Charge generation, charge transport, and residual charge in the electrospinning of polymers: A review of issues and complications

Electrospinning has become a widely implemented technique for the generation of nonwoven mats that are useful in tissue engineering and filter applications. The overriding factor that has contributed to the popularity of this method is the ease with which fibers with submicron diameters can be produced. Fibers on that size scale are comparable to protein filaments that are observed in the extracellular matrix. The apparatus and procedures for conducting electrospinning experiments are ostensibly simple. While it is rarely reported in the literature on this topic, any experience with this method of fiber spinning reveals substantial ambiguities in how the process can be controlled to generate reproducible results. The simplicity of the procedure belies the complexity of the physical processes that determine the electrospinning process dynamics. In this article, three process domains and the physical domain of charge interaction are identified as important in electrospinning: (a) creation of charge carriers, (b) charge transport, (c) residual charge. The initial event that enables electrospinning is the generation of region of excess charge in the fluid that is to be electrospun. The electrostatic forces that develop on this region of charged fluid in the presence of a high potential result in the ejection of a fluid jet that solidifies into the resulting fiber. The transport of charge from the charge solution to the grounded collection device produces some of the current which is observed. That transport can occur by the fluid jet and through the atmosphere surrounding the electrospinning apparatus. Charges that are created in the fluid that are not dissipated remain in the solidified fiber as residual charges. The physics of each of these domains in the electrospinning process is summarized in terms of the current understanding, and possible sources of ambiguity in the implementation of this technique are indicated. Directions for future research to further articulate the behavior of the electrospinning process are suggested. (C) 2012 American Institute of Physics. [doi: 10.1063/1.3682464]

Ordovician A-type granitoid magmatism on the Ceara Central Domain, Borborema Province, NE-Brazil

We present field relationships, major and trace element geochemistry and U-Pb SHRIMP and ID-TIMS geochronology of the A-type Ordovician Quintas pluton located in the Ceara Central Domain of the Borborema Province, in northeastern Brazil. This pluton presents a concentric geometry and is composed mainly of syenogranite, monzogranite, quartz syenite to quartz monzodiorite, monzogabbro and diorite. Its geochemical characteristics [SiO2 (52-70%), Na2O/K2O (1.55-0.65), Fe2O3/MgO (2.2-7.3), metaluminous to sligthly alkaline affinity, post-collisional type in (Y + Nb) x Rb diagram, and A-type affinity (Ga > 22 ppm, Nb > 20 ppm, Zn > 60 ppm), REE fractioned pattern with negative Eu anomaly] are coherent with post-collisional A(2)-type granitoids. However, the emplacement of this pluton is to some extent temporally associated with the deposition of the first strata of the Parnaiba intracratonic basin, attesting also to a purely anorogenic character (A(1)-type granitoid). The emplacement of this pluton is preceded by one of the largest known orogenesis of the planet (Neoproterozoic Pan-African/Brasiliano) and, if it is classified as an A(2)-type granitoid, it provides interesting constraints about how long can last A(2)-type magmatic activity after a major collisional episode, arguably triggered by disturbance of the underlying mantle, a topic extensively debated in the geoscience community. (C) 2011 Elsevier Ltd. All rights reserved.

Low resolution structural characterization of the Hsp70-interacting protein - Hip - from Leishmania braziliensis emphasizes its high asymmetry

The Hsp70 is an essential molecular chaperone in protein metabolism since it acts as a pivot with other molecular chaperone families. Several co-chaperones act as regulators of the Hsp70 action cycle, as for instance Hip (Hsp70-interacting protein). Hip is a tetratricopeptide repeat protein (TPR) that interacts with the ATPase domain in the Hsp70-ADP state, stabilizing it and preventing substrate dissociation. Molecular chaperones from protozoans, which can cause some neglected diseases, are poorly studied in terms of structure and function. Here, we investigated the structural features of Hip from the protozoa Leishmania braziliensis (LbHip), one of the causative agents of the leishmaniasis disease. LbHip was heterologously expressed and purified in the folded state, as attested by circular dichroism and intrinsic fluorescence emission techniques. LbHip forms an elongated dimer, as observed by analytical gel filtration chromatography, analytical ultracentrifugation and small angle X-ray scattering (SAXS). With the SAXS data a low resolution model was reconstructed, which shed light on the structure of this protein, emphasizing its elongated shape and suggesting its domain organization. We also investigated the chemical-induced unfolding behavior of LbHip and two transitions were observed. The first transition was related to the unfolding of the TPR domain of each protomer and the second transition of the dimer dissociation. Altogether. LbHip presents a similar structure to mammalian Hip, despite their low level of conservation, suggesting that this class of eukaryotic protein may use a similar mechanism of action. (C) 2012 Elsevier Inc. All rights reserved.

Identification of Regions Involved in Substrate Binding and Dimer Stabilization within the Central Domains of Yeast Hsp40 Sis1

Protein folding, refolding and degradation are essential for cellular life and are regulated by protein homeostatic processes such those that involve the molecular chaperone DnaK/Hsp70 and its co-chaperone DnaJ. Hsp70 action is initiated when proteins from the DnaJ family bind an unfolded protein for delivery purposes. In eukaryotes, the DnaJ family can be divided into two main groups, Type I and Type II, represented by yeast cytosolic Ydj1 and Sis1, respectively. Although sharing some unique features both members of the DnaJ family, Ydj1 and Sis1 are structurally and functionally distinct as deemed by previous studies, including the observation that their central domains carry the structural and functional information even in switched chimeras. In this study, we combined several biophysical tools for evaluating the stability of Sis1 and mutants that had the central domains (named Gly/Met rich domain and C-terminal Domain I) deleted or switched to those of Ydj1 to gain insight into the role of these regions in the structure and function of Sis1. The mutants retained some functions similar to full length wild-type Sis1, however they were defective in others. We found that: 1) Sis1 unfolds in at least two steps as follows: folded dimer to partially folded monomer and then to an unfolded monomer. 2) The Gly/Met rich domain had intrinsically disordered characteristics and its deletion had no effect on the conformational stability of the protein. 3) The deletion of the C-terminal Domain I perturbed the stability of the dimer. 4) Exchanging the central domains perturbed the conformational stability of the protein. Altogether, our results suggest the existence of two similar subdomains in the C-terminal domain of DnaJ that could be important for stabilizing each other in order to maintain a folded substrate-binding site as well as the dimeric state of the protein.

Analysis of MLL2 gene in the first Brazilian family with Kabuki syndrome

Most patients with Kabuki syndrome (KS) are the only person in their family with the condition. However, familial cases of KS have been described showing evidence that this syndrome can be inherited as a dominant trait with variable expressivity. We report on two related individuals with facial findings characteristic of KS. The proposita had arched eyebrows, long and upward slanting palpebral fissures, cleft lip and palate, retromicrognathia, brachydactyly of hands and feet, stubby fingers, nail hypoplasia, and prominent finger pads. Her mother had eyebrows with dispersed lateral half, long and upward slanting palpebral fissures, retrognathia, abnormal and posteriorly rotated ears, prominent finger pads, brachydactyly of feet, learning difficulties, and psychomotor development delay. DNA sequencing revealed a novel missense mutation in the MLL2 gene in both the proposita and her mother. The mutation (p.R5432Q) was found in the exon 51, within the SET domain of the gene, which confers methyltransferase activity on the protein. Therefore, the epigenetic and transcriptional regulatory properties of this protein may be altered and this suggests that the mutation is the cause of phenotype observed in both the patient and her mother. The clinical signs and the molecular evidence in this family further support the notion that KS is an autosomal dominant condition with variable expressivity. To our knowledge this is the first report of a Brazilian family with recurrence of this syndrome. (C) 2012 Wiley Periodicals, Inc.

Order-Disorder Transitions Govern Kinetic Cooperativity and Allostery of Monomeric Human Glucokinase

Glucokinase (GCK) catalyzes the rate-limiting step of glucose catabolism in the pancreas, where it functions as the body's principal glucose sensor. GCK dysfunction leads to several potentially fatal diseases including maturity-onset diabetes of the young type II (MODY-II) and persistent hypoglycemic hyperinsulinemia of infancy (PHHI). GCK maintains glucose homeostasis by displaying a sigmoidal kinetic response to increasing blood glucose levels. This positive cooperativity is unique because the enzyme functions exclusively as a monomer and possesses only a single glucose binding site. Despite nearly a half century of research, the mechanistic basis for GCK's homotropic allostery remains unresolved. Here we explain GCK cooperativity in terms of large-scale, glucose-mediated disorder-order transitions using 17 isotopically labeled isoleucine methyl groups and three tryptophan side chains as sensitive nuclear magnetic resonance (NMR) probes. We find that the small domain of unliganded GCK is intrinsically disordered and samples a broad conformational ensemble. We also demonstrate that small-molecule diabetes therapeutic agents and hyperinsulinemia-associated GCK mutations share a strikingly similar activation mechanism, characterized by a population shift toward a more narrow, well-ordered ensemble resembling the glucose-bound conformation. Our results support a model in which GCK generates its cooperative kinetic response at low glucose concentrations by using a millisecond disorder-order cycle of the small domain as a "time-delay loop," which is bypassed at high glucose concentrations, providing a unique mechanism to allosterically regulate the activity of human GCK under physiological conditions.

Stability-indicating methods for the enantioselective determination of dihydropyridines by high performance liquid chromatography and capillary electrophoresis

This paper presents simple, rapid, precise and accurate stability-indicating HPLC and CE methods, which were developed and validated for the determination of nitrendipine, nimodipine and nisoldipine. These drugs are calcium channel antagonists of the 1,4-dihydropyridine type which are used in the treatment of cardiovascular diseases. Experimental results showed a good linear correlation between the area and the concentration of drugs covering a relatively large domain of concentration in all cases. The linearity of the analytical procedures was in the range of 2.0-120.0 mu g mL-1 for nitrendipine, 1.0-100.0 mu g mL(-1) for nimodipine and 100.0-600.0 mu g mL(-1) for nisoldipine, the regression determination coefficient being higher than 0.99 in all cases. The proposed methods were found to have good precision and accuracy. The chemical stability of these drugs was determined under various conditions and the methods have shown adequate separation for their enantiomers and degradation products. In addition, degradation products produced as a result of stress studies did not interfere with the detection of the drugs' enantiomers and the assays can thus be considered stability-indicating.

ENERGY AND VOLUME OF VECTOR FIELDS ON SPHERICAL DOMAINS

We present a "boundary version" for theorems about minimality of volume and energy functionals on a spherical domain of an odd-dimensional Euclidean sphere.

Characterization of Stability Region for General Autonomous Nonlinear Dynamical Systems

The existing characterization of stability regions was developed under the assumption that limit sets on the stability boundary are exclusively composed of hyperbolic equilibrium points and closed orbits. The characterizations derived in this technical note are a generalization of existing results in the theory of stability regions. A characterization of the stability boundary of general autonomous nonlinear dynamical systems is developed under the assumption that limit sets on the stability boundary are composed of a countable number of disjoint and indecomposable components, which can be equilibrium points, closed orbits, quasi-periodic solutions and even chaotic invariant sets.

U-Pb zircon ages of orthogneisses and supracrustal rocks of the Cariris Velhos belt: Onset of Neoproterozoic rifting in the Borborema Province, NE Brazil

New geochronological and geochemical constraints on Precambrian sedimentary and volcanic successions exposed in the western part of the Central Domain of the Borborema Province, NE Brazil, indicate the presence of two distinct tectono-stratigraphic complexes: Riacho Gravata and Sao Caetano. Both complexes and associated orthogneisses are referred in the literature as the Cariris Velhos belt, having depositional, extrusive, or intrusive ages within the interval 985-913 Ma. The Riacho Gravata complex consists of bimodal (but mostly felsic) volcanic and volcanoclastic rocks, muscovite+/-graphite schists, quartzites, and marble with local occurrences of banded-iron-formation. The Sao Caetano complex mainly consists of metagreywackes, marbles, calc-silicate rocks, and rare meta-mafic rocks. Meta-mafic rocks from both complexes have geochemical signatures similar to those of continental flood basalts, with epsilon Nd (1.0 Ga) values ranging from -1.0 to -2.8. Felsic volcanic rocks from the Riacho Gravata complex show epsilon Nd (1.0 Ga) values ranging from -1.0 to -7.4 and geochemical signatures similar to A(2)-type granitoids. New SHRIMP U-Pb zircon data from felsic volcanic rocks within the Riacho Gravata complex yielded ages of 1091 +/- 13 Ma and 996 +/- 13 Ma. In contrast, meta-graywackes from the Sao Caetano complex show a maximum deposition age of ca. 806 Ma in the northern part and ca. 862 Ma in the southern part of the outcrop area. The orthogneisses show epsilon Nd (1.0 Ga) values ranging from 1.0 to -4.2 with U/Pb TIMS and SHRIMP ages ranging from 960 to 926 Ma and geochemical signatures of A(2)-type granitoids. The data reported in this paper suggest at least two periods of extension within the Central Domain of the Borborema Province, the first starts ca. 1091 Ma with magmatism and deposition, creating the Riacho Gravata basin and continued intrusion of A-type granites to 920 Ma. A second rift event, which reactivated old faults, generated a basin with a maximum deposition age of ca. 806 Ma. Furthermore, the oldest granitoids cutting these metasedimentary rocks have crystallization ages of ca. 600 Ma. This suggests that the second rift event could be early Brasiliano in age. The resulting Sao Caetano basin received detritus from a variety of sources, although detritus from the Riacho Gravata complex dominated. Deposition ages of the Riacho Gravata and the Sao Caetano complexes are coeval with deposits in other basins of the Borborema Province (Riacho do Tigre in the Central Domain; Macurure and Maranco in the Sergipano Belt of the Southern domain). The Macaubas Group from SE Brazil and its counterparts in Africa, the Zadanian and Mayumbian Groups, in the western edge of the Congo Craton are also coeval. Closure of the Riacho Gravata and Sao Caetano basins occurred during the Brasiliano convergence (705-600 Ma). During the last stage of convergence, ca. 612 Ma, pull-apart basins were created and filled; final basin closure took place 605-592 Ma, after deposition ceased. (C) 2011 Elsevier B.V. All rights reserved.

Effects of moisture content on structural and dielectric properties of cassava starch films

In this work, it was evaluated the effect of moisture content on the structural and dielectric properties of cassava starch films by means of Fourier Transform infrared spectroscopy (FTIR), impedancimetric, and gravimetric analysis. The film samples were equilibrated in hermetically sealed desiccators, containing different saturated salts in water in order to promote activity of water between 0.11 and 0.85. The position and amplitude of the peaks in the fingerprint region of the FTIR spectra were changed due to the modifications in the interactions between the polymeric chains and water molecules. These effects may be related to the formation of semi-crystalline regions in the film structure. The dielectric properties of the films were also strongly dependent on the moisture content, showing a non-linear and a linear region, which was attributed to the domain of bound and free water in the film, respectively. The gravimetric analyzes showed the typical sigmoidal behavior, attributed to the way the water interacts with the biopolymer. Finally, the flexibility of the films increased with water content increasing.

Loss of interest, depressed mood and impact on the quality of life: Cross-sectional survey

Abstract Background Depressive symptoms and chronic disease have adverse effects on patients' health-related quality of life (H-RQOL). However, little is known about this effect on H-RQOL when only the two core depressive symptoms - loss of interest and depressed mood - are considered. The objective of this study is to investigate H-RQOL in the presence of loss of interest and depressed mood at a general medical outpatient unit. Methods We evaluated 553 patients at their first attendance at a general medical outpatient unit of a teaching hospital. H-RQOL was assessed with the Medical Outcomes Study 36-item Short-Form Health Survey (SF-36). Depressed mood and loss of interest were assessed by the Primary Care Evaluation of Mental Disorders (PRIME-MD)-Patient Questionnaire. A physician performed the diagnosis of chronic diseases by clinical judgment and classified them in 13 possible pre-defined categories. We used multiple linear regression to investigate associations between each domain of H-RQOL and our two core depression symptoms. The presence of chronic diseases and demographic variables were included in the models as covariates. Results Among the 553 patients, 70.5% were women with a mean age of 41.0 years (range 18-85, SD ± 15.4). Loss of interest was reported by 54.6%, and depressed mood by 59.7% of the patients. At least one chronic disease was diagnosed in 59.5% of patients; cardiovascular disease was the most prevalent, affecting 20.6% of our patients. Loss of interest and depressed mood was significantly associated with decreased scores in all domains of H-RQOL after adjustment for possible confounders. The presence of any chronic disease was associated with a decrease in the domain of vitality. The analysis of each individual chronic disease category revealed that no category was associated with a decrease in more than one domain of H-RQOL. Conclusion Loss of interest and depressed mood were associated with significant decreases in H-RQOL. We recommend these simple tests for screening in general practice.

Genomic analysis of ERVWE2 locus in patients with multiple sclerosis: absence of genetic association but potential role of human endogenous retrovirus type W elements in molecular mimicry with myelin antigen

Human endogenous retroviruses (HERVs) arise from ancient infections of the host germline cells by exogenous retroviruses, constituting 8% of the human genome. Elevated level of envelope transcripts from HERVs-W has been detected in CSF, plasma and brain tissues from patients with Multiple Sclerosis (MS), most of them from Xq22.3, 15q21.3, and 6q21 chromosomes. However, since the locus Xq22.3 (ERVWE2) lack the 5' LTR promoter and the putative protein should be truncated due to a stop codon, we investigated the ERVWE2 genomic loci from 84 individuals, including MS patients with active HERV-W expression detected in PBMC. In addition, an automated search for promoter sequences in 20 kb nearby region of ERVWE2 reference sequence was performed. Several putative binding sites for cellular cofactors and enhancers were found, suggesting that transcription may occur via alternative promoters. However, ERVWE2 DNA sequencing of MS and healthy individuals revealed that all of them harbor a stop codon at site 39, undermining the expression of a full-length protein. Finally, since plaque formation in central nervous system (CNS) of MS patients is attributed to immunological mechanisms triggered by autoimmune attack against myelin, we also investigated the level of similarity between envelope protein and myelin oligodendrocyte glycoprotein (MOG). Comparison of the MOG to the envelope identified five retroviral regions similar to the Ig-like domain of MOG. Interestingly, one of them includes T and B cell epitopes, capable to induce T effector functions and circulating Abs in rats. In sum, although no DNA substitutions that would link ERVWE2 to the MS pathogeny was found, the similarity between the envelope protein to MOG extends the idea that ERVEW2 may be involved on the immunopathogenesis of MS, maybe facilitating the MOG recognizing by the immune system. Although awaiting experimental evidences, the data presented here may expand the scope of the endogenous retroviruses involvement on MS pathogenesis

The importance of vertical and horizontal dimensions of the sediment matrix in structuring nematodes across spatial scales

Intensive surveys have been conducted to unravel spatial patterns of benthic infauna communities. Although it has been recognized that benthic organisms are spatially structured along the horizontal and vertical dimensions of the sediment, little is known on how these two dimensions interact with each other. In this study we investigated the interdependence between the vertical and horizontal dimensions in structuring marine nematodes assemblages. We tested whether the similarity in nematode species composition along the horizontal dimension was dependent on the vertical layer of the sediment. To test this hypothesis, three-cm interval sediment samples (15 cm depth) were taken independently from two bedforms in three estuaries. Results indicated that assemblages living in the top layers are more abundant, species rich and less variable, in terms of species presence/absence and relative abundances, than assemblages living in the deeper layers. Results showed that redox potential explained the greatest amount (12%) of variability in species composition, more than depth or particle size. The fauna inhabiting the more oxygenated layers were more homogeneous across the horizontal scales than those from the reduced layers. In contrast to previous studies, which suggested that reduced layers are characterized by a specific set of tolerant species, the present study showed that species assemblages in the deeper layers are more causal (characterized mainly by vagrant species). The proposed mechanism is that at the superficial oxygenated layers, species have higher chances of being resuspended and displaced over longer distances by passive transport, while at the deeper anoxic layers they are restricted to active dispersal from the above and nearby sediments. Such restriction in the dispersal potential together with the unfavorable environmental conditions leads to randomness in the presence of species resulting in the high variability between assemblages along the horizontal dimension.

Factor Xa activation of factor V is of paramount importance in initiating the coagulation system: lessons from a tick salivary protein

BACKGROUND: Generation of active procoagulant cofactor factor Va (FVa) and its subsequent association with the enzyme activated factor X (FXa) to form the prothrombinase complex is a pivotal initial event in blood coagulation and has been the subject of investigative effort, speculation, and controversy. The current paradigm assumes that FV activation is initiated by limited proteolysis by traces of (meizo) thrombin. METHODS AND RESULTS: Recombinant tick salivary protein TIX-5 was produced and anticoagulant properties were studied with the use of plasma, whole blood, and purified systems. Here, we report that TIX-5 specifically inhibits FXa-mediated FV activation involving the B domain of FV and show that FXa activation of FV is pivotal for plasma and blood clotting. Accordingly, tick feeding is impaired on TIX-5 immune rabbits, displaying the in vivo importance of TIX-5. CONCLUSIONS: Our data elucidate a unique molecular mechanism by which ticks inhibit the host's coagulation system. From our data, we propose a revised blood coagulation scheme in which direct FXa-mediated FV activation occurs in the initiation phase during which thrombin-mediated FV activation is restrained by fibrinogen and inhibitors.