Microtubule-associated protein 1B binds glyceraldehyde-3-phosphate dehydrogenase.

Microtubule-associated protein 1B, MAP1B, is a major cytoskeletal protein during brain development and one of the largest brain MAPs associated with microtubules and microfilaments. Here, we identified several proteins that bind to MAP1B via immunoprecipitation with a MAP1B-specific antibody, by one and two-dimensional gel electrophoresis and subsequent mass spectrometry identification of precipitated proteins. In addition to tubulin and actin, a variety of proteins were identified. Among these proteins were glyceraldehyde-3-phosphate dehydrogenase (GAPDH), heat shock protein 8, dihydropyrimidinase related proteins 2 and 3, protein-L-isoaspartate O-methyltransferase, beta-spectrin, and clathrin protein MKIAA0034, linking either directly or indirectly to MAP1B. In particular, GAPDH, a key glycolytic enzyme, was bound in large quantity to the heavy chain of MAP1B in adult brain tissue. In vitro binding studies confirmed a direct binding of GAPDH to MAP1B. In PC12 cells, GAPDH was found in cytoplasm and nuclei and partially co-localized with MAP1B. It disappeared from the cytoplasm under oxidative stress or after a disruption of cytoskeletal elements after colcemid or cytochalasin exposure. GAPDH may be essential in the local energy provision of cytoskeletal structures and MAP1B may help to keep this key enzyme close to the cytoskeleton.

In-vivo performance of high-density collagen gel tubes for urethral regeneration in a rabbit model.

Congenital malformations or injuries of the urethra can be treated using existing autologous tissue, but these procedures are sometimes associated with severe complications. Therefore, tissue engineering may be advantageous for generating urethral grafts. We evaluated engineered high-density collagen gel tubes as urethral grafts in 16 male New Zealand white rabbits. The constructs were either acellular or seeded with autologous smooth muscle cells, isolated from an open bladder biopsy. After the formation of a urethral defect by excision, the tissue-engineered grafts were interposed between the remaining urethral ends. No catheter was placed postoperatively. The animals were evaluated at 1 or 3 months by contrast urethrography and histological examination. Comparing the graft caliber to the control urethra at 3 months, a larger caliber was found in the cell-seeded grafts (96.6% of the normal caliber) than in the acellular grafts (42.3%). Histology of acellular and cell-seeded grafts did not show any sign of inflammation, and spontaneous regrowth of urothelium could be demonstrated in all grafts. Urethral fistulae, sometimes associated with stenosis, were observed, which might be prevented by urethral catheter application. High-density collagen gel tubes may be clinically useful as an effective treatment of congenital and acquired urethral pathologies.

Variabilidade genética em cultivares de soja determinada com marcadores microssatélites em gel de agarose

O objetivo deste trabalho foi avaliar a variabilidade genética de cultivares de soja com marcadores microssatélites selecionados e caracterizados quanto à informatividade para uso em gel de agarose. O DNA de 23 cultivares de soja foi amplificado com 283 marcadores microssatélites em gel de agarose a 3%. Posteriormente, 53 marcadores que apresentaram polimorfismo facilmente detectável nos géis de agarose foram utilizados na caracterização de 53 cultivares. Nessas cultivares foram detectados 124 alelos, com média de 2,34 alelos por loco, e os valores de conteúdo de informação polimórfica variaram entre 0,16 e 0,66, com média de 0,47. As frequências alélicas variaram de 0,02 a 0,91, com média de 0,43. A distância genética calculada variou de 0,02 a 0,73, com média de 0,47. A menor distância observada foi entre as cultivares CD201 e CD208, e a maior distância entre CD210 e BRSMT Uirapuru. Os marcadores utilizados possibilitaram a identificação das 53 cultivares avaliadas. Os locos microssatélites de soja, avaliados em gel de agarose, apresentam elevada informatividade. É possível detectar variabilidade significativa no germoplasma brasileiro de soja avaliado, mesmo entre cultivares elite, quando se usa marcadores moleculares microssatélites selecionados por sua informatividade.

Sedimentation of macroscopic rigid knots and its relation to gel electrophoretic mobility of DNA knots.

We address the general question of the extent to which the hydrodynamic behaviour of microscopic freely fluctuating objects can be reproduced by macrosopic rigid objects. In particular, we compare the sedimentation speeds of knotted DNA molecules undergoing gel electrophoresis to the sedimentation speeds of rigid stereolithographic models of ideal knots in both water and silicon oil. We find that the sedimentation speeds grow roughly linearly with the average crossing number of the ideal knot configurations, and that the correlation is stronger within classes of knots. This is consistent with previous observations with DNA knots in gel electrophoresis.

Investigations of a thermosensitive gel to temporarily occlude crural arteries in femoro-distal bypass surgery.

OBJECTIVES: Long occlusions in calcified crural arteries are a major cause of endovascular technical failure in patients with critical limb ischaemia. Therefore, distal bypasses are mainly performed in patients with heavily calcified arteries and with consequently delicate clamping. A new reverse thermosensitive polymer (RTP) is an alternative option to occlude target vessels. The aim of the study is to report our technical experience with RTP and to assess its safety and efficiency to temporarily occlude small calcified arteries during anastomosis time. METHODS: Between July 2010 and December 2011, we used RTP to occlude crural arteries in 20 consecutive patients with 20 venous distal bypasses. We recorded several operative parameters, such as volume of injected RTP, duration of occlusion and anastomotic time. Quality of occlusion was subjectively evaluated. Routine on-table angiography was performed to search for plug emboli. Primary patency, limb salvage and survival rates were reported at 6 months. RESULTS: In all patients, crural artery occlusion was achieved with the RTP without the use of an adjunct occlusion device. Mean volume of RTP used was 0.3 ml proximally and 0.25 ml distally. Mean duration of occlusion was 14.4 ± 4.5 min, while completion of the distal anastomosis lasted 13.4 ± 4.3 min. Quality of occlusion was judged as excellent in eight cases and good in 12 cases. Residual plugs were observed in two patients and removed with an embolectomy catheter, before we amended the technique for dissolution of RTP. At 6 months, primary patency rate was 75% but limb salvage rate was 87.5%. The 30-day mortality rate was 10%. CONCLUSIONS: This study shows that RTP is safe when properly dissolved and effective to occlude small calcified arteries for completion of distal anastomosis.

Information transfer between large and small two-dimensional polyacrylamide gel electrophoresis.

To determine the feasibility of data transfer, an interlaboratory comparison was conducted on colon carcinoma cell line (DLD-1) proteins resolved by two-dimensional polyacrylamide gel electrophoresis either on small (6 x 7 cm) or large (16x18 cm) gels. The gels were silver-stained and scanned by laser densitometry, and the image obtained was analyzed using Melanie software. The number of spots detected was 1337+/-161 vs. 2382+/-176 for small vs. large format gels, respectively. After gel calibration using landmarks determined using pl and Mr markers, large- and small-format gels were matched and 712+/-36 proteins were found on both types of gels. Having performed accurate gel matching it was possible to acquire additional information after accessing a 2-D PAGE reference database (http://www.expasy.ch/ cgibin/map2/def?DLD1_HUMAN). Thus, the difference in gel size is not an obstacle for data transfer. This will facilitate exchanges between laboratories or consultation concerning existing databases.

Optical characterization of sol-gel glasses derived from Eu3+ complex-forming precursors

Fabrication of new optical devices based upon the incorporation of rare earth ions via sol-gel methods depends on elimination of dopant ion clusters and residual hydroxyl groups from the final material. The optical absorption and/or luminescence properties of luminescent rare earth ions are influenced by the local bonding environment and the distribution of the rare-earth dopants in the matrix. Typically, dopants are incorporated into gel via dissolution of soluble species into the initial precursor sol. In this work, Eu3+ is used as optical probe, to assess changes in the local environment. Results of emission, excitation, fluorescence line narrowing and lifetimes studies of Eu3+-doped gels derived from Si(OCH3)4 and fluorinated/chelate Eu3+ precursors are presented. The precursors used in the sol-gel synthesis were: tris (6,6,7,7,8,8,8-heptafluoro-2,2-dimethyl-3,5-octanedionate) Eu(III), Eu (III) trifluoromethanesulfonate, Eu(III) acetylacetonate hydrate, Eu (III) trifluoroacetate trihidrate, tris (2,2,6,6-tetramethyl-3,5- heptanedionate) Eu(III) and Eu(NO3)3.6H2O. The results were interpreted in terms of the evolution of the Eu3+ fluorescence in systems varying from solution to the gels densified to 800ºC. The lifetimes studies indicate that the fluorinated precursors are effective at reducing the water content in densified gels.

Organo-gel de heptano: diagrama de fases e aplicações sintéticas

The phase diagram formation of microemulsion-based gels composed of an anionic surfactant aerosol-OT sodium bis (2-ethylhexyl)-sulphosuccinate), water, gelatin and an organic solvent is presented for heptane. The stability of this organo- gel, when an enzyme is immobilized is discussed in terms of its reutilization in various esters synthesis.

Influência da catálise ácida e básica na preparação da sílica funcionalizada pelo método sol-gel

Some aspects of the chemistry involved in the preparation and characterization of functionalized silicon oxide by sol-gel method are considered in this work. The synthesis was performed with different silicon alcoxide precursors and the influence of the acid and basic catalyst was investigated. Characterization was performed by infrared absorption spectroscopy, elemental analysis and 29Si NMR. Infrared data show Si-C and -CH2- vibrational modes at 1250 to 1280 and 2920 to 2940 cm-1, respectively. The elemental analysis confirmed the presence of organic groups in the inorganic silica network. 29Si NMR results show different hydrolisys depending on the acid or base catalysis.

Coexistence of Two Kinds of Fluorinated Hydrogenated Micelles as Building Blocks for the Design of Bimodal Mesoporous Silica with Two Ordered Mesopore Networks

A simple and effective route has been developed for the synthesis of bimodal (3.6 and 9.4 nm) mesoporous silica materials that have two ordered interconnected pore networks. Mesostructures have been prepared through the self assembly mechanism by using a mixture of polyoxyethylene fluoroalkyl ether and triblock copolymer as building block. The investigation of the RF8(EO)9/P123/water phase diagram evidences that in the considered surfactant range of concentrations, the system is micellar (L1). DLS measurements indicate that this micellar phase is composed of two types of micelles, the size of the first one at around 7.6 nm corresponds unambiguously to the pure fluorinated micelles. The second type of micelles at higher diameter consists of fluorinated micelles which have accommodated a weak fraction of P123 molecules. Thus, in this study the bimodal mesoporous silica are really templated by two kinds of micelles.

Ultra-trace monitoring of copper in environmental and biological samples by inductively coupled plasma atomic emission spectrometry after separation and preconcentration by using octadecyl silica membrane disks modified by a new schiff's base

Ultra-trace amounts of Cu(II) were separated and preconcentrated by solid phase extraction on octadecyl-bonded silica membrane disks modified with a new Schiff,s base (Bis- (2-Hydroxyacetophenone) -2,2-dimethyl-1,3-propanediimine) (SBTD) followed by elution and inductively coupled plasma atomic emission spectrometric detection. The method was applied as a separation and detection method for copper(II) in environmental and biological samples. Extraction efficiency and the influence of sample matrix, flow rate, pH, and type and minimum amount of stripping acid were investigated. The concentration factor and detection limit of the proposed method are 500 and 12.5 pg mL-1, respectively.

A novel method for fast enrichment and monitoring of hexavalent and trivalent chromium at the ppt level with modified silica MCM-41 and its determination by inductively coupled plasma optical emission spectrometry

Chromium(III) at the ng L-1 level was extracted using partially silylated MCM-41 modified by a tetraazamacrocyclic compound (TAMC) and determined by inductively coupled plasma optical emision spectrometry (ICP OES). The extraction time and efficiency, pH and flow rate, type and minimum amount of stripping acid, and break- through volume were investigated. The method's enrichment factor and detection limit are 300 and 45.5 pg mL-1, respectively. The maximum capacity of the 10 mg of modified silylated MCM-41 was found to be 400.5±4.7 µg for Cr(III). The method was applied to the determination of Cr(III) and Cr(VI) in the wastewater of the chromium electroplating industry and in environmental and biological samples (black tea, hot and black pepper).

Estudio clínico del tratamiento de las exóstosis digitales mediante la implantación de un gel biocompatible

Antecedentes: La exóstosis digital es una patología común que se presenta con frecuencia en las consultas de podología. A pesar del alto grado de incidencia es una entidad clínica muy poco estudiada. En la actualidad, los tratamientos propuestos para resolver esta patología son el tratamiento conservador mediante ortesis de silicona y el tratamiento quirúrgico consistente en la resección de la exostosis. Consideramos necesario otra alternativa de tratamiento como la infiltración de un gel de Poliacrilamida (PAAG) que presente mayores beneficios y menos inconvenientes a los pacientes que los tratamientos propuestos hasta el momento. Material y Métodos: Estudio realizado durante los meses de octubre de 2006 y febrero de 2009, sobre 54 pacientes (2 varones y 52 mujeres) de edades comprendidas entre 43 y 85 años afectados de exostosis digital. Se procede a la infiltración de un gel biocompatible en la zona de la exostosis para aislar la misma y evitar presiones según técnica de infiltración descrita. Todos los pacientes firman el documento de consentimiento informado. Los datos fueron recogidos de forma objetiva y subjetivamente en cada uno de los pacientes. La posterior evaluación de los datos y su tratamiento estadístico se llevó a cabo de forma disociada y anónima. El análisis estadístico se realizó con el paquete estadístico SPSS-PC 17.0. Resultados: El heloma interdigital en 4º espacio y 5º dedo se presentó en el 35,2% de los casos; el heloma en dorso del 5º dedo en el 29,6%; el heloma en dorso de dedo central en el 11,1% de los casos; heloma interdigital en 4º espacio y 4º dedo en el 9,3%; y el resto de lesiones primarias en un 14,9% del total de los casos. De cada uno de los casos se observó: tratamientos previos, cantidad de fármaco infiltrada, reacción al fármaco, controles y satisfacción del paciente al tratamiento. A un 57,4% de los pacientes no se les había realizado ningún tratamiento previo, mientras que a un 33,3% se le había realizado una ortesis de silicona. Analizando la cantidad de PAAG infiltrado en cada caso nos resultó una media de 0,542 cc con una desviación típica de 0,2237 cc. La cantidad mínima fue de 0,20 cc, hasta un máximo de 1,50 cc. La reacción observada al fármaco a las 24 y 48 horas de la infiltración fue prácticamente la misma que la del periodo inicial, observándose el habón en ambos tiempos de valoración. Se practicaron cuatro controles posinfiltración a la semana, al mes, a los tres meses y a los seis meses, en los que a seis meses pudimos concluir que en la mayoría de los casos ? 45 de 54 - persistió el habón de PAAG del inicio. La satisfacción se valora a la semana, al mes, a los tres meses y a los seis meses siendo a los 6 meses el grado de satisfacción de los pacientes del 98%. Conclusiones: El tratamiento del heloma provocado por exostosis digital mediante infiltración de gel de poliacrilamida según la técnica protocolizada en este estudio clínico es factible, reproductible y eficaz. Alcanzando el producto un alto nivel de tolerancia y adaptación a las estructuras anatómicas y disminuyendo los efectos secundarios, lo que nos hace afirmar que es una técnica segura con la que hemos obtenido óptimos resultados en lo que se refiere a efectividad del tratamiento y disminución del dolor. Podemos concluir que la infiltración de gel de poliacrilamida se convierte una nueva opción de tratamiento en el caso de exostosis digitales.

Estudio clínico del tratamiento de las exóstosis digitales mediante la implantación de un gel biocompatible

Antecedentes: La exóstosis digital es una patología común que se presenta con frecuencia en las consultas de podología. A pesar del alto grado de incidencia es una entidad clínica muy poco estudiada. En la actualidad, los tratamientos propuestos para resolver esta patología son el tratamiento conservador mediante ortesis de silicona y el tratamiento quirúrgico consistente en la resección de la exostosis. Consideramos necesario otra alternativa de tratamiento como la infiltración de un gel de Poliacrilamida (PAAG) que presente mayores beneficios y menos inconvenientes a los pacientes que los tratamientos propuestos hasta el momento. Material y Métodos: Estudio realizado durante los meses de octubre de 2006 y febrero de 2009, sobre 54 pacientes (2 varones y 52 mujeres) de edades comprendidas entre 43 y 85 años afectados de exostosis digital. Se procede a la infiltración de un gel biocompatible en la zona de la exostosis para aislar la misma y evitar presiones según técnica de infiltración descrita. Todos los pacientes firman el documento de consentimiento informado. Los datos fueron recogidos de forma objetiva y subjetivamente en cada uno de los pacientes. La posterior evaluación de los datos y su tratamiento estadístico se llevó a cabo de forma disociada y anónima. El análisis estadístico se realizó con el paquete estadístico SPSS-PC 17.0. Resultados: El heloma interdigital en 4º espacio y 5º dedo se presentó en el 35,2% de los casos; el heloma en dorso del 5º dedo en el 29,6%; el heloma en dorso de dedo central en el 11,1% de los casos; heloma interdigital en 4º espacio y 4º dedo en el 9,3%; y el resto de lesiones primarias en un 14,9% del total de los casos. De cada uno de los casos se observó: tratamientos previos, cantidad de fármaco infiltrada, reacción al fármaco, controles y satisfacción del paciente al tratamiento. A un 57,4% de los pacientes no se les había realizado ningún tratamiento previo, mientras que a un 33,3% se le había realizado una ortesis de silicona. Analizando la cantidad de PAAG infiltrado en cada caso nos resultó una media de 0,542 cc con una desviación típica de 0,2237 cc. La cantidad mínima fue de 0,20 cc, hasta un máximo de 1,50 cc. La reacción observada al fármaco a las 24 y 48 horas de la infiltración fue prácticamente la misma que la del periodo inicial, observándose el habón en ambos tiempos de valoración. Se practicaron cuatro controles posinfiltración a la semana, al mes, a los tres meses y a los seis meses, en los que a seis meses pudimos concluir que en la mayoría de los casos ? 45 de 54 - persistió el habón de PAAG del inicio. La satisfacción se valora a la semana, al mes, a los tres meses y a los seis meses siendo a los 6 meses el grado de satisfacción de los pacientes del 98%. Conclusiones: El tratamiento del heloma provocado por exostosis digital mediante infiltración de gel de poliacrilamida según la técnica protocolizada en este estudio clínico es factible, reproductible y eficaz. Alcanzando el producto un alto nivel de tolerancia y adaptación a las estructuras anatómicas y disminuyendo los efectos secundarios, lo que nos hace afirmar que es una técnica segura con la que hemos obtenido óptimos resultados en lo que se refiere a efectividad del tratamiento y disminución del dolor. Podemos concluir que la infiltración de gel de poliacrilamida se convierte una nueva opción de tratamiento en el caso de exostosis digitales.

Optimization of the carrot leaf dehydration aiming at the preservation of omega-3 fatty acids

The carrot leaf dehydration conditions in air circulation oven were optimized through response surface methodology (RSM) for minimizing the degradation of polyunsaturated fatty acids, particularly alpha-linolenic (LNA, 18:3n-3). The optimized leaf drying time and temperature were 43 h and 70 ºC, respectively. The fatty acids (FA) were investigated using gas chromatography equipped with a flame ionization detector and fused silica capillary column; FA were identified with standards and based on equivalent-chain-length. LNA and other FA were quantified against C21:0 internal standard. After dehydration, the amount of LNA, quantified in mg/100 g dry matter of dehydrated carrot leaves, were 984 mg.