A simple procedure to obtain one-micrometer sections of routinely embedded paraffin material

By freezing blocks of paraffin-embedded tissues to a convenient temperature it is possible to obtain routinely 1 micron sections that can be further processed as normal thicker sections. Normal and disposable steel knives can be used and the staining time should be increased in most procedures. Gradual freezing of blocks to the temperature of dry ice is the simplest and safest way to obtain an adequate temperature. The best results were obtained using as fixative 4% paraformaldehyde in phosphate buffered saline solution.     

Effects of 6-hydroxydopamine and 5-hydroxydopamine onlthe teleost melanophores innervation

Intraperitoneal injections of 6-OH-dopamine (80 mg kg^−l) promote, in toad fish, killifish, lsummer and winter flounders, a darkening of their colour and loss of capacity to adapt tolthe colour of the background. This condition persisted for three weeks after which thelanimals gradually returned to normal. The study of the skin of 6-hydroxydopamineltreated killifish showed degenerative lesions in the fine nerves and synapses of its melanophores, 124 h after the administration of the drug. These lesions progressed and 4 days later, lno synaptic structures could be detected in these cells. This condition persisted up to thel20th day. These results suggest that melanophores have a single monoaminergic innervaion.     

Structure of the tail fin in teleosts

The influence of adrenalectomy and administration of hypertonic saline on the amount of vasopressin, oxytocin, and neurophysin contained in the median eminence and the neural lobe of rats was studied by means of the following methods: (i) morphometric and microphotometric analyses of aldehyde fuchsin-stained histological sections of the neurohypophysis; (ii) immunohistochemical demonstration of vasopressin, oxytocin, and neurophysin in the neurohypophysis, and (iii) radioimmunological measurement of vasopressin and oxytocin in extracts of the median eminence and the neural lobe. Adrenalectomy increases the amount of vasopressin and neurophysin in the external layer of the median eminence but does not change the content of oxytocin. It has no influence on the amount of vasopressin, oxytocin, and neurophysin demonstrable in the inner layer of the median eminence and in the neural lobe two weeks after the operation. Hypertonic saline markedly diminishes the vasopressin, oxytocin, and neurophysin content of the inner layer of the median eminence and the neural lobe but reduces only slightly, if at all, the amount of vasopressin and neurophysin in the outer layer of the median eminence. The findings support the concept that osmotic stress reduces only the vasopressin and oxytocin content of the hypothalamus-neural lobe system and has no or only little influence on the vasopressin content of the outer layer of the median eminence.     

Functional Activity of Blood Polymorphonuclear Leukocytes as an Oxidative Stress Biomarker in Human Subjects

In the present work, we studied the role of polymorphonuclear leukocytes (PMN) in aged individuals and coronary heart disease (CHD)-bearing patients, two physiopathological processes associated with overproduction of reactive oxygen species (ROS). The effects of antioxidant supplementation on the functional activity of PMN from CHD patients were also determined. The function of PMNs was evaluated by measuring of phagocytosis, killing activity, and ROS production. Luminol amplified chemiluminescence (CL) was used to estimate ROS production by stimulated PMNs. Total cholesterol and the LDL-cholesterol fraction from CHD patients were found to be higher than those recommended, returning to normal levels after antioxidant therapy. PMN CL of CHD patients was found to be higher than the associated control groups. Antioxidant therapy administrated to CHD patients lead to an increase in the killing activity accompanied by a decrease in PMN CL of these subjects. The study also showed that killing activity of PMN from human subjects over 60 years was significantly lower than the activity measured in younger subjects. PMN CL produced after stimulation was found to be positively correlated with the increasing age of human subjects (r = .946, p < .01).     

Chronic fenfluramine treatment of rats with different ages: Effects on brain oxidative stress-related parameters

Fenfluramine is an anorectic drug widely used for the regulation of food intake that presents some adverse effects at the central and peripheral levels. d-Fenfluramine, an isomer of dl-fenfluramine, is postulated to be more effective and to induce less side effects than the racemic compound. These drugs act preferentially on the serotonergic system. Some authors have suggested that fenfluramine causes a degeneration of serotonergic neurons. Alterations of the serotonergic system are also observed during the aging process, and in this case, a relationship with reactive oxygen species has been already established. In view of these data, the present study was conducted to investigate the relationship between fenfluramine and brain antioxidant defense system in mature and aged animals. Rats aged 4 and 17 months were chronically treated with dl-fenfluramine, d-fenfluramine, or saline. Brain activity of superoxide dismutase and glutathione peroxidase was significantly affected by aging. Catalase activity was altered by the treatment. Total glutathione content and chemiluminescence in the brains were also altered by aging. Glutathione levels were altered as a function of the interaction between age and treatment. These findings suggest that treatment with d- or dl-fenfluramine results in alteration of the antioxidant system that could be exacerbated when associated with the aging process. © 1997 John Wiley & Sons, Inc.     

Effect of DHEA Glutamate Release from Synaptosomes of Rats at Different Ages

Dehydroepiandrosterone (DHEA) exerts multiple effects in the central nervous system. Most of them seem to be mediated through their nongenomic actions on neurotransmitter receptors, and these actions occur within seconds or milliseconds. DHEA increases neuronal excitability, enhances neuronal plasticity, and has neuroprotective properties. By investigating glutamate release from synaptosomes of rats at different ages (from 17 days to 12 months), we observed that (i) there is an increase in basal and K(+)-stimulated L-[3H] glutamate release in rats at 12 months old, when compared to other ages; and (ii) there is an inhibitory effect of DHEA on basal L-[3H] glutamate release in 12 months old. This inhibitory effect of DHEA could be related to its reported protective role against excitotoxicity caused by overstimulation of the glutamatergic system and ageing.     

Fungal flora of the digestive tract of 5 species of triatomines vectors of Trypanosoma cruzi,Chagas 1909

A study of the mycobiota in the digestive tract of 5 important species of triatomines, Triatoma brasiliensis, T. infestans, T. sordida, T. pseudomaculata and T. vitticeps, was made. The digestive tracts of 164 adults and 535 nymphs of those triatomines were studied and 393 fungal strains were isolated.The genera with the greatest number of species were Penicillium (19 species), Aspergillus (17 species) and Acremonium (5 species) and the most frequent species, in decreasing order, were Penicillium corylophilum, Aspergillus niger, Penicillium fellutanum, Cladosporium herbarum, Penicillium waksmanii, Aspergillus awamori and Paecilomyces variotii. Among the isolated fungi, we found species that are recognized as entomopathogenic and pathogenic for humans and animals.This revised version was published online in October 2005 with corrections to the Cover Date.     

A systematic RNAi synthetic interaction screen reveals a link between p53 and snoRNP assembly

TP53 (tumour protein 53) is one of the most frequently mutated genes in human cancer and its role during cellular transformation has been studied extensively. However, the homeostatic functions of p53 are less well understood. Here, we explore the molecular dependency network of TP53 through an RNAi-mediated synthetic interaction screen employing two HCT116 isogenic cell lines and a genome-scale endoribonuclease-prepared short interfering RNA library. We identify a variety of TP53 synthetic interactions unmasking the complex connections of p53 to cellular physiology and growth control. Molecular dissection of the TP53 synthetic interaction with UNRIP indicates an enhanced dependency of TP53-negative cells on small nucleolar ribonucleoprotein (snoRNP) assembly. This dependency is mediated by the snoRNP chaperone gene NOLC1 (also known as NOPP140), which we identify as a physiological p53 target gene. This unanticipated function of TP53 in snoRNP assembly highlights the potential of RNAi-mediated synthetic interaction screens to dissect molecular pathways of tumour suppressor genes.     

Secondary Forests on Anthropogenic Soils of the Middle Madeira River: Valuation,Local Knowledge,and Landscape Domestication in Brazilian Amazonia

Secondary Forests on Anthropogenic Soils of the Middle Madeira River: Valuation, Local Knowledge, and Landscape Domestication in Brazilian Amazonia. Anthropogenic forests and soils are widespread throughout Amazonia and are the product of the landscape domestication process carried out by Amazonian societies since pre-Colombian times. Areas of Terra Preta de índio (TPI, Amazonian Dark Earths) are recognized by local rural residents and associated with specific forms of use and management of these soils and associated secondary forests. We used a quantitative approach to investigate how secondary forests on TPI are recognized and used by local residents along the middle Madeira River, Central Amazonia. Sixty-two residents were interviewed in three riverside communities and listed the ethnospecies and their uses in secondary forests on TPI and on non-anthropogenic soils (NAS). Local residents mentioned more ethnospecies on TPI (mean ± standard deviation: 19.5 ± 8.9) than on NAS (17.4 ± 8.5), and the use value of the environment to the informants (UVia) was higher on TPI (19 ± 5.7) than on NAS (16.2 ± 6.0). Eleven ethnospecies were classified as anthropogenic soil indicators, among which three intensively used palms are widely recognized as indicators of anthropogenic areas and two are domesticated to some degree. The intimate and lasting interactions between humans and TPI have favored the maintenance of secondary forests in these domesticated landscapes with a diverse assemblage of useful and domesticated species. Rural residents in Amazonia recognize these forests as an important source of food and other resources. The use, management, and traditional knowledge related to these domesticated landscapes may provide useful information for the understanding of Amazonian historical ecology and for the design of more efficient biodiversity management and conservation plans.