Aspectos neuroendocrinos de la obesidad / Neuroendocrine aspects of obesity

En la fisiopatología de la obesidad intervienen factores genéticos, sociales, metabólicos, endocrinos y neurológicos. Esta multifactoriedad junto al hecho que estos factores se interrelacionan a través de mecanismos muy complejos, que son sólo parcialmente conocidos, ha llevado a que la comprensión íntima de este trastorno resulte una tarea sumamente ardua. Por estos motivos, el conocimiento integral de esta afección plantea un desafío al que actualmente están abocados numerosos grupos de investigadores. El análisis de la obesidad como un trastorno neuroendocrino, propone el estudio de este fenómeno desde una visión particular que implica disfunciones en casi todos los órganos endocrinos y en el sistema nervioso central, fundamentalmente en la actividad hipotalámica. Estas alteraciones afectan principalmente a los ejes neuroendocrinos hipotálamo-hipofiso-adrenal, adipo-insular y al control hipotalámico, tanto de la ingesta de alimento como del almacenamiento y gasto energético. Este artículo plantea una actualización en este campo; en primer lugar, se realiza una breve descripción, en forma independiente, de los principales sistemas antes mencionados y luego una descripción de su funcionamiento normal integrado. Finalmente, se describen desregulaciones de estos mecanismos y se discute como ellas contribuirían al desarrollo y/o mantenimiento de la obesidad.(AU)

Micropropagation of Ilex dumosa (Aquifoliaceae) from nodal segments in a tissue culture system

Micropropagation of Ilex dumosa var. dumosa R. ("yerba señorita") from nodal segments containing one axillary bud was investigated. Shoot regeneration from explants of six-year-old plants was readily achieved in 1/4 strength Murashige and Skoog medium (1/4 MS) plus 30 gr x L(-1) sucrose and supplemented with 4.4 microM BA. Further multiplication and elongation of the regenerated shoots were obtained by subculture in a fresh medium of similar composition with 1.5 gr x L(-1) sucrose. Rooting induction from shoots were achieved in two steps: 1) 7 days in 1/4 MS (30 gr x L(-1) sucrose, 0.25% Phytagel) with 7.3 microM IBA and 2) 21 days in the same medium without IBA and 20 microM of cadaverine added. Regenerated plants were successfully transferred to soil. This micropropagation schedule can be implemented in breeding programs of Ilex dumosa. (AU)

Distribution of pectins in the pollen apertures of Oenothera hookeri: velans ster/+ster

Cell wall pectins are some of the most complex biopolymers known, and yet their functions remain largely mysterious. The aim of this paper was to deepen the study of the spatial pattern of pectin distribution in the aperture of Oenothera hookeri.velans ster/+ster fertile pollen. We used "in situ" immunocytochemical techniques at electron microscopy, involving monoclonal antibodies JIM5 and JIM7 directed against pectin epitopes in fertile pollen grains of Oenothera hookeri.velans ster/+ster. The same region was also analyzed by classical cytochemistry for polysaccharide detection. Immunogold labelling at the JIM7 epitope showed only in mature pollen labelling mainly located at the intine endo-aperture region. Cytoplasmic structures near the plasma membrane of the vegetative cell showed no labelling gold grains. In the same pollen stge the labelling at the JIM5 epitope was mostly confined to a layer located in the limit between the endexine and the ektexine at the level of the border of the oncus. Some tubuli at the base of the ektexine showed also an accumulation of gold particles. No JIM5 label was demonstrated in the aperture chamber and either in any cytoplasmic structure of the pollen grains. The immunocytochemical technique, when compared with the traditional methods for non-cellulose polysaccharide cytochemistry is fare more sensitive and allows the univocal determination of temporal and spatial location of pectins recognized by the JIM7 and JIM5 MAbs. (AU)

Morphological changes in garlic (Allium sativum L. ) microbulblets during dormancy and sprouting as related to peroxidase activity and gibberellin A3 content

The aim of this work was to study the physiological mechanisms of dormancy and sprouting during post-harvest of garlic (Allium sativum L.) microbulblets produced by meristem culture of garlic seed cloves. The morphological changes occurring in garlic microbulblets were assessed from harvest till sprouting in relation with peroxidase activity and levels of gibberellins. Also the effect of a cold treatment (30 days at 4 degrees C) given 30 days after harvest was studied. The results showed that during the state of dormancy in garlic microbulblets formation of the leaf primordia and vascular differentiation of the storage leaf occurred, while increases of peroxidase activity and low levels of GA3 (the only active gibberellin identified) were found. At the end of dormancy the sprouting channel was formed, vascular differentiation established, and peaks of soluble peroxidase activity as well as of GA3 were observed. At day 90 post-harvest, garlic microbulblets showed physiologically mature and able to sprout. Further on, bud expansion and decrease of GA3 levels characterized sprouting of the microbulblets. The cold treatment enhanced GA3 levels and anticipated the sprouting process.(AU)

Morphogenesis in short-term and long-term anther derived calli of Oenothera hookeri de vries

Anther culture of O. hookeri on Murashige and Skoog (1962) medium supplemented with 2 mg l-1 2,4-dichlorophenoxyacetic acid and 2 mg-1 1-naphthaleneacetic acid produced callus formation. When subcultured onto medium lacking auxin, the callus regenerated through the organogenic pathway. Non-organogenic and organogenic callus was observed using histological methods after 2, 3 and 24 weeks in culture. Three types of calli were recognized: non-organogenic friable calli, organogenic friable calli with roots and organogenic hard calli with shoots. The microscopical sections showed striking differences in tissue organization among friable and compact calli. Vascular bundles were prominent in compact calli, but were not found in friable calli. Calli sections showed at light microscopy cells at two developmental stages; differentiated highly vacuolated cells and meristematic small isodiametric cells with densely stained cytoplasm. At electron microscopy level abnormal chloroplasts were present in non-organogenic calli, while chloroplasts were well developed in organogenic hard calli. Peroxisomes with paracrystalline protein bodies were abundant in both types of calli.(AU)

Plant regeneration from single-nodal-stem explants of legume tree Prosopis alba Griseb

Seeds of Prosopis alba were scarified with abrasive paper and placed to germinate on MS (Murashige and Skoog 1962) nutrient medium. After 7 days of culture, the basal part of cotyledons was removed and pieces of 4 mm" from distal parts were cultured on Murashige and Skoog (1962) mineral salts and vitamins (MS) (3 sucrose) supplemented with growth regulators. Callus proliferation took place in the majority of the media tested. A low percentage of calluses with green buds that developed on MS basal medium containing 0.1 mg.L-1 2,4-D alone or supplemented with BAP at 0.1 mg.L-1 was observed. Neither cotyledonary segments in any medium assayed regenerated the whole plants. Bud elongation (near 70) was achieved when single-nodal-stem segments cut from 20 days old seedlings were cultured on MS salts supplemented with 3 mg.L-1 NAA or 3 mg.L-1 IBA combined with 0.05 mg.L-1 KIN after 60 days in culture. Multiple shoots per bud were also observed. Single-nodal-stem segments from five-year-old plants were also cultured on the same media used for seedling explants. Maximal frequency of explants with bud elongation (near 70) was found on MS with 0.1 mg.L-1 NAA plus 1 mg.L-1 BAP after 60 days of culture. Single-nodal-stem explants cut from adult trees (more than 20 years) were also employed, but the number of bud elongation was lesser. For rooting, the elongated shoots were transferred to a semisolid or liquid MS culture medium employing a paper bridge, supplemented with 0.5 mg.L-1 IBA or 0.1 mg.L-1 NAA.(AU)

Factors influencing protease production by two Antarctic strains of Stenotrophomonas maltophilia

The influence of culture medium buffer capacity, the supplementation of culture medium with L-ala and the requirement of calcium for exoprotease production by Antarctic psychrotrophic Stenotrophomonas maltophilia strains ANT-1-1 and ANT-7-1 were examined. When increasing concentrations of calcium chloride (0 to 0.3 g l-1) were added to culture media, maximum protease production yields increased 70-75 (ANT-1-1) and 50 (ANT-7-1), while biomass levels showed little difference. Calcium was also necessary for optimal activity of proteases. L-ala had no effect on protease production. The reduction in buffer capacity, with the consequent change in external pH, had a positive effect, enhancing protease yields. Secretion of proteases into the medium started at the beginning of the stationary phase, corresponding with a rise in pH values up to pH 8.7 and was maximal at 36 h of culture. These results indicate that the regulation of calcium concentration and buffer capacity and also pH monitoring are factors to be considered when the design of an industrial culture medium and the optimisation of protease production processes using these Antarctic strains are concerned.(AU)

In vitro propagation of Berberis buxifolia Lam

Berberis buxifolia is a native shrub of Patagonia with a great importance due to its crop production as soon its medicinal and tinctorial applications. The aim of this work was to develop a protocol for in vitro propagation of B. buxifolia, with special emphasys on the rooting stage. The culture of the explants on Murashige and Skoog (1962) medium added with 0.55 microM BA allowed to attain a multiplication rate of 1:4.7 at day 63. Rooted shoots were obtained on Murashige and Skoog medium with half strength of macronutrient salts. The culture of the shoots with a period of 4 days of darkness at the beginning of the rooting, on a medium with 1.25 microM IBA for 7 days, followed by a IBA free medium until day 28, allowed to attain 80 rooting. These results show that B. buxifolia can be in vitro propagated.(AU)

Plant regeneration of Ilex paraguariensis (Aquifoliaceae) by in vitro culture of nodal segments

In vitro regeneration of complete plants from nodal single bud segments of "yerba mate" (Ilex paraguariensis St. Hil.) was studied under defined nutritional and environmental conditions. Nodal segments harvested from actively growing shoots of conventionally raised plants were cultured on nutrient medium with the mineral salts and vitamins of Murashige and Skoog medium at 1/4 strength, supplemented with various concentrations of sucrose and 6-benzyladenine (BAP). Shoot regeneration from explants of both young (2 years old) and adult (20 years old) mother plants were readily achieved in the medium supplemented with 0.04-0.09 M sucrose with or without BAP. As many as 60-65 of the nodal segments cultured formed shoots. Rooting of regenerated shoots was observed in 50 of the explants harvested from young plants, whereas 25 of the explants rooted when the nodal explants were harvested from adult plants. The best rooting induction was achieved on 1/4 strength MS medium with vermiculite as the substrate and supplemented with 1-1.5 IBA (indolebutyric acid) and 1-2 PPZ (3-methyl-1-phenyl-2 pyrazolin-5-one). Plantlets were successfully transferred to soil.(AU)

Acylglycerol synthesis in liver of type II diabetic rats fed a diet supplemented with either N-6 or N-3 fatty acids

Liver is one of the tissues most actively involved in triacylglycerol synthesis and secretion. Hypertriglyceridemia is commonly associated with the diabetic state which has been detected in very young rats after the induction of experimental diabetes. In the present work, acylglycerol synthesis in liver of streprozotocintreated rats, fed a diet supplemented with n-3 and n-6 fatty acids, was studied. At the onset of the experiment, plasma triacylglycerol levels increased significantly in diabetic animals when compared to controls. Two weeks after the dietary treatment, the aforementoined parameter decreased in diabetic animals consuming either n-6 or n-3 fatty acids. In control rats, n-3 fatty acids depressed triacyglycerol synthesis in liver microsomes. In the diabetic group both diets increased diacylglycerol and triacylglycerol synthesis. The addition on liver cytosolic fraction from control rats to the incubation medium, stimulated the triacylglycerol synthesis in all the groups. Nevertheless, the radioactivity recovered in the neutral lipid fractions was lower in the samples from rats fed n-3 fatty acids compared to n-6. We conclude that dietary n-3 fatty acids decreased significantly triacylglycerol plasma levels in diabetic rats probably through the inhibiton of liver triacylglycerol secretion. In addition, there probably is an n-3 fatty sensitive factor in the liver cytosolic fraction able to depress triglyceride synthesis. (AU)

Efecto de la adición de lectina específica sobre la simbiosis Rhizobium leguminosarum--Phaseolus vulgaris / Effect of the addition of specific lectin on the symbiosis of Rhizobium leguminosarum and Phaseolus vulgaris

The effect of specific lectin addition on Rhizobium leguminosarum-Phaseolus vulgaris symbiosis characteristics was studied. Two alternatives were selected for comparison: a) P. vulgaris seedling roots treated with lectin were inoculated with R. leguminosarum and b) P. vulgaris seedling roots were inoculated with R. leguminosarum incubated with lectin for 3 h and 72 h. The following parameters were evaluated: number and dry weight of nodules and dry matter and nitrogen content of shoot. In lectin treatments, the weight of nodules (from 13 to 35), dry matter (from 6 to 18) and nitrogen content (from 5 to 28) increased significantly. The results suggest a stimulation in the formation of functional-nodules, specially in root (a) and Rhizobium (b) 72 h treatments. That is consistent with a model in which the lectin functions as an extracellular matrix component of the root interacting with roots and Rhizobium s.p. receptors.(AU)

Efecto de los acidos grasos poliinsaturados n-3 PUFA sobre el timo de ratas con deplecion proteica severa / Effect of n-3 PUFA plyunsatured fatty acids on the thymus of rats submetted to severe protein depletion

Se estudia el efecto de la malnutrición proteica severa y la posterior recuperación nutricional sin y con el agregado de (n-3) PUFA, sobre el timo de ratas en período de crecimiento activo. Se determina recuento celular, población T total y actividad de adenosina deaminasa (ADA) y purina nucleósido fosforilasa (PNP). La deprivación proteica several al destete provova un frenado en la proliferación y maduración celular del timo junto con el aumento de la actividad de las enzimas ADA y PNP. La administración de la dieta de caseína al 20 por ciento durante 9 días, sólo fue suficiente para revertir el efecto observado sobre la actividad de las enzimas estudiadas. La suplementación con 24mg/d de n-3 PUFA permite restablecer la proliferación y la maduración celular tímica. (AU)

Norepinephrine: stimulated activity of hypothalamic adenylate cyclase varies throughout the estrous cycle of the female rat

The activity of hypothalamic adenylate cyclase was studied throughout the estrous cycle of the female rat. The activity of the enzyme was determined in particulate fractions obtained from hypothalami of rats killed at 10.00 h and 16.00 h of the 4-day estrous cycle. The activity was assayed in the presence of norepinephrine (10(-8) to 10(-3) M) by the capacity to produce adenosine 3,5 cyclic monophosphate. The basal activity of adenylate cyclase was higher in the morning of estrus than at any other time during the cycle. Norepinephrine-stimulated adenylate cyclase activity, as assessed by the apparent affinity (Kd) and apparent maximum effect, varied during the cycle, showing highest affinity, lowest Kd, in the afternoon of proestrus. The highest level of apparent maximum effect was also found in the afternoon of proestrus declining on diestrous day 2, diestrous day 1 and estrus. The norepinephrine stimulated activity was significantly inhibited by phenoxybenzamine, an alpha-blocker, in the morning of diestrus day 1, whereas on the day of diestrus day 2 and proestrus it was inhibited by the beta-adrenoblocker, propranolol. A similar degree of inhibition by alpha- and beta-blockers was observed in the morning of estrus. These results indicate that the hypothalamic adenylate cyclase coupled to adrenergic receptors shows dynamic changes throughout the estrous cycle (Au)