion composing the ILs has a strong effect on the solvation phenomena. Furthermore, classical molecular dynamics (MD) simulations were performed for obtaining an atomic level picture of the local molecular neighborhood of the different species. From the experimental and computational data it is showed that increasing the number of CN groups in the ILs' anions does not enhance their ability to establish H-bonds with water but decreases their polarities, being [BMIM][DCA] and [BMIM][SCN] the ones presenting higher propensity to interact.

Measurements of activity coefficients at infinite dilution of organic solutes and water on polar imidazolium-based ionic liquids

The activity coefficients at infinite dilution, gamma(infinity)(13), of 55 organic solutes and water in three ionic liquids with the common cation 1-butyl-3-methylimidazolium and the polar anions Cl--,Cl- [CH3SO3](-) and [(CH3)(2)PO4](-), were determined by (gas + liquid) chromatography at four temperatures in the range (358.15 to 388.15) K for alcohols and water, and T = (398.15 to 428.15) K for the other organic solutes including alkanes, cycloalkanes, alkenes, cycloalkenes, alkynes, ketones, ethers, cyclic ethers, aromatic hydrocarbons, esters, butyraldehyde, acetonitrile, pyridine, 1-nitropropane and thiophene. From the experimental gamma(infinity)(13) values, the partial molar excess Gibbs free energy, (G) over bar (E infinity)(m), enthalpy (H) over bar (E infinity)(m), and entropy (S) over bar (E infinity)(m), at infinite dilution, were estimated in order to provide more information about the interactions between the solutes and the ILs. Moreover, densities were measured and (gas + liquid) partition coefficients (KL) calculated. Selectivities at infinite dilution for some separation problems such as octane/benzene, cyclohexane/benzene and cyclohexane/thiophene were calculated using the measured gamma(infinity)(13), and compared with literature values for N-methyl-2-pyrrolidinone (NMP), sulfolane, and other ionic liquids with a common cation or anion of the ILs here studied. From the obtained infinite dilution selectivities and capacities, it can be concluded that the ILs studied may replace conventional entrainers applied for the separation processes of aliphatic/aromatic hydrocarbons.

In vivo antileishmanial activity and chemical profile of polar extract from Selaginella sellowii

The polar hydroethanolic extract from Selaginella sellowii (SSPHE) has been previously proven active on intracellular amastigotes (in vitro test) and now was tested on hamsters infected with Leishmania (Leishmania) amazonensis (in vivo test). SSPHE suppressed a 100% of the parasite load in the infection site and draining lymph nodes at an intralesional dose of 50 mg/kg/day × 5, which was similar to the results observed in hamsters treated with N-methylglucamine antimonate (Sb) (28 mg/Kg/day × 5). When orally administered, SSPHE (50 mg/kg/day × 20) suppressed 99.2% of the parasite load in infected footpads, while Sb suppressed 98.5%. SSPHE also enhanced the release of nitric oxide through the intralesional route in comparison to Sb. The chemical fingerprint of SSPHE by high-performance liquid chromatography with diode-array detection and tandem mass spectrometry showed the presence of biflavonoids and high molecular weight phenylpropanoid glycosides. These compounds may have a synergistic action in vivo. Histopathological study revealed that the intralesional treatment with SSPHE induced an intense inflammatory infiltrate, composed mainly of mononuclear cells. The present findings reinforce the potential of this natural product as a source of future drug candidates for American cutaneous leishmaniasis.

Biological activity of Pyrrole-Imidazole polyamides in vivo

This thesis focuses on biological activity of pyrrole-imidazole polyamides in vivo. The work presented includes experiments underlining sequence selectivity of these compounds in living cells and potential methods to improve it. A large fraction of this thesis is devoted to activity of Py-Im in murine models of cancer. We investigated the pharmacokinetics and biodistribution of two compounds – targeted to 5'-WGGWCW-3' and 5'-WTWCGW-3' sequences – and characterized their activity by measuring their effects on tumor growth, gene expression in vivo and in tissue culture, and their effects on physiology of tumors. The initial theoretical studies suggested that a large fraction of genomic sites are bound by Py-Im polyamides non-specifically and experimental data shows that the programmed binding sequence is not a sole determinant of the patterns of gene regulation. Despite the likely presence of non-specific effects of Py-Im polyamides in living cells, in vivo administration of Py-Im polyamides resulted in tolerable host toxicity and anti-tumor activity. Py-Im polyamide targeted to Estrogen Receptor Response Element showed downregulation of ER-driven gene expression in tumor cells, while the compound targeted to hypoxia response element reduced vascularization of tumors and their growth rate, induced apoptosis of cells in hypoxic areas and reduced expression of proangiogenic and prometastatic factors. Further studies, showed that polyamides distributed to many of the tested tissues and their FITC-conjugates showed nuclear uptake. The gene expression effects were also present in murine tissues, such as liver and kidneys, indicating a potential for use for Py-Im polyamides in non-cancerous diseases.

Natural recovery of the benthic nematode assemblages associated to Zostera noltii seagrass beds after physical disturbance caused by digging harvest activity

Seagrass beds are productive ecosystems that maintain high levels of biodiversity, making them susceptible to anthropogenic pressures such as bivalve harvesting. Nematodes are considered great ecological indicators as changes in their density, diversity and structure may represent changes in the environment. This experimental fieldwork aimed to assess the impact of the bivalve harvesting on the nematodes assemblage of a seagrass bed in the Mira estuary by simulating the digging activity. Two plots were subjected to the digging (D1 and D19) and two plots were control (C11 and C18). The sampling took place in five occasions: T0 – before digging; T1 – 14 days; T2 – 45 days; T3 – 75 days; and T4 – 165 days after digging. The results showed no significant difference in the nematode assemblages’ density, diversity and trophic composition between treatments and sampling times, evidencing their high tolerance for naturally stressed environments and to the level of digging they were exposed; Recuperação natural das comunidades de nematodes bentónicos associados aos povoamentos de Zostera noltii após atividade de marisqueio Resumo: As pradarias marinhas são ecossistemas produtivos que suportam elevados níveis de biodiversidade, pelo que estão sujeitos a pressões antropogénicas. Os nematodes são bons indicadores ecológicos pois respondem rapidamente a qualquer perturbação por alterações na densidade, diversidade e estrutura. Este trabalho experimental teve como finalidade o estudo da recuperação natural das comunidades de nematodes associados aos povoamentos de Zostera noltii pela simulação da atividade de marisqueio. Dois plots foram sujeitos a revolvimento (D1 e D19) e dois plots serviram como controlo (C11 e C18) e foram efetuadas amostragens em cinco ocasiões: T0 – antes do revolvimento; T1 – 14 dias; T2 – 45 dias; T3 – 75 dias; e T4 – 165 dias após revolvimento. Os resultados obtidos não mostraram diferenças significativas na diversidade, densidade e composição trófica das comunidades de nematodes entre tratamentos e tempos de amostragem, evidenciando a sua elevada tolerância a ambientes naturalmente dinâmicos e ao nível de revolvimento a que foram expostas.

The effect of chaperone compounds on glucocerebrosidase stability and activity in fibroblasts from Gaucher Disease patients with different genotypes

Dissertação de mestrado, Ciências Biomédicas, Departamento de Ciências Biomédicas e Medicina, Universidade do Algarve, 2014

Patterns of sedentary behavior : insights from observational and experimental studies on body composition and energy expenditure

It is recognized that sedentary behavior (SB) has deleterious effects on numerous health outcomes and it appears that physiological mechanisms underlying these harms are distinct from the ones explaining moderate-to-vigorous physical activity (MVPA) benefits. Sedentary behavior represents a large portion of human’s life and is increasing with technological development. A new current of opinion supports the idea that the manner SB is accumulated plays an important role. This dissertation presents six research studies conducted under the scope of SB. In the methodological area, the first study highlighted the magnitude of potential errors in estimating SB and its patterns from common alternative methods (accelerometer and heart rate monitor) compared to ActivPAL. This study presented the accelerometer as a valid method at a group level. Two studies (2 and 5) were performed in older adults (the most sedentary group in the population) to test the associations for SB patterns with abdominal obesity using accelerometry. The findings showed positive graded associations for prolonged sedentary bouts with abdominal obesity and showed that those who interrupted SB more frequently were less likely to present abdominal obesity. Therefore, public health recommendations regarding breaking up SB more often are expected to be relevant. The associations between sedentary patterns and abdominal obesity were independent of MVPA in older adults. However, the low MVPA in this group makes it unclear whether this independent relationship still exists if highly active persons are analysed. Study 3 inovates by examining the association of SB with body fatness in highly trained athletes and found SB to predict total fat mass and trunk fat mass, independently of age and weekly training time. Study 4 also brings novelty to this research field by quantifying the metabolic and energetic cost of the transition from sitting to standing and then sitting back down (a break), informing about the modest energetic costs (0.32 kcal·min−1). Finally, from a successful multicomponent pilot intervention to reduce and break up SB (study 6), an important behavioral resistance to make more sit/stand transitions despite successfully reducing sitting time (~ 1.85 hours·day-1) was found, which may be relevant to inform future behavioral modification programs. The present work provides observational and experimental evidence on the relation for SB patterns with body composition outcomes and energy regulation that may be relevant for public health interventions.

Antilipolytic and hypotriglyceridemic effects of dietary Salvia triloba Lf (Lamiaceae) in experimental rats

Purpose: Pancreatic triacylglycerol lipase (PL) is a noteworthy pharmacological target for the management of dyslipidemia, and diabetes and obesity. This study was aimed to evaluate the modulatory effects of Salvia triloba L.f. (Lamiaceae) leaves methanol extract (ME) on a high fat diet (HFD)-induced hypertriglyceridemia in rats, with complementary in vitro evaluation of sage PL-inhibitory potential. Methods: Pre-induction of HFD hypertriglyceridemia sage leaves ME (750 mg/kg) was orally supplemented (via gastric intubation) to overnight fasting rats (n = 5). Potential plant modulation of PL was also quantified in vitro by a colorimetric assay (n = 3). For comparison, the effect of Orlistat was similarly evaluated as reference standard. Results: Compared to Orlistat, supplementation of S. triloba at a dose of 750 mg/kg b.wt significantly reversed the HFD-induced postprandial hypertriglyceridemia in experimental overnight fasting rats (p < 0.001 vs. HFD rats). Dietary sage caused 66.4 % reduction in plasma triglycerides. Compared to Orlistat which exerted antilipolytic activity, with half-maximal inhibitory concentration (IC50) of 0.114 ± 0.004 μg/mL), sage inhibited PL activity in vitro in a dose-dependent manner IC50 of 100.80 ± 9.07 μg/mL) Conclusion: Sage has dual hypotriglyceridemic and antilipolytic properties which indicate that it can potentially be used to suppress body weight gain.

Spatial and Temporal Patterns in the Distribution, Behavior, and Activity of Fishes in Canals of the Everglades

Landscape characteristics, disturbances, and temporal variability influence predator-prey relationships, but are often overlooked in experimental studies. In the Everglades, seasonal disturbances force the spatial overlap of predators and prey, potentially increasing predation risk for prey. This study examined seasonal and diel patterns of fish use of canals and assessed predation risk for small fishes using an encounter rate model. I deployed an imaging sonar in Everglades canals to quantify density and swimming speeds of fishes, and detect anti-predator behaviors by small fishes. Generally, seasonal declines of marsh water-levels increased the density of large fishes in canals. Densities of small and large fishes were positively correlated and, as small-fish density increased, schooling frequency also increased. At night, schools disbanded and small fishes were observed congregating along the canal edge. The encounter rate model predicted highest predator-prey encounters during the day, but access to cover may reduce predation risk for small fishes.

Deficit of quantal release of GABA in experimental models of temporal lobe epilepsy

Because GABA (gamma-aminobutyric acid) receptor-mediated inhibition controls the excitability of principal neurons in the brain, deficits in GABAergic inhibition have long been favored to explain seizures. In an experimental model of temporal lobe epilepsy, we have identified a deficit of inhibition in presynaptic GABAergic terminals characterized by decreased GABA quantal activity associated with reduced synaptic vesicle density. This decrease in vesicle number primarily seems to affect the reserve pool, rather than the docked or the readily releasable pool.

Dynamical modeling of biogas production supported by an experimental anaerobic digestion process

This paper presents the development of a combined experimental and numerical approach to study the anaerobic digestion of both the wastes produced in a biorefinery using yeast for biodiesel production and the wastes generated in the preceding microbial biomass production. The experimental results show that it is possible to valorise through anaerobic digestion all the tested residues. In the implementation of the numerical model for anaerobic digestion, a procedure for the identification of its parameters needs to be developed. A hybrid search Genetic Algorithm was used, followed by a direct search method. In order to test the procedure for estimation of parameters, first noise-free data was considered and a critical analysis of the results obtain so far was undertaken. As a demonstration of its application, the procedure was applied to experimental data.

Enzymatic activity on sediments and nematode assemblage responses during seagrass beds habitat recovery following the disturbance of the traditional digging activity of bivalves harvesting.

Sediment digging is an anthropogenic activity connected to the exploitation of living resources in estuarine and marine environments. The knowledge on the functional responses of the benthic assemblages to the physical disturbance is an important baseline to understand the ecological processes of the habitat recovery and restoration and to develop tools for the management of the harvesting activities. To investigate the effects of the digging activity of the bivalves on Zostera noltii seagrass beds a manipulative field experiment was conducted that included the enzymatic activity of sediments and the associated nematode assemblages. Four plots (two undisturbed serving as control and two dug to collect bivalves - treatment) with 18 subplots were randomly located at seagrass beds in the Mira estuary at the SW coast of Portugal. Samples were randomly and unrepeatably collected from three subplots of each plot in five different occasions, before sediment digging (T0) up to six months after disturbance (T5). Microbial activity in sediments was assess by determining the extracelular enzymatic activity of six hydrolytic enzymes (sulfatase, phosphatase, b -N-acetilglucosaminidase, b-glucosidase, urease, protease) and two oxidoreductases (phenol oxidase and peroxidase). The microbial community status was also assessed through the measurement of dehydrogenase, which reflects microbial respiration. The nematode assemblages composition, biodiversity and trophic composition at different sampling occasions were also analyzed. The fluorometric and biochemical parameters analysed of the Z. noltii plants during the experimental period showed a recovery of the seagrass beds, and it was detected an increase of the enzymatic activity of the sediments after disturbance. The nematodes assemblages were similar in all sampling occasions. The seagrass beds and the nematodes assemblages associated showed a high resilience to the stress caused by the traditional bivalves digging activity. The obtained results allow the development of a management programme for the commercial fishing activity to maintain the good environmental status and minimized the secondary environmental effects on marine and estuarine habitats through the establishment of a baseline for the regulation of the harvesting frequency.

Influence of nitrogen sources on the enzymatic activity and grown by Lentinula edodes in biomass Eucalyptus benthamii.

Lignocelulose é o componente mais abundante do meio ambiente e recurso orgânico renovável no solo. Alguns fungos filamentosos têm desenvolvido a habilidade de degradar e utilizar celulose, hemicelulose e lignina como fonte de energia. O objetivo deste trabalho foi analisar o efeito de três fontes de nitrogênio (sulfato de amônio, nitrato de potássio e farelo de soja) na atividade enzimática de Lentinula edodes EF 50 utilizando como substrato serragem de E. benthamii. Foi aplicado um planejamento experimental de mistura com três repetições no ponto central constituído de sete tratamentos (T) de iguais concentrações em nitrogênio de sulfato de amônia, nitrato de potássio e farinha de soja cozida. Foram determinadas a atividade enzimática da avicelase, carboximetilcelulase, β-glicosidase, xilanases e manganês peroxidase. Foram avaliados o teor de umidade, pH, atividade de água (aw) e análise qualitativa do crescimento micelial em 8 tempos de cultivo. Os resultados mostraram efeito negativo na produção das enzimas nos tratamentos com máxima concentração de sulfato de amônia e nitrato de potássio. Os tratamentos com farinha de soja cozida expressaram maiores atividades enzimáticas, nos tempos de 3, 6 e 9 dias de cultivo exceto na atividade do manganês peroxidase. A maior produção foi observada no tratamento com sulfato de amônia e farinha de soja cozida (83.86 UI.L?1) em 20 dias de cultivo.