Oligomerization of 10,16-dihydroxyhexadecanoic acid and methyl 10,16-dihydroxyhexadecanoate catalyzed by lipases.

The main monomer of tomato cuticle, 10,16-dihydroxyhexadecanoic acid (10,16-DHPA) and its methyl ester derivative (methyl-10,16-dihydroxyhexadecanote; methyl-10,16-DHHD), were used to study their oligomerization reactions catalyzed by five lipases: Candida antarctica lipase B (CAL-B), Rhizomucor miehei lipase (RM), Thermomyces lanuginosus lipase (TL), Pseudomonas cepacia lipase (PCL) and porcine pancreatic lipase (PPL). For 10,16-DHPA, optimum yields were obtained at 60 °C using toluene and 2-methyl-2-butanol (2M2B) as solvent, while for methyl-10,16-DHHD the bests yields were obtained in toluene and acetonitrile. Both reactions leaded to linear polyesters according to the NMR and FT-IR analysis, and there was no data indicating the presence of branched polymers. Using optimized conditions, poly(10,16-DHPA) and poly(methyl-10,16-DHHD) with Mw = 814 and Mn = 1,206 Da, and Mw = 982 and Mn = 860 Da, respectively, were formed according to their MALDI-TOF MS and ESI-MS data. The self-assembly of the polyesters obtained were analyzed by AFM.

Characterization and quantification of saponins and flavonoids in sprouts, seed coats and cotyledons of germinated black beans.

Saponins, flavonols and isoflavones were quantified in sprouts, cotyledons and seed coats of black beans (Phaseolus vulgaris L.) subjected to germination over five days. Sprouts had a higher concentration of saponins compared to cotyledons or seed coats (p<0.05). The saponins concentration in hilum increased 2.3-fold after soaking. After the first day of germination, the saponin concentration in sprouts and cotyledons increased 1.9 and 2.1-fold, respectively. Additional germination days decreased the amount of the most abundant soyasaponins in black bean sprouts. Flavonols and isoflavones were associated with seed coats and less than one third of the initial amount remained after the soaking process. The concentrations of flavonols were also reduced during germination process. Aglycones were detected only after soaking and their concentration remained unchanged during germination. Genistein was detected only after three days of germination. In general, one-day germinated black beans could be recommended for increasing the concentration of saponins and non-glycosylated flavonols in sprouts and seed coats, respectively.

Características cinemáticas, cinéticas e eletromiográficas do andar de adultos jovens com e sem suporte parcial de peso corporal / Kinematic, kinetic and electromyographic characteristics of young adults walking with and without partial body weight support

OBJETIVO: O objetivo deste estudo foi analisar as características cinemáticas, cinéticas e eletromiográficas do andar de adultos jovens em piso fixo sem colete e com suporte parcial de peso (SPP) de 0, 10, 20 e 30 por cento do peso corporal. MÉTODO: Oito jovens com idade média de 22,2 anos foram filmados andando sobre uma passarela que continha uma plataforma de força na região central para registro das componentes da força de reação do solo. Marcadores refletivos foram posicionados nos principais pontos anatômicos dos membros inferiores para registro dos dados cinemáticos, e eletrodos de superfície foram afixados nos músculos tibial anterior e gastrocnêmio medial para registro da atividade elétrica muscular. RESULTADOS: Diferenças significantes entre as cinco condições experimentais foram constatadas nas variáveis espaço-temporal, nos ângulos máximos e mínimos da coxa, joelho e tornozelo e nas amplitudes das componentes horizontal ântero-posterior e vertical da força de reação do solo. De forma geral, as maiores mudanças ocorreram na condição de SPP de 30 por cento do peso corporal. CONCLUSÃO: É importante considerar as compensações que ocorrem no padrão do andar com SPP no planejamento das intervenções terapêuticas. Ainda, para melhor definir a utilização dos sistemas de suspensão de peso na reabilitação, estudos futuros precisam ser realizados para verificar o comportamento do andar em populações com alteração de movimento em piso fixo.

OBJECTIVE: The aim of this study was to analyze the kinematic, kinetic and electromyographic characteristics of young adults walking on a fixed platform without a vest and with partial body weight support (PBWS) of 0, 10, 20 and 30 percent. METHOD: Eight young adults (mean age: 22.2 years) were videotaped walking on a walkway that contained a force plate embedded in its middle portion, to record the ground reaction force (GRF) components. Reflective markers were placed on the main anatomical points of the lower limbs in order to acquire kinematic data, and surface electrodes were attached to the tibialis anterior and gastrocnemius medialis muscles in order to record electromyographic muscle activity. RESULTS: Significant differences among the five experimental conditions were observed with regard to spatial-temporal variables, the maximum and minimum angles for the thigh, knee, and ankle, and the amplitudes of the anteroposterior horizontal and vertical GRF components. Generally, the greatest changes occurred with PBWS of 30 percent. CONCLUSION: It is important to take into consideration the compensations to walking patterns that occur with PBWS, in planning therapeutic interventions. Moreover, to better define the use of suspended weight systems in rehabilitation programs, further investigations should be conducted in order to verify the walking patterns on fixed platforms among populations with movement disorders.

Ectopic eruption and severe root resorption.

The treatment of a patient with maxillary canine-central incisor transposition is presented. Treatment options were limited because the roots of the maxillary right lateral and central incisors had severe resorption and generalized shortening. The multidisciplinary approach and treatment sequence used to achieve an esthetic and functional result are discussed in this case report.

La inhibición de la expresión de los antígenos clase I y calse II del Complejo Mayor de Histocompatibilidad en macrófagos infectados con Myrcobacterium tuberculosis H37Rv se debe a alteraciones en la vía JAK-STAT / Inhibition of class I and class II antigens of Major Histocompatibility Complex expression in Mycobactrium tuberculosis H37RV infected macrophages is caused by JAK-STAT way alterations

Objetivo: establecer los mecanismos moleculares por los cuales la infección de macrófagos murinos con Mycobacterium tuberculosis (Mtb), inhiben la expresión de los antígenos del complejo mayor de histocompatibilidad (MHC) inducidos por Interferón gamma (IFNγ). Diseño: experimental. Materiales y Métodos: la línea celular de macrófagos murinos B10R se infectó con Mtb cepa H37Rv, o se trató con micobacterias muertas por calor, paredes celulares de M. bovis y lipoarabinomanan manosilado (ManLAM) Mediciones: mediante citometría de flujo y Northern-Blot se evaluó la expresión superficial y los niveles de mRNA para los antigenos del MHC. La expresión superficial de las cadenas α y β del receptor de IFNγ (IFNγR), se evaluó por citometría de flujo. La evaluación de los niveles de fosforilación de JAK2 y STAT1 se realizó por inmunoprecipitación e inmunodeteccción. Para la estimación de los niveles del mRNA para CIITA se utilizo la técnica de RT-PCR. Resultados: La infeccióncon Mtb resultó en una inhibición de la expresión superficial de los antígenos clase I y II, y de los niveles de mRNA para clase II. Micobacterias muertas por calor y paredes celulares micobacterianas también causaron esta inhibición, pero no el tratamiento con ManLAM. La infección también causó una disminución en la fosforilación de JAK2 y STAT1, pero no una disminución en la expresión superficial del IFNγR. Adicionalmente, los niveles de mRNA para CIITA también fueron inhibidos. Conclusión: La capacidad de la Mtb para evadir la respuesta inmune puede deberse a su habilidad para alterar la respuesta al IFNγ, y la supresión de las moléculas clase II

[Influence of the pericardium in the pathophysiology of ventricular dysfunction in acute infarct of the right ventricle. Experimental study] . / La influencia del pericardio en la patofisiología de la disfunción ventricular en el infarto agudo del ventrículo derecho. Estudio experimental.

UNLABELLED: To obtain more information about the role of the pericardium in the setting of acute right ventricular infarction (ARVI) we studied the behaviour of the ventricular function curves (VFC) and the relationship of the ventricular end-diastolic pressures (R-VEDP-RV:LV) in two groups of dogs. Group A. (n = 12) Control (C), ARVI, Pericardiectomy (P). A parabolic behaviour of the C VFC was noted (r2 = 071) and it's flexion point (FP) was found in 13. +/- 2 mmHg. After the ARVI the right (R) VFC was shifted downwards and to the right and the FP was documented in 18 +/- 2 mmHg (p < 0.05) in relation to C VFC. After P the RVFC was displaced upwards and to the left in relation to ARVIC RVFC (p < 0.05). The C R-VEDP-RV:LV = 0.75 and only a trend to equalization after the ARVI and after P were noted (0.91, 0.84, respectively) (p = ns). Group B (n = 12). Control (C), P, ARVI. The RVFC after P was shifted up and to the left in relation to the C RVFC (p < 0.05) and the FP = 10 +/- 2 mmHg. After P in the setting of ARVI the RVFC was shifted downward and to the right in relation to P RVFC (p < 0.05). After P the R-VEDP-RV:LV = 0.45 and statistical significant equalized in the condition of ARVI without pericardium (0.95, p < 0.05). CONCLUSION: Ours results support a partial restrictive role of the pericardium in the origin of the low cardiac output (LCO) in ARVI. Because, equalization of the R-VEDP-RV:LV is not only due to the restraining pericardial effect but is also due to right ventricular myocardial ischemia. The FP (18. +/- 2 mmHg) found seems to be the top value of RVEDP for volume infusion in experimental ARVI. Hemodynamic finding that could be useful in the preload volume management for humans with ARVI and LCO or systemic hypotension.

[Ventricular function curves with and without pericardium: analysis of pericardial pressures]. / Las curvas de función ventricular con y sin pericardio: análisis de las presiones pericárdicas.

Simultaneous right and left ventricular function curves (VFC, R, L) were obtained in a canine model, (open chest preparation), with and without pericardium. Preload and afterload conditions for the right and left ventricles were controlled. VFC were constructed from zero to 25 mmHg of ventricular end-diastolic pressures and by increasing the cardiac output from 50 to 250 mL/kg-1min-1. Both, right and left VFC showed an initial steep rise at low filling pressures and then flattened off to a plateau at high filling pressures. The best mathematical model that fitted with the VFC, with and without the pericardium was the parabola (r2 = 0.71, 0.72 respectively). After pericardiectomy R and L VFC were displaced to the left of the VFC with pericardium and a decrease in filling pressures were noted at the same points of cardiac output, findings that suggest a restraining effect of the pericardium. By subtracting the filling pressures obtained with pericardium from those without pericardium at the same levels of cardiac output, pericardial pressures were derived. In all the range of the VFC the pericardial pressures were positive, and this pressure increase as cardiac output increase. Thus the transmural pressure was never cero, for both right and left ventricles. The observed relation for the R and L filling pressures, derived from a polynomial equation of second order suggest a small although not unimportant effect of the pericardium at normal filling pressures, and a very substantial influence at high levels of cardiac output. The demarcation between small and major effects appears in the upper range of normal filling pressures in this dynamic approach of the pericardial pressures.

Resistencia a los antibioticos de patógenos bacterianos aislados de infecciones sistémicas : estudio cooperativo / Resistance to antibiotics of bacterian pathogens isolated from sistemic infections

La emergencia bacteriana a diferentes antibióticos constituye un problema alarmante. En la situación actual, para un adecuado tratamiento de pacientes con infecciones severas, es vital la correcta identificación del patógeno y el estudio de su patrón de sensibilidad. Con ese objetivo, se ralizó una experiencia piloto en la que participaron los laboratorios de bacteriología clínica de tres hospitales-centinela, en colaboración con el Departamento de laboratorios de Salud Pública (DLSP). Los laboratorios de cada hospital registraron todas las cepas invasivas aisladas de pacientes internados entre octubre de 1996 y abril de 1997, y las refirieron al DLSP conjuntamente con datos básicos del paciente y de la bacteria. En el DLSP se confirmó o completó la identificación y la susceptibilidad de los agentes. Se estudiaron 299 cepas invasivas, pertenecientes a 29 géneros/especies diferentes, de las cuales 54 por ciento provenían de infeccions intrahospitalarias. Los bacilos Gram negativos predominaron: E. coli (29); Acinetobacter sp. (25), K. pneumoniae (21), E. cloacae (11), S. marcescens (11), P aeruginosa (10) y K.oxytoca (6). Prácticamente todos los patrones de resistencia descritos en la literatura para estas especies, fueron registrados en esos aislamientos. El monitoreo efectuado demostró la factibilidad de coordinar acciones de vigilancia de la resistencia a antibióticos, así como la conveniencia de lograr conjuntos de datos referidos al tema. Permitió confirmar la disminución de la susceptibilidad de N. meningitidis a la penicilina (50 por ciento), y ratificar la preeminencia intrahospitalaria de las bacterias Gram positivas. S aureus fue frecuente (n=67), con 27 por ciento de cepas resistentes a la meticilina y la mayoría de éstas sólo sensibles a vancomicina. S. pneumoniae (n=50) fue el patógeno comunitario dominante, con 26 por ciento de resistencia a la penicilina, especialmente en cepas de niños. No se detectaron Enterococcus ni S. aureus resistentes a vancomicina. El riesgo de la aparición de esa resistencia, sumado a la progresión de la resistencia de diferentes agentes a diversos fármacos, constituye un elemento decisivo para implementar y mantener un sistema nacional de monitoreo que permita registrar tendencias de los patrones de sensibilidad y que alerte precozmente respecto a cambios drásticos en el espectro de susceptibilidad en diferentes especies bacterianas

Cloning, expression and toxicity of a mosquitocidal toxin gene of Bacillus thuringiensis subsp. medellin.

Bacillus thuringiensis (Bt) subsp. medellin (Btmed) produces parasporal crystalline inclusions which are toxic to mosquito larvae. It has been shown that the inclusions of this bacterium contain mainly proteins of 94, 68 and 28-30 kDa. EcoRI partially digested total DNA of Btmed was cloned by using the Lambda Zap II cloning kit. Recombinant plaques were screened with a mouse polyclonal antibody raised against the 94 kDa crystal protein of Btmed. One of the positive plaques was selected, and by in vivo excision, a recombinant pBluescript SK(-) was obtained. The gene encoding the 94 kDa toxin of Btmed DNA was cloned in a 4.4 kb DNA fragment. Btmed DNA was then subcloned as a EcoRI/EcoRI fragment into the shuttle vector pBU4 producing the recombinant plasmid pBTM3 and used to transform by electroporation Bt subsp. israelensis (Bti) crystal negative strain 4Q2-81. Toxicity to mosquito larvae was estimated by using first instar laboratory reared Aedes aegypti, and Culex quinquefasciatus larvae challenged with whole crystals. Toxicity results indicate that the purified inclusions from the recombinant Bti strain were toxic to all mosquito species tested, although the toxicity was not as high as the one produced by the crystal of the Btmed wild type strain. Polyacrylamide gel electrophoresis indicate that the inclusions produced by the recombinant strain Bti (pBTM3) were mainly composed of the 94 kDa protein of Btmed, as it was determined by Western blot.

[Assessment of genetic toxicity of the exposure to clomiphene citrate, with various bacterial test systems]. / Valoración de riesgo gentóxico de exposición a citrato de clomifeno, con diferentes sistemas de prueba bacterianos.

Clomiphene citrate (CC) induces framshift mutations on the Salmonella typhimurium Ames strains TA1538, TA97 and TA100 employing in vitro metabolic activation with S9 aerochlor 1254 induced rat livers, but not base pair substitution mutations with neither the standard or the preincubation method. CC induced genolethal DNA damages on the Escherichia coli PolA-/PolA+ with S9 on the preincubation method or without S9 on the disk diffusion one. The severe primary DNA damages produced b CC was verified by the SOS induction on the lysogenic lambda phage induction with De Marini (1988) method and the induction of colicin E1 plasmid on E. coli. These results are suggestive that CC may be an adduct forming compound which is able to inhibit replication if the cell lacks DNA polymerase, or it may produce framshift mutations after replications. CC induced damages could be large lesions conducing to unicatenary DNA strains, that are able to induce the lexA regulated genes. So, the use of this ovulation inductor is a risk of genotoxic damage and it is advisable to do a risk-benefit evaluation in any particular case before its prescription.

Biochemical, immunological and toxicological characteristics of the crystal proteins of Bacillus thuringiensis subsp. medellin.

Characterization of the insecticidal and hemolytic activity of solubilized crystal proteins of Bacillus thuringiensis (Bt) subsp. medellin (Btmed) was performed and compared to solubilized crystal proteins of isolates 1884 of B. thuringiensis subsp. israelensis (Bti) and isolate PG-14 of B. thuringiensis subsp. morrisoni (Btm). In general, at acid pH values solubilization of the Bt crystalline parasporal inclusions (CPI) was lower than at alkaline pH. The larvicidal activity demonstrated by the CPI of Btmed indicated that optimal solubilization of CPI takes place at a pH value of 11.3, in Bti at pH values from 5.03 to 11.3 and in Btm at pH values from 9.05 to 11.3 Hemolytic activity against sheep red blood cells was mainly found following extraction at pH 11.3 in all Bt strains tested. Polyacrylamide gel electrophoresis under denaturing conditions revealed that optimal solubilization of the CPI in all Bt strains takes place at the alkaline pH values from 9.05 to 11.3. An enriched preparation of Btmed crystals was obtained, solubilized and crystal proteins were separated on a size exclusion column (Sephacryl S-200). Three main protein peaks were observed on the chromatogram. The first peak had two main proteins that migrate between 90 to 100 kDa. These proteins are apparently not common to other Bt strains isolated to date. The second and third peaks obtained from the size exclusion column yielded polypeptides of 68 and 28-30 kDa, respectively. Each peak independently, showed toxicity against 1st instar Culex quinquefasciatus larvae. Interestingly, combinations of the fractions corresponding to the 68 and 30 kDa protein showed an increased toxicity. These results suggest that the 94 kDa protein is an important component of the Btmed toxins with the highest potency to kill mosquito larvae. When crystal proteins of Bti were probed with antisera raised independently against the three main protein fractions of Btmed, the only crystal protein that showed cross reaction was the 28 kDa protein. These data suggest that Btmed could be an alternative bacterium for mosquito control programs in case mosquito larval resistance emerges to Bti toxic proteins.

Transfer of toxin genes to alternate bacterial hosts for mosquito control.

Mosquitoes are vector of serious human and animal diseases, such as malaria, dengue, yellow fever, among others. The use of biological control agents has provide an environmentally safe and highly specific alternative to the use of chemical insecticides in the control of vector borne diseases. Bacillus thuringiensis and B. sphaericus produce toxic proteins to mosquito larvae. Great progress has been made on the biochemical and molecular characterization of such proteins and the genes encoding them. Nevertheless, the low residuality of these biological insecticides is one of the major drawbacks. This article present some interesting aspects of the mosquito larvae feeding habits and review the attempts that have been made to genetically engineer microorganisms that while are used by mosquito larvae as a food source should express the Bacillus toxin genes in order to improve the residuality and stability in the mosquito breeding ponds.

[Simple ureterocele diagnosed in utero. Presentation of a case]. / Ureterocele simple diagnosticado en útero. Presentación de un caso.

We report a case of simple ureterocele which was diagnosed in utero using prenatal ultrasonography at 32 weeks and subjected to serial observations in order to spot hydronephrosis and hydroureter. The diagnosis was confirmed at birth using ecosonography and the newborn was submitted to corrective surgery during the immediate neonatal period, obtaining excellent results.

A technique to collect and dislodge conidia produced by Paracoccidioides brasiliensis mycelial form.

Collection and separation of mycelial propagules produced by P. brasiliensis was accomplished by agitation with glass beads, centrifugation and filtration through cotton wool. The mean number of conidia liberated per plate (approximately 1 000 000) and their viability (79%), lead us to think that it is now possible to undertake experimental studies with these propagules.

Effect of temperature on the mycelium to yeast transformation of Paracoccidioides brasiliensis.

The influence of temperature on Paracoccidioides brasiliensis mycelium to yeast transformation was studied by sequential microscopic observations of slide cultures. When incubated at temperatures above 28 degrees C, the mycelial elements gradually produced round to oval chlamydospores and later on, exhibited multiple budding. A sizeable proportion of mycelial elements transformed at 34 degrees C; however, multiple budding was important only at 37 degrees C.