Imidazolium-based ionic liquids binding to DNA: Mechanical effects and thermodynamics of the interactions.

We performed a robust characterization of the molecular interactions between the DNA molecule and two imidazolium-based ionic liquids (ILs): 1-Butyl-3-methylimidazolium chloride ([bmim]Cl) and 1-Octyl-3-methylimidazolium chloride ([omim]Cl), using single molecule approaches (optical and magnetic tweezers) and bulk techniques (isothermal titration calorimetry and conductivity measurements). Optical and magnetic tweezers allowed us to obtain the changes on the mechanical properties of the DNA complexes formed with both ILs, as well as the relevant physicochemical (binding) parameters of the interaction. Despite the weak binding measured between DNA and the two ILs, we identify a transition on the regime of polymer elasticity of the complexes formed, which results in a relevant DNA compaction for high IL concentrations. In addition, isothermal titration calorimetry and conductivity complemented the single molecule investigation, giving a complete thermodynamic characterization of the interactions and allowing the identification of the most relevant driving forces at various different concentration ranges of the ILs. Based on the results obtained with all the employed techniques, we propose a model for the binding schemes involving DNA and both [bmim]Cl and [omim]Cl.

Inflammatory profile of apical periodontitis associated with liver fibrosis in rats: histological and immunohistochemical analysis.

AIM: To investigate the effects of liver fibrosis (LF) on the pro-inflammatory mediators and periapical bone resorption of apical periodontitis (AP) in rats. METHODOLOGY: Forty male Wistar rats were distributed into four groups: C - control, AP - rats with AP, LF - rats with LF, AP + LF - rats with AP and LF. LF was induced by carbon tetrachloride administration for 8 weeks and surgical bile duct ligation for 4 weeks; AP was induced in the teeth of rats by dental pulp exposure to the oral environment for 30 days. Jaws and livers were removed after euthanasia. Haematoxylin and Eosin (H&E) and Picrosirius Red (PSR) staining were used to confirm fibrosis in the livers. The jaws were analysed using H&E staining, immunohistochemical assays of interleukin (IL)-1ß, IL-6 and tumour necrosis factor-alpha (TNF-α). Student's t-test and Mann-Whitney's U-test were used for statistical analysis (P < 0.05). RESULTS: Inflammatory infiltrate was moderate in the AP group and severe in the AP + LF group (P < 0.05). Periapical bone resorption was significantly larger in the AP + LF group compared with the AP group (P < 0.05). IL-1ß, IL-6 and TNF-α levels were significantly higher in AP + LF group when compared to the AP group (P < 0.05). CONCLUSION: More intense inflammatory infiltrate, greater amounts of pro-inflammatory cytokines and increased periapical bone resorption were observed in the presence of liver fibrosis in rats with exposed pulps.

Relationship between apical periodontitis and atherosclerosis in rats: lipid profile and histological study.

AIM: To investigate the relationship between apical periodontitis and atherosclerosis in rats by lipid profile and carotid artery intima tunic measurement, and histological and histometric evaluation of periapical lesions. METHODOLOGY: Forty male Wistar rats were allocated into four groups: control (C), with apical periodontitis (AP), with atherosclerosis (AT) and with AP and AT (AP + AT). Atherosclerosis was induced using a high-lipid diet associated with a surgical ligature in the carotid artery and a super dosage of vitamin D3 . AP was induced via pulp exposure to the oral environment. At 45 and 75 days, serum levels of total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) were measured. The maxillary and mandibular jaws and carotid artery were collected and processed for histological analysis. The Kruskal-Wallis or Mann-Whitney test was performed for nonparametric data, and the Tukey's or Student's t-test was performed for parametric data (P < 0.05). RESULTS: In nonatherosclerotic animals, the induction of apical periodontitis increased TG levels significantly, from 63.1 ± 11.4 mg dL-1 in group C to 88.2 ± 7.9 mg dL-1 in the AP group (P < 0.05). The induction of AP was associated with a trend for higher TC and LDL-C levels in atherosclerotic animals (P > 0.05); however, it only significantly increased TG levels, from 93.2 ± 18.0 mg dL-1 in AT group to 121.9 ± 14.5 mg dL-1 in the AP + AT group (P < 0.05). Animals in the AP + AT group had a 36.5% increase in the thickness of the carotid intima tunic when compared with the AT group (P < 0.05). The intensity of the inflammatory infiltrate was significantly larger in the AP + AT group when compared with AP group (P < 0.05). The AP + AT group exhibited significantly greater alveolar bone loss, with a periapical lesion size of 206.4 ± 56.3 × 104 µm2 , compared with 151.4 ± 49.1 × 104 µm2 in the AP group (P < 0.05). CONCLUSION: Apical periodontitis influenced triglyceride levels, increasing them even in the absence of atherosclerosis, and influenced the increase in the thickness of the carotid artery intima tunic in the presence of atherosclerosis. Atherosclerosis intensified the inflammatory reaction and increased bone resorption in periapical lesions.

Phenotypic diversity of starch granules in cassava germplasm.

Demand for the development of cassava varieties with different native starches has guided the search for these characteristics in the germplasm of Manihot esculenta Crantz. Therefore, the objective of this study was to estimate the genetic diversity of cassava accessions for root and starch granule characteristics to guide the future industrial application of this species. Starches from 56 accessions were evaluated for the number of granules in 1 g of starch (NTG), area (AG, µm2), length (LG, µm), width (WG, µm), starch granule roundness (Round), dry matter content in the roots (DMC, %), pulp color (PulCo), and cyanogenic compounds (HCN). Images captured by light microscopy were used to determine the average phenotypic values, and these were further analyzed by principal component analysis (PCA) considering mixed data (quantitative and qualitative). Significant differences between the cassava accessions for all traits measured revealed wide variability in starch granule characteristics. Four diversity groups with better fitness for the classification of cassava accessions based on PulCo were identified, in comparison with HCN. Accessions with differential starch characteristics were identified, and crossings for the generation of segregating populations in order to obtain table and industry varieties have been proposed.

Fishing human monoclonal antibodies from a CHO cell supernatant with boronic acid magnetic particles.

In this work we have evaluated the potential of boronic acid functionalized magnetic particles for the one-step capture of a human monoclonal antibody (mAb) from a Chinese hamster ovary (CHO) cell culture supernatant. For comparison, Protein A coated magnetic particles were also used. The most important factor influencing the overall process yield and product purity in boronic acid particles was found to be the binding pH. Basic pH values promoted higher purities while resulting in decreased yields due to the competing effects of molecules such as glucose and lactate present in the cell culture supernatant. After optimization, the particles were successfully used in a multi-cycle purification process of the mAb from the CHO feedstock. Boronic acid particles were able to achieve an average overall yield of 86% with 88% removal of CHO host cell proteins (HCP) when the binding was performed at pH 7.4, while at pH 8.5 these values were 58% and 97%, respectively. In both cases, genomic DNA removal was in excess of 97%. Comparatively, Protein A particles recorded an average overall yield of 80% and an HCP removal greater than 99%. The adsorption of the mAb to the boronic acid particles was shown to be mediated by strong affinity interactions. Overall, boronic acid based purification processes can offer a cost-effective alternative to Protein A as the direct capturing step from the mammalian cell culture.

Identification of erythrobactin, a hydroxamate-type siderophore produced by Saccharopolyspora erythraea.

AIMS: To investigate the production of siderophores by Saccharopolyspora erythraea SGT2 and how this production is affected by the inoculum. METHODS AND RESULTS: When grown in a low-iron, chemically defined medium (CDM), the soil dwelling actinomycete S. erythraea secretes a substance that is reactive in the nonspecific chrome azurol S (CAS) assay. Importantly, the production of CAS-reactive substance is highly reduced upon the addition of 0.925 micromol l(-1) iron to the cultures and has a peak of production in the late-log to early stationary growth phase. In addition, the culture supernatants tested were negative in the Arnow and Rioux assays but positive in the Csáky procedure. Interestingly, we also found evidence that the production of this CAS-reactive substance in CDM was highly reduced, when inoculated with cells that had been previously grown to late-stationary phase. Conversely, inocula derived from late-log to early stationary cultures presented high levels of CAS activity. CONCLUSIONS: These results indicate that S. erythraea produces a hydroxamate-type siderophore that we have generically designated as erythrobactin. Additionally, the inocula growth stage plays a key role in siderophore production in S. erythraea. SIGNIFICANCE AND IMPACT OF THE STUDY: It is the first evidence for siderophore synthesis in S. erythraea and one of the first examples of non-polyketide secondary metabolite production by this organism.

Going down to the local: incorporating social organisation and political culture into assessments of decentralised health care.

The social organisation and political culture of the society in which an organisation is embedded can have major effects on the way in which organisational policy is implemented and on how that organisation functions. Research on health sector reforms has paid scant attention to this aspect. If the claims made for decentralised management in the health sector are to be evaluated seriously, it is critical to develop concepts and methods to evaluate not only the formal organisation and the outputs of the health system, but also the aspects of local social organisation and political culture within which that local health system is embedded that may mediate their relationship. The paper explores three cases of district health systems in Northeast Brazil in order to identify aspects of local social organisation and political culture that appear to influence the implementation of the reforms and thereby potentially impact upon the quality of the care provided. The results of the study indicate the importance that aspects of local social organisation and political culture may exert on the operations of a decentralised health system. Key aspects identified are: the space for autonomy; the space for local voice in political life; personalized and institutionalised influences on autonomy and local voice; differences of involvement of health staff with the district; different spaces of acceptable practice and accountability. These factors are seen to moderate the intent of the health reforms at all stages in their implementation. Three possibilities are discussed for the nature of the interaction in terms of cause and effect between the formal organisation of the health system and its local context. Seeing this relationship as one of a dialogue offers some cautious optimism for the potential of the reform agenda. The paper closes with suggestions on how to take this line of research forward.

A topographical display of epileptiform transients based on a statistical approach.

A method for the analysis of topographical relations of EEG epileptiform transients based on a statistical approach is described. The degree of time association of those transients recorded in pairs of derivations, or in 3 (triads), 4 (tetrads) or more derivations is estimated, using the chi2 statistic. Pairs of derivations which are found to be significantly associated are tabulated. This information is also graphically displayed on a computer drawing of the head (10-20 system). The same is done for triads, tetrads or higher order associations. The number of phase reversals between epileptiform transients found in significantly associated pairs of derivations is also computed. The resulting spatial displays enable a visualization of the location of the dominant epileptogenic area and of the spread of epileptiform activity over the head.

Effect of blood pH on anionic ferritin transport through rat aortic endothelium.

Acute lowering of blood pH between 7.4 and 6.9 in rats by ventilation with 10 or 20% CO2 does not increase the passage of ferritin molecules across the aortic endothelium. These results do not rule out alteration of endothelial permeability to anionic macromolecules in local circulatory disturbances when blood pH drops to levels much lower than 6.9.