Microencapsulation of eugenol molecules by ß-cyclodextrine as a thermal protection method of antibacterial action.

Eugenol is natural oil that has excellent antibacterial properties but cannot be used to fabricate many products that require thermal processing. One possible alternative to the use of the eugenol molecules in high-temperature processes is the encapsulation of these molecules in a structure that is not toxic and is resistant to thermal treatment. This work investigated the encapsulation process of eugenol molecules in ß-cyclodextrine and the antibacterial properties of eugenol-ß-cyclodextrine (the eugenol-ßCD complex) against Escherichia coli and Staphylococcus aureus. The FTIR, DSC, MEV and TGA results show that the encapsulation method is an excellent alternative to increase the thermal stability of eugenol molecules. A value of 241.32L.mol-1 was determined for the formation constant (Kc) of the eugenol-ßCD complex, which confirmed the success of the encapsulation process. The MEV analysis shows the formation of approximately 12µm microcapsules. After the thermal treatment of the eugenol-ßCD complex at a temperature of 80°C for 2h, the complex retained significant antibacterial action, which confirms the thermal protection of the eugenol molecules. The minimum inhibitory concentration (MIC) and agar diffusion results show that the microcapsules containing 17.08mmol.L-1 of eugenol exhibited excellent antibacterial action against Escherichia coli and Staphylococcus aureus after thermal treatment.

Ultrastructural study of the denervated diaphragm in rats / Estudio ultraestructural del diafragma desnervado en ratas

The structural alterations that occur in the muscle fibers of denervated rat diaphragms were studied. Fifteen adult male albino rats (Rattus norvegicus) with a mean weight of 200 g and about 60 days of age were used. Chronically denervated diaphragms were obtained and the animals were sacrificed after 4, 8 and 12 weeks of denervation. The left antimere of the diaphragm was denervated by sectioning of the phrenic nerve and the right antimere served as control. Each antimere was divided into fragments, which were used for analysis transmission electron microscopy. During the initial phase of denervation (4 weeks), ultrastructural muscle modifications appeared in scattered fibers and in foci along these fibers. Muscle fibers with foci of less dense and loosely arranged myofibrils, disorganized Z line, displaced T tubules, and central nucleus exhibiting reentrances and fragmented aspect were observed. After 8 weeks, formation of large aggregates of small elongated mitochondria showing altered cristae, matrix inclusions and increased electron density was noted. At 12 weeks of denervation the alterations were found to be more drastic. Nuclei with internal deposits of myofibrillar or amorphous material were observed. In these fibers, vacuoles harbored enormous myeloid structures in the subsarcolemmal or intermyofibrillar region.

Fueron estudiadas las alteraciones ultra estructurales de las fibras musculares del diafragma denervado de ratas. Fueron utilizadas 15 ratas albinas (Rattus norvegicus), machos, adultas, con peso promedio de 200 g de aproximadamente 60 días de edad. Los diafragmas crónicamente denervados fueron obtenidos después de 4, 8 y 12 semanas de denervación. El antímero izquierdo del diafragma fue denervado por sección del nervio frénico y el antímero derecho fue utilizado como control. Cada antímero fue dividido en fragmentos, que fueron utilizados para el estudio en microscopia electrónica de transmisión. Durante los períodos iniciales de denervación (4 semanas), las modificaciones en la ultraestructura del músculo se disponen en fibras dispersas y en focos a lo largo de las mismas. Se observan fibras musculares con focos de miofibrillas rarefactas y laxamente dispuestas; línea Z desorganizada; túbulos T dislocados; núcleo central con aspecto fragmentado. Después de 8 semanas de denervación, se observa la formación de numeroso agregados de pequeñas mitocondrias alargadas, con alteraciones en las crestas, inclusiones en la matriz y aumento de la electrodensidad. Con 12 semanas de denervación, las alteraciones se muestran más drásticas; se observan núcleos con depósitos internos de material miofibrillar o amorfo. En estas fibras, las vacuolas presentan grandes estructuras mieloides en la región subsarcolemal o intermiofibrillar.

Morphologic alterations resulting from denervation of the diaphragm in rats / Alteraciones morfológicas resultantes de la denervación del diafragma en ratas

The morphological alterations that occur in the muscle fibers of denervated rat diaphragms were studied. Fifteen adult male albino rats (Rattus norvegicus) with a mean weight of 200 g and about 60 days of age were used. Chronically denervated diaphragms were obtained and the animals were sacrificed after 4, 8 and 12 weeks of denervation. The left antimere of the diaphragm was denervated by sectioning of the phrenic nerve and the right antimere served as control. Each antimere was divided into two fragments, which were used for histological (H.E.) and histoenzymological (NADH-TR and myofibrillar ATPase). After 4 weeks, denervated muscle fibers showed important light microscopic alterations: atrophy with angular profiles in cross-sections, cytoplasm containing vacuoles, enlarged interstitial space with increased connective tissue, cellular infiltration, and muscle fibers without defined contours. The most marked alterations being observed for type IIb and IIa fibers. Eight and 12 weeks after denervation, the NADH-TR reaction showed that it was impossible to characterize the muscle fibers based on their metabolic profile...

Fueron estudiadas las alteraciones morfológicas de las fibras musculares del diafragma denervado de ratas. Se utilizaron 15 ratas albinas (Rattus norvegicus) machos, adultos, con peso promedio de 200g y cerca de 60 días de edad. Se denervó el diafragma y después de 4, 8 y 12 semanas los animales fueron sacrificados. El antímero izquierdo del diafragma fue denervado por sección del nervio frénico y el antímero derecho fue utilizado como control. Cada antímero fue dividido en fragmentos, utilizados para el estudio histológico (H-E), histoenzimológico (NADH-TR y ATPasa miofibrilar). Después de 4 semanas las fibras musculares denervadas presentaron alteraciones importantes en lamicroscopía de luz: atrofia con perfiles angulados en secciones transversales; citoplasma con vacuolas; aumento del espacio intersticial con aumento de tejido conjuntivo; infiltración celular y fibras musculares sin contornos definidos, siendo las alteraciones más marcadas en las fibras tipo IIb e IIa. Después de 8 y 12 semanas de denervación la reacción para NADH-TR demuestra que es imposible caracterizar a las fibras musculares a través de su perfil metabólico...

Morphometric and morphological analysis of neuromuscular junction alterations in the denervated rat diaphragm / Evaluación de las alteraciones morfológicas y morfométricas de las uniones neuromusculares del diafragma denervado en ratas

The morphological and structural alterations that occur in the neuromuscular junctions of the denervated rat diaphragm were studied. Fifteen adult male albino rats (Rattus norvegicus) aged about 60 days and with a mean weight of 200 g were used. Chronically denervated diaphragms were obtained and the animals were sacrificed after 4, 8 and 12 weeks of denervation. The left antimere of the diaphragm was denervated by section of the phrenic nerve and the right antimere was used as control. Each antimere was divided into three fragments: one was used for histochemical (nonspecific esterase) and morphometric study of neuromuscular junctions, and the other two were used for transmission and scanning electron microscopy (SEM) analysis. Histochemical analysis of the diaphragm neuromuscular junctions after denervation showed only small changes in junction morphology. However, these junctions became smaller and elongated and presented less visible contours with increasing time of denervation. Ultrastructural analysis of neuromuscular junctions after 12 weeks showed more or less organized junctional folds on the muscle fiber surface. The junctional cytoplasm exhibited important alterations such as mitochondrial degeneration and the presence of numerous filaments. SEM revealed the presence of deep primary synaptic grooves with peripheral excavations which housed the nerve terminal boutons and exhibited internally the secondary synaptic clefts present among the junctional folds of the sarcolemma. This study showed that some of the morphological changes demonstrated in other denervated striated skeletal muscles are not repeated at the same intensity or in the same temporal pattern in the rat diaphragm.

En este trabajo se estudiaron las alteraciones morfológicas y estructurales de las uniones neuromusculares en el diafragma denervado de ratas. Se utilizaron 15 ratas albinas (Rattus norvegicus), machos, adultos, con peso promedio de 200g y cerca de 60 días de edad. Los diafragmas crónicamente denervados fueron obtenidos y los animales se sacrificaron después de 4, 8 y 12 semanas de denervación. El antímero izquierdo del diafragma fue denervado por sección del nervio frénico y el antímero derecho fue utilizado como control. Cada antímero fue dividido en 3 fragmentos: uno fue utilizado para el estudio histoquímico (esterasa inespecífica) y morfométrico. Los otros dos se destinaron al estudio de microscopía electrónica de transmisión (MET) y microscopia electrónica de barrido (MEB) de las uniones neuromusculares. El estudio histoquímico de las uniones neuromusculares posterior a la denervación, muestra que la morfología de esas uniones sufre pequeñas alteraciones. Con la evolución del tiempo de denervación esas uniones muestran tamaños menores, son alargadas y con contornos menos nítidos. La ultra-estructura de las uniones neuromusculares después de 12 semanas, demostró que la superficie de la fibra muscular exhibe pliegues de unión más o menos organizados. La región del citoplasma de unión exhibe alteraciones importantes, con degeneración mitocondrial y presencia de muchos filamentos. En MEB se observa que los botones sinápticos primarios son profundos, presentan escavaciones periféricas donde estaban alojados los botones de las terminaciones nerviosas y exhiben internamente, los espacios sinápticos secundarios presentes entre los pliegues de unión del sarcolema. Este estudio mostró que algunos patrones morfológicos demostrados en otros músculos estriados esqueléticos denervados no se repiten con la misma intensidad y curso temporal en el diafragma de ratas.

Histochemical and SEM evaluation of the neuromuscular junctions from alcoholic rats.

In the present study morphological changes occurring in the neuromuscular junctions (NMJ) of the extensor digitorum longus (EDL) and soleus muscles from albino rats (Rattus norvegicus) submitted to experimental chronic alcoholism were evaluated. Seventy two male animals aged 4 months and weighing on average 400g were divided into three groups: control, alcoholic and isocaloric. Six rats from each group were anesthetized and sacrificed after 5, 10, 15 and 18 months. The NMJ did not show detectable morphological changes in either muscle after treatment when examined by light microscopy. With respect to the dimensions, statistical analysis demonstrated a tendency to a statistically significant treatment x time interaction for the length of soleus muscle NMJ. The ultrastructural study, however, revealed that the NMJ of the soleus muscle of animals submitted to 18 months of experimental alcoholism presented important morphological alterations. Characteristically, the NMJ of these muscles is located on an elevation on the surface of the muscle fiber, presenting a regular round, oval or elliptical shape and continuous and not very deep synaptic grooves. Approximately 30% of the NMJ of alcoholic rats are irregular in shape, with the sarcolemmal elevations typical of the synapse region being flattened on at least one side, with discontinuous synaptic grooves, and deep and punctiform contacts of the synaptic buds. These data suggest that, although skeletal muscle has a greater natural resistance against the direct or indirect effects of alcohol, some submicroscopic morphological alterations are detectable in the NMJ, especially in muscles with oxidative metabolism (soleus) following long periods of ingestion of alcohol.

[AIDS, activism, and the regulation of clinical trials in Brazil: Protocol 028]. / AIDS, ativismo e regulação de ensaios clínicos no Brasil: o Protocolo 028.

This paper examines the politics and practices of drug evaluation in Brazil. It traces the history of AIDS activists' influence on the organization of modern clinical trials and their scientific rationale. Using the Merck indinavir trial as a case study, the authors discuss how organized civil society has developed strategies to intervene in the course of drug evaluation trials, shaping them according to its own interests. Adopting translation sociology as the theoretical framework, the paper describes and analyzes the strategies used by activists from "Grupo PelaVidda/SP" (an AIDS NGO) to build a consensus concerning indinavir monotherapy's lack of efficacy. The study considers the several regulatory forums involved in dealing with the controversy during the trial period.