Composition of a protein-like fluorophore of dissolved organic matter in coastal wetland and estuarine ecosystems

This study demonstrates the compositional heterogeneity of a protein-like fluorescence emission signal (T-peak; excitation/emission maximum at 280/325 nm) of dissolved organic matter (DOM) samples collected from subtropical river and estuarine environments. Natural water samples were collected from the Florida Coastal Everglades ecosystem. The samples were ultrafiltered and excitation–emission fluorescence matrices were obtained. The T-peak intensity correlated positively with N concentration of the ultrafiltered DOM solution (UDON), although, the low correlation coefficient (r2=0.140, p<0.05) suggested the coexistence of proteins with other classes of compounds in the T-peak. As such, the T-peak was unbundled on size exclusion chromatography. The elution curves showed that the T-peak was composed of two compounds with distinct molecular weights (MW) with nominal MWs of about >5×104 (T1) and ∼7.6×103 (T2) and with varying relative abundance among samples. The T1-peak intensity correlated strongly with [UDON] (r2=0.516, p<0.001), while T2-peak did not, which suggested that the T-peak is composed of a mixture of compounds with different chemical structures and ecological roles, namely proteinaceous materials and presumably phenolic moieties in humic-like substances. Natural source of the latter may include polyphenols leached from senescent plant materials, which are important precursors of humic substances. This idea is supported by the fact that polyphenols, such as gallic acid, an important constituent of hydrolysable tannins, and condensed tannins extracted from red mangrove (Rhizophora mangle) leaves exhibited the fluorescence peak in the close vicinity of the T-peak (260/346 and 275/313 nm, respectively). Based on this study the application of the T-peak as a proxy for [DON] in natural waters may have limitations in coastal zones with significant terrestrial DOM input.

Molecular characterization of complete and incomplete immunoglobulin heavy chain gene rearrangements in hairy cell leukemia.

PURPOSE: We analyzed patients with hairy cell leukemia (HCL) to achieve a better understanding of the differentiation stage reached by HCL cells and to define the key role of the diversification of cell surface makers, especially CD25 expression. PATIENTS AND METHODS: We analyzed 38 previously untreated patients with HCL to characterize their complete (VDJ(H)) and incomplete (DJ(H)) immunoglobulin (Ig) heavy chain (IgH) rearrangements, including somatic hypermutation pattern and gene segment use. RESULTS: A correlation between immunophenotypic profile and molecular data was seen. All 38 cases showed monoclonal amplifications: VDJ(H) in 97%, DJ(H) in 42%, and both in 39%. Segments from the D(H)3 family were used more in complete compared with incomplete rearrangements (45% vs. 12%; P <.005). Furthermore, comparison between molecular and immunophenotypic characteristics disclosed differences in the expression of CD25 antigen; CD25(-) cases, a phenotype associated with HCL variant, showed complete homology to the germline in 3 of 5 cases (60%), whereas this characteristic was never observed in CD25(+) cases (P <.005). Moreover, V(H)4-34, V(H)1-08, and J(H)3 segments appeared in 2, 1, and 2 CD25(-) cases, respectively, whereas they were absent in all CD25(+) cases. CONCLUSION: These results support that HCL is a heterogeneous entity including subgroups with different molecular characteristics, which reinforces the need for additional studies with a larger number of patients to clarify the real role of gene rearrangements in HCL.

The effects of PVP(FE(III)) catalyst and polymer molecular weight on gene delivery via biodegradable crosslinked polyethylenimine

Human gene therapy has faced many setbacks due to the immunogenicity and oncogenity of viruses. Safe and efficient alternative gene delivery vehicles are needed to implement gene therapy in clinical practice. Polymeric vectors are an attractive option due to their availability, simple chemistry, and low toxicity and immunogenicity. Our group has previously reported biodegradable polyethylenimines (PEI) that show high transfection efficiency and low toxicity by cross-linking 800 Da PEI with diacrylate cross-linkers using Michael addition. However, the synthesis was difficult to control, inconsistent, and resulted in polymers with a narrow range of molecular weights. In the present work, we utilized a heterogenous PVP(Fe(III)) catalyst to provide a more controllable PEI crosslinking reaction and wider range of biodegradable PEIs. The biodegradable PEIs reported here have molecular weights ranging from 1.2 kDa to 48 kDa, are nontoxic in MDA-MB-231 cells, and show low toxicity in HeLa cells. At their respective optimal polymer:DNA ratios, these biodegradable PEIs demonstrated about 2-5-fold higher transfection efficiency and 2-7-fold higher cellular uptake, compared unmodified 25 kDa PEI. The biodegradable PEIs show similar DNA condensation properties as unmodified PEI but more readily unpackage DNA, based on ethidium bromide exclusion and heparan sulfate competitive displacement assays, which could contribute to their improved transfection efficiency. Overall, the synthesis reported here provides a more robust, controlled reaction to produce cross-linked biodegradable PEIs that show enhanced gene delivery, low toxicity, and high cellular uptake and can potentially be used for future in vivo studies.

Avaliação da hidrólise alcalina da quitina e elaboração de filmes de quitosana para aplicação na adsorção do corante têxtil reativo preto 5

Nesta pesquisa, diferentes amostras de quitosana foram produzidas por diferentes condições de hidrólise alcalina da quitina. A partir das amostras de quitosana foram produzidos filmes,sendo estes aplicados na adsorção do corante têxtil reativo preto 5 e os resultados foram comparados com os dos seus respectivos pós. Os valores das massas molares da quitosana aumentaram em função do aumento do diâmetro da quitina e diminuíram com o aumento da relação de solução NaOH:quitina, da concentração de NaOH e tempo de reação, e ficaram na faixa de 100 a 200 kDa. Um comportamento inverso foi observado para o grau de desacetilação da quitosana, e seus valores variaram de 65 a 95%. Quanto aos filmes biopoliméricos elaborados, os que apresentaram melhores valores quanto as suas propriedades mecânicas e de permeabilidade ao vapor de água foram os filmes produzidos com quitosana de mais elevada massa molar e menor grau de desacetilação. A fim de avaliar o comportamento dos filmes em processos de adsorção, estes foram aplicados na remoção do corante reativo preto 5 (RB5) em diferentes condições de pH (4, 6 e 8). Após, foram escolhidos quatro filmes de quitosana (FQ), com diferentes graus de desacetilação e massas molares, que foram comparados com as quitosanas na forma de pó (PQ) no estudo de adsorção. Este foi realizado sob diversas condições experimentais (pH, temperatura e taxa de agitação) através das isotermas de equilíbrio, da termodinâmica e da cinética. Análises de interação e ciclos de adsorção-dessorção também foram realizados. Verificou-se que PQ e FQ com grau de desacetilação de 95% e massa molar de 100 kDa foram os adsorvente mais adequados, apresentando mais de 99% de remoção do corante RB5 em pH 4,0. Para ambos, PQ e FQ, o modelo de Langmuir foi o mais adequado para representar os dados de equilíbrio. As capacidades máximas de adsorção foram 654,3 e 589,5 mg g-1 para PQ e FQ, respectivamente, obtidos a 298 K. O processo de adsorção foi espontâneo, favorável e exotérmico. A adsorção de RB5 para PQ e FQ seguiu o modelo cinético de Elovich,e ocorreram interações eletrostáticas do PQ-RB5 e do FQ-RB5. Os filmes de quitosana foram reutilizados três vezes, enquanto que a quitosana em pó não pode ser reutilizada.

Unravelling the nanostructure of strawberry fruit pectins by atomic force microscopy

Atomic force microscopy (AFM) allows the analysis of individual polymers at nanostructural level with a minimal sample preparation. This technique has been used to analyse the pectin disassembly process during the ripening and postharvest storage of several fleshy fruits. In general, pectins analysed by AFM are usually visualized as isolated chains, unbranched or with a low number of branchs and, occasionally, as large aggregates. However, the exact nature of these structures is unknown. It has been suggested that pectin aggregates represent a mixture of rhamnonogalacturonan I and homogalacturonan, while isolated chains and their branches are mainly composed by polygalacturonic acid. In order to gain insight into the nature of these structures, sodium carbonate soluble pectins from ripe strawberry (Fragaria x ananassa, Duch.) fruits were subjected to enzymatic digestion with endo-Polygalacturonase M2 from Aspergillus aculeatus, and the samples visualized by AFM at different time intervals. Pectins isolated from control, non-transformed plants, and two transgenic genotypes with low level of expression of ripening-induced pectinase genes encoding a polygalacturonase (APG) or a pectate lyase (APEL) were also included in this study. Before digestion, isolated pectin chains from control were shorter than those from transgenic fruits, showing number-average (LN) contour length values of 73.2 nm vs. 95.9 nm and 91.4 nm in APG and APEL, respectively. The percentage of branched polymers was significantly higher in APG polyuronides than in the remaining genotypes, 33% in APG vs. 6% in control and APEL. As a result of the endo-PG treatment, a gradual decrease in the main backbone length of isolated chains was observed in the three samples. The minimum LN value was reached after 8 h of digestion, being similar in the three genotypes, 22 nm. By contrast, the branches were not visible after 1.5-2 h of digestion. LN values were plotted against digestion time and the data fitted to a first-order exponential decay curve, obtaining R2 values higher than 0.9. The half digestion time calculated with these equations were similar for control and APG pectins, 1.7 h, but significantly higher in APEL, 2.5 h, indicating that these polymer chains were more resistant to endo-PG digestion. Regarding the pectin aggregates, their volumes were estimated and used to calculate LN molecular weights. Before digestion, control and APEL samples showed complexes of similar molecular weights, 1722 kDa, and slightly higher than those observed in APG samples. After endo-PG digestion, size of complexes diminished significantly, reaching similar values in the three pectin samples, around 650 kDa. These results suggest that isolated polymer chains visualized by AFM are formed by a HG domain linked to a shorter polymer resistant to endo-PG digestion, maybe xylogalacturonan or RG-I. The silencing of the pectate lyase gene slightly modified the structure and/or chemical composition of polymer chains making these polyuronides more resistant to enzymatic degradation. Similarly, polygalacturonic acid is one of the main component of the aggregates.

Porous self-protonating spiropyran-based NIPAAm gels with improved reswelling kinetics

The final publication is available at Springer via http://dx.doi.org/[10.1007/s10853-015-9458-2]

Raman Spectroscopy and DFT calculations for the Electronic Structure Characterization of Conjugated Polymers

In the last three decades, there has been a broad academic and industrial interest in conjugated polymers as semiconducting materials for organic electronics. Their applications in polymer light-emitting diodes (PLEDs), polymer solar cells (PSCs), and organic field-effect transistors (OFETs) offer opportunities for the resolution of energy issues as well as the development of display and information technologies1. Conjugated polymers provide several advantages including low cost, light weight, good flexibility, as well as solubility which make them readily processed and easily printed, removing the conventional photolithography for patterning2. A large library of polymer semiconductors have been synthesized and investigated with different building blocks, such as acenes or thiophene and derivatives, which have been employed to design new materials according to individual demands for specific applications. To design ideal conjugated polymers for specific applications, some general principles should be taken into account, including (i) side chains (ii) molecular weights, (iii) band gap and HOMO and LUMO energy levels, and (iv) suited morphology.3-6 The aim of this study is to elucidate the impact that substitution exerts on the molecular and electronic structure of π-conjugated polymers with outstanding performances in organic electronic devices. Different configurations of the π-conjugated backbones are analyzed: (i) donor-acceptor configuration, (ii) 1D lineal or 2D branched conjugated backbones, and (iii) encapsulated polymers (see Figure 1). Our combined vibrational spectroscopy and DFT study shows that small changes in the substitution pattern and in the molecular configuration have a strong impact on the electronic characteristics of these polymers. We hope this study can advance useful structure-property relationships of conjugated polymers and guide the design of new materials for organic electronic applications.

Modelling the influence of diradical character and aggregation on conjugated molecular materials

Molecular materials are made by the assembly of specifically designed molecules to obtain bulk structures with desired solid-state properties, enabling the development of materials with tunable chemical and physical properties. These properties result from the interplay of intra-molecular constituents and weak intermolecular interactions. Thus, small changes in individual molecular and electronic structure can substantially change the properties of the material in bulk. The purpose of this dissertation is, thus, to discuss and to contribute to the structure-property relationships governing the electronic, optical and charge transport properties of organic molecular materials through theoretical and computational studies. In particular, the main focus is on the interplay of intra-molecular properties and inter-molecular interactions in organic molecular materials. In my three-years of research activity, I have focused on three major areas: 1) the investigation of isolated-molecule properties for the class of conjugated chromophores displaying diradical character which are building blocks for promising functional materials; 2) the determination of intra- and intermolecular parameters governing charge transport in molecular materials and, 3) the development and application of diabatization procedures for the analysis of exciton states in molecular aggregates. The properties of diradicaloids are extensively studied both regarding their ground state (diradical character, aromatic vs quinoidal structures, spin dynamics, etc.) and the low-lying singlet excited states including the elusive double-exciton state. The efficiency of charge transport, for specific classes of organic semiconductors (including diradicaloids), is investigated by combining the effects of intra-molecular reorganization energy, inter-molecular electronic coupling and crystal packing. Finally, protocols aimed at unravelling the nature of exciton states are introduced and applied to different molecular aggregates. The role of intermolecular interactions and charge transfer contributions in determining the exciton state character and in modulating the H- to J- aggregation is also highlighted.

Tempo de fornecimento da dieta pré-inicial para frangos de corte com diferentes pesos ao alojamento

Avaliaram-se os efeitos do fornecimento da dieta pré-inicial (DPI) e do peso ao alojamento sobre o desempenho e as características de carcaça de frangos de corte. Utilizaram-se 960 frangos machos, linhagem comercial Cobb-500, alojados em 32 boxes, em um delineamento inteiramente casualizado em esquema fatorial 2 x 4, composto de dois pesos ao alojamento (42 e 46 g) e quatro períodos de fornecimento da dieta pré-inicial (0, 1 a 7, 1 a 10 e 1 a 14 dias de idade), durante 49 dias. Os programas alimentares compreenderam três ou quatro fases (pré-inicial e/ou inicial; engorda; final) e as dietas fornecidas eram à base de milho e farelo de soja. De modo geral, aves mais pesadas ao alojamento apresentaram, ao longo de todo o período experimental, maior consumo de ração (CR) e ganho de peso (GP). Não foram observadas diferenças significativas na conversão alimentar (CA) dos animais. Considerando todo o período experimental, as aves alimentadas com a dieta pré-inicial durante sete dias apresentaram maior consumo de ração e ganho de peso, porém, menor rendimento de carcaça. O peso ao alojamento determina o desempenho e o oferecimento da dieta pré-inicial durante sete dias beneficia o desempenho das aves.

Qualidade da carne de suínos de três linhagens genéticas comerciais em diferentes pesos de abate

O objetivo deste trabalho foi determinar as características de qualidade da carne de suínos de diferentes linhagens genéticas, em diferentes pesos de abate. Neste estudo foram utilizados 88 suínos por linhagem, fêmeas e machos castrados, com idade e peso médio iniciais de 74 dias e 30kg, respectivamente, pertencentes a três linhagens genéticas distintas, designadas de AgroceresPic, Dalland e Seghers. A etapa experimental foi dividida em quatro fases (Crescimento I, Crescimento II, Terminação I e Terminação II). Ao final de cada etapa, foram abatidos 60 animais (10 por linhagem/sexo), para as análises de qualidade da carne. O pH e a temperatura foram determinados a 1 e 24 horas post mortem. Foram retiradas amostras para as determinações de cor, perda de água por exsudação (PAE), perda de água por cocção (PAC) e força de Cisalhamento. As amostras provenientes das carcaças dos animais da linhagem AgroceresPic e Dalland apresentaram valores médios de PAE superiores aos da linhagem Seghers. Os valores médios de PAC foram diferentes entre as diferentes fases. Na fase Crescimento II, as carnes provenientes das carcaças das fêmeas Dalland foram mais duras, ou seja, com maiores valores de força de cisalhamento, que as carnes dos machos. Entretanto, na linhagem Seghers, as carnes dos machos foram mais duras. Observou-se diferença de L* entre os sexos da linhagem AgroceresPic nas fases Terminação I e II e, na linhagem Dalland, na Terminação II. As linhagens Dalland e Seghers apresentaram carnes com resultados superiores de a*, parâmetro característico da cor vermelha (a*>0). Pode-se concluir, portanto, que as características de qualidade da carne de suínos (pH, perda de água por exsudação, cor e maciez) podem variar entre grupos genéticos, entre sexos e entre diferentes pesos ao abate.

Características de carcaça de suínos de três linhagens genéticas em diferentes idades ao abate

O objetivo deste trabalho foi estudar as características da carcaça de suínos de diferentes linhagens genéticas, em diferentes idades ao abate. Foram utilizados 88 suínos por linhagem, fêmeas e machos castrados, com idade e peso médio iniciais de 74 dias e 30kg, respectivamente, pertencentes a três linhagens genéticas distintas, designadas de AgroceresPic, Dalland e Seghers. A etapa experimental foi dividida em quatro fases (Crescimento I, Crescimento II, Terminação I e Terminação II). Ao final de cada etapa, foram abatidos 60 animais (10 por linhagem/sexo), para as caracterizações: Peso (PCQ) e Rendimento de Carcaça Quente (RCQ), Área de Olho de Lombo (AOL) e Espessura de Toucinho (ET). Na análise dos resultados foi utilizado um delineamento inteiramente casualizado com desdobramento dos graus de liberdade em esquema fatorial 4 x 3 x 2, sendo quatro idades ao abate (90, 119, 150 e 186 dias), três linhagens (AgroceresPic, Dalland e Seghers) e dois sexos (fêmea e macho castrado), com 10 repetições por tratamento, sendo utilizado o pacote PROC MIXED do Softwear SAS. Os valores médios de peso vivo apresentaram diferença entre as linhagens e interação entre fase e linhagem (P<0,05). Os valores médios das demais variáveis estudadas apresentaram diferença e a interação (P<0,05) entre fase, linhagem e sexo, sendo que de maneira geral as principais diferenças ocorreram a partir da fase de Terminação I, em que as fêmeas das linhagens AgroceresPic e Dalland apresentaram melhores resultados (P<0,05) de RCQ (80,4 e 80,7%, respectivamente) em comparação com os machos (78,8 e 78,7%, respectivamente) e além disso as fêmeas Dalland apresentaram valores superiores (P<0,01) de AOL e ET (45,7cm² e 11,4mm x 38,3cm² e 18,3mm). Conclui-se que as linhagens genéticas avaliadas apresentaram características de carcaça muito interessantes para o mercado atual e que as fêmeas suínas podem ser utilizadas em programas que visem o abate de animais mais pesados.

Influência das características das substâncias húmicas aquáticas na eficiência da coagulação com o cloreto férrico

The aim of this study was to verify the influence of the apparent molecular size of aquatic humic substances on the effectiveness of coagulation with ferric chloride. Coagulation-filtration tests using jar test and bench-scale sand filters were carried out on samples of water with true color of approximately 100 Hazen units, prepared with aquatic humic substances of different molecular sizes (F1: < 0.45 µm, F2: 100 kDa - 0.45 µm, F3: 30 - 100 kDa and F4': < 30 kDa). For the water samples with lower apparent molecular size fractions, greater dosages of coagulant was needed to remove the color around 5.0 Hanzen units, mainly because these water samples contain higher concentrations of fulvic acids, which exhibited a larger number of negatively-charged groups.

Prognostic index expression of cyclin-D1, cerbB-2 and VEGF: metastases vs corresponding primary cancers and metastatic vs non-metastatic adenocarcinomas

The prognostic relevance of different molecular markers in lung cancer is a crucial issue still worth investigating, and the specimens collected and analyzed represent a valuable source of material. Cyclin-D1, c-erbB-2 and vascular endothelial growth factor (VEGF) have shown to be promising as prognosticators in human cancer. In this study, we sought to examine the importance of Cyclin-D1, c-erbB-2 and VEGF, and to study the quantitative relationship among these factors and disease progression in metastases vs corresponding primary cancer, and metastatic vs non metastatic cancers. Material and Methods: We used immunohistochemistry and morphometric analysis to evaluate the amount of tumour staining for Cyclin-D1, c-erbB-2 and VEGF in 52 patients with surgically excised ademocarcinoma of the lung, and the outcome for our study was survival time until death from hematogenic metastases. Results: Metastasis presented lower c-erbB-2 expression than corresponding primary cancers (p=0.02). Cyclin-D1 and VEGF expression were also lower in metastases than in corresponding primary cancers, but this difference did not achieve statistical significance. Non-metastatic cancers also presented significantly lower Cyclin-D1 and c-erbB-2 expression than metastatic cancers (p<0.01 and p<0.01, respectively). Equally significant was the difference between higher c-erbB-2 expression by metastatic cancers compared to non-metastatic cancers (p=0.02). Considering survival in Kaplan-Maier analysis, Cyclin-D1 (p=0.04), c-erbB-2 (p=0.04) and VEGF (p<0.01) were important predictors of survival in metastatic cancers.

Solution Polymerization of N-vinylcaprolactam in 1,4-dioxane. Kinetic Dependence on Temperature, Monomer, and Initiator Concentrations

The kinetics of the solution free radical polymerization of N-vinylcaprolactam, in 1,4-dioxane and under various polymerization conditions was studied. Azobisisobutyronitrile and 3-mercaptopropionic acid were used as initiator and as chain transfer agent (CTA), respectively. The influence of monomer and initiator concentrations and polymerization temperature on the rate of polymerizations (R(p)) was investigated. In general, high conversions were obtained. The order with respect to initiator was consistent with the classical kinetic rate equation, while the order with respect to the monomer was greater than unity. The overall activation energy of 53.6 kJ mol(-1) was obtained in the temperature range 60-80 degrees C. The decreasing of the absolute molecular weights when increasing the CIA concentration was confirmed by GPC/SEC/LALS analyses. It was confirmed by UV-visible analyses the effect of molecular weights on the lower critical solution temperature of the polymers. It was also verified that the addition of the CTA influenced the kinetic of the polymerizations. (C) 2010 Wiley Periodicals, Inc. J Appl Polym Sci 118: 229-240, 2010

Linear viscoelasticity of styrenic block copolymers-clay nanocomposites

In this work, the rheological behavior of block copolymers with different morphologies (lamellar, cylindrical, spherical, and disordered) and their clay-containing nanocomposites was studied using small amplitude oscillatory shear. The copolymers studied were one asymmetric starblock styrene-butadiene-styrene copolymer and four styrene-ethylene/butylenes-styrene copolymers with different molecular architectures, one of them being modified with maleic anhydride. The nanocomposites of those copolymers were prepared by adding organophilic clay using three different preparation techniques: melt mixing, solution casting, and a hybrid melt mixing-solution technique. The nanocomposites were characterized by X-ray diffraction and transmission electron microscopy, and their viscoelastic properties were evaluated and compared to the ones of the pure copolymers. The influence of copolymer morphology and presence of clay on the storage modulus (G`) curves was studied by the evaluation of the changes in the low frequency slope of log G` x log omega (omega: frequency) curves upon variation of temperature and clay addition. This slope may be related to the degree of liquid- or solid-like behavior of a material. It was observed that at temperatures corresponding to the ordered state, the rheological behavior of the nanocomposites depended mainly on the viscoelasticity of each type of ordered phase and the variation of the slope due to the addition of clay was small. For temperatures corresponding to the disordered state, however, the rheological behavior of the copolymer nanocomposites was dictated mostly by the degree of clay dispersion: When the clay was well dispersed, a strong solid-like behavior corresponding to large G` slope variations was observed.