Regeneration of the Corneal Epithelium after Debridement of its Central Region: An Autoradiographic Study on Rabbits

Purpose: To investigate the proliferative behavior of the corneal and limbal epithelia after debridement on the central region of the rabbit cornea. Methods: After scraping a circular epithelial area, 5 mm in diameter, in the center of the cornea, (3)H-thymidine ((3)H-TdR) was injected intravitreally, and the rabbits killed from 1 to 49 days afterward. The cornea, together with the adjacent conjunctiva, was processed for autoradiography. Results: The regenerating epithelium at the center of the cornea exhibited high frequencies of labeled nuclei when compared to controls. The mitotic indexes for the limbus were comparable in experimental and control eyes. The unique basal stratum of the limbal epithelium exhibited quick proliferation and vertical migration in all eyes. Cells that remained labeled for four weeks or more were observed throughout the corneal epithelium, including its basal stratum, and this did not depend on epithelial damage. Conclusion: Corneal epithelium wounds are healed by sliding and proliferation of cells surrounding the epithelial gap without any evidence for the participation of the limbal epithelium. Daughter cells labeled with (3)H-TdR were visualized in all layers of the corneal epithelium up to 7 weeks after the DNA precursor injection. However, at this long interval, the only labeled cells in the limbus were in the suprabasal layers.

End-systolic pressure-diameter relation of the left ventricle during transient and sustained elevations of blood pressure

OBJECTIVE: To assess the effect of transient and sustained variations in cardiac load on the values of the end-systolic pressure-diameter relation (ESPDR) of the left ventricle. METHODS: We studied 13 dogs under general anesthesia and autonomic blockade. Variations of cardiac loads were done by elevation of blood pressure by mechanical constriction of the aorta. Two protocols were used in each animal: gradual peaking and decreasing pressure variation, the transient arterial hypertension protocol (TAH), and a quick and 10 min sustained elevation, the sustained arterial hypertension protocol(SAH). Then, we compared the ESDR in these two situations. RESULTS: Acute elevation of arterial pressure, being it transitory or sustained, did not alter the heart frequency and increased similarly the preload and after load. However, they acted differently in end systolic pressure-diameter relation. It was greater in the SAH than TAH protocol, 21.0±7.3mmHg/mm vs. 9.2±1.2mmHg/mm (p<0.05). CONCLUSION: The left ventricular ESPDR values determined during sustained pressure elevations were higher than those found during transient pressure elevations. The time-dependent activation of myocardial contractility associated with the Frank-Starling mechanism is the major factor in inotropic stimulation during sustained elevations of blood pressure, determining an increase in the ESPDR values.

Community structure and comparative analysis of the woody component of a cerrado remnant in Southeastern Brazil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Anti-inflammatory and analgesic activities of the crude extracts from stem bark of Bauhinia guianensis

Methylene chloride, ethyl acetate and methanolic extracts front the stem bark of Bauhinia guianensis (Leguminosae, Caesalpinoideae) were obtained. These extracts were evaluated for antiinflammatory activity which was conducted using carrageenin, dextran and histamine-induced paw edema in rats. The extracts of B. guianensis were also assessed for analgesic activity which was conducted using the writhing test in mouse. The different animal groups were treated with these extracts (100 mg/kg i.p. and p.o, IC50) 30 min prior to the application of stimuli. The methanolic extract demonstrated significant inhibition in the carrageenin-induced edema model. In the dextran-induced edema model, all three extracts inhibited the inflammatory process significantly with the methanolic extract being the most active. The ethyl acetate extract was the only one shown to be effective in the histamine-induced edema model. Finally all extracts inhibited effectively the algogenic process in the writhing test induced by acetic acid.

Characterization of Neural Stem/Progenitor Cells Expressing VEGF and its Receptors in the Subventricular Zone of Newborn Piglet Brain

Neural stem/progenitor cell (NSP) biology and neurogenesis in adult central nervous system (CNS) are important both towards potential future therapeutic applications for CNS repair, and for the fundamental function of the CNS. In the present study, we report the characterization of NSP population from subventricular zone (SVZ) of neonatal piglet brain using in vivo and in vitro systems. We show that the nestin and vimentin-positive neural progenitor cells are present in the SVZ of the lateral ventricles of neonatal piglet brain. In vitro, piglet NSPs proliferated as neurospheres, expressed the typical protein of neural progenitors, nestin and a range of well-established neurodevelopmental markers. Upon dissociation and subculture, piglet NSPs differentiated into neurons and glial cells. Clonal analysis demonstrates that piglet NSPs are multi-potent and retain the capacity to generate both glia and neurons. These cells expressed VEGF, VEGFR1, VEGFR2 and Neuropilin-1 and -2 mRNAs. Real time PCR revealed that SVZ NSPs from newborn piglet expressed total VEGF and all VEGF splice variants. These findings show that piglet NSPs may be helpful to more effectively design growth factor based strategies to enhance endogenous precursor cells for cell transplantation studies potentially leading to the application of this strategy in the nervous system disease and injury.

Effect of calcium and magnesium silicate on the growth of the castor oil plant subjected to salinity levels

Salt stress decreases the osmotic potential of soil solution causing water stress, causing toxic effects in the plants resulting in injuries on the metabolism and nutritional disorders, thus compromising the plant growth, resulting in lower production. The calcium silicate and magnesium can perform the same function as limestone, besides providing silicon to plants, may also contribute to the resistance of plants to salt stress. Thus, the objective of this study was to evaluate the effect of calcium and magnesium silicate on the growth of the castor oil plant BRS Energia cultivated under saline conditions. This study evaluated plant height, stem diameter, number of leaves, leaf area, dry weight of shoot and root, and soil chemical characteristics. There was no interaction between factors of salinity level and of silicate level regarding the evaluated variables. There was a direct relationship between salinity levels and plant growth in height and stem diameter. The K concentration in soil were affected by salinity levels.

Clinical outcome in chronic myeloid leukemia after allogeneic hematopoietic stem cell transplantation: The experience of the Bone Marrow Transplantation Unit of FUNFARME/BRAZIL using FISH

Investigation of the efficacy of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in chronic myeloid leukemia patients is essential to predict prognosis and survival. In 20 patients treated at the Bone Marrow Transplantation Unit of São José do Rio Preto (São Paulo, Brazil), we used fluorescence in situ hybridization (FISH) to investigate the frequency of cells with BCR/ABL rearrangement at diagnosis and at distinct intervals after allo-HSCT until complete cytogenetic remission (CCR). We investigated the disease-free survival, overall survival in 3 years and transplant-related mortality rates, too. Bone marrow samples were collected at 1, 2, 3, 4, 6, 12, and 24 months after transplantation and additional intervals as necessary. Success rate of the FISH analyses was 100%. CCR was achieved in 75% of the patients, within on average of 3.9 months; 45% patients showed CCR within 60 days after HSCT. After 3 years of the allo-HSCT, overall survival rate was 60%, disease-free survival was 50% and the transplant-related mortality rate was 40%. The study demonstrated that the BCR-ABL FISH assay is useful for follow-up of chronic myeloid leukemia patients after HSCT and that the clinical outcome parameters in our patient cohort were similar to those described for other bone marrow transplantation units. ©FUNPEC-RP.

Compatibility of grafting in the melon cv. bônus No. 2, and effects on production

Twelve rootstocks were evaluated: 1) pumpkin 'Big Power' (Cucurbita ntoschata); 2) pumpkin 'Seca' (Cucurbita moschata); 3) bottle gourd 'Longa' (Lagenaria siceraria); 4) 'Cachi' (Lagenaria siceraria); 5) bottle gourd 'Marimba' (Lagenaria siceraria); 6) 'Mogango' (Cucurbita maxima); 7) pumpkin 'Kirameki' (Cucurbita moschata); 8) pumpkin 'Caravela' (Cucurbita moschata); 9) pumpkin 'Shelper' (Cucurbita moschata); 10) 'Gherkin' (Cucumis anguria); 11) 'Loofah' (Luffa cylindrica); and 12) pumpkin 'Goianinha' (Cucurbita moschata) with respect to compatibility with melon yield, and the production and quality of fruits from cv. Bônus No. 2, non-grafted and grafted with 9 of these rootstocks (1, 2, 4, 5, 6, 7, 9, 10 and 11). The bottle gourd 'Marimba' provided the highest percentage of grafting success between the vine and rootstock. The rootstocks 1, 2, 3, 4 and 7 did not differ from bottle gourd 'Marimba', therefore also indicating good compatibility with the melon cv. Bônus No. 2. For height of the plants, it was shown in the first evaluation that rootstock 5 produced a greater height of the plant, differing only from rootstocks 7, 8, 9, 10 and 11 and non-grafted cv. Bônus No. 2. In the second evaluation, 'Big Power' showed the greatest value for height of the plant, differing only from combinations with 'Cachi', 'Mogango', 'Shelper' and 'Loofah'. With regard to number of leaves, in the first evaluation rootstocks 3, 4 and 6 had the greatest number of leaves, but in the second 4, 9 and 11 had the greatest. For dry weight of the stem, the greatest value was obtained with 'Mogango' in the first evaluation and with 'Big Power' in the second. For leaf area of the plants, a difference was found among the treatments only in the first evaluation, where the combination with bottle gourd 'Longa' showed a greater leaf area, but did not differ from the combinations with rootstocks 1, 4, 5 and 6. Differences were demonstrated among the treatments only for the transverse diameter of the fruit, where the combination with rootstock bottle gourd 'Marimba' showed the greatest value, differing only from the combination with 'Gherkin'. There were no significant differences for the mean longitudinal diameter, pulp thickness and total soluble solids among the treatments studied.

Influence of the implant diameter with different sizes of hexagon: Analysis by 3-dimensional finite element method

The aim of this study was to evaluate the stress distribution in implants of regular platforms and of wide diameter with different sizes of hexagon by the 3-dimensional finite element method. We used simulated 3-dimensional models with the aid of Solidworks 2006 and Rhinoceros 4.0 software for the design of the implant and abutment and the InVesalius software for the design of the bone. Each model represented a block of bone from the mandibular molar region with an implant 10 mm in length and different diameters. Model A was an implant 3.75 mm/regular hexagon, model B was an implant 5.00 mm/regular hexagon, and model C was an implant 5.00 mm/ expanded hexagon. A load of 200 N was applied in the axial, lateral, and oblique directions. At implant, applying the load (axial, lateral, and oblique), the 3 models presented stress concentration at the threads in the cervical and middle regions, and the stress was higher for model A. At the abutment, models A and B showed a similar stress distribution, concentrated at the cervical and middle third; model C showed the highest stresses. On the cortical bone, the stress was concentrated at the cervical region for the 3 models and was higher for model A. In the trabecular bone, the stresses were less intense and concentrated around the implant body, and were more intense for model A. Among the models of wide diameter (models B and C), model B (implant 5.00 mm/regular hexagon) was more favorable with regard to distribution of stresses. Model A (implant 3.75 mm/regular hexagon) showed the largest areas and the most intense stress, and model B (implant 5.00 mm/regular hexagon) showed a more favorable stress distribution. The highest stresses were observed in the application of lateral load.

Alkaloids and volatile constituents from the stem of Fusaea longifolia (Aubl.) Saff. (Annonaceae)

ABSTRACT: A phytochemical study of the ethanol extract and an extraction of the volatile compounds, performed by means of Clevenger apparatus were carried out with the stem of Fusaea longifolia (Aubl.) Saff. (Annonaceae). The ethanol extract yielded O-methylmoschatoline, isolated for the first time in this species, and stepholidine, reported for the first time in genus Fusaea. The structural identification of the alkaloids was made based on the analysis of their NMR spectra. Through the use of GC and GC-MS, two sesquiterpenoids, a-cadinol (12.5%) and spatulenol (12.0%) were identified as the major constituents of the essential oil.

Bone Marrow'S Harvest in The Coxal Tuberosity for Isolation and Culture of Mesenchymal Stem Cells of Buffaloes (Bubalus Bubalis)

Several studies with mesenchymal stem cells (MSCs) have been developed in many species because of its ability to differentiate into other mesoderm lineages, capacity of self-regeneration, low immunogenicity, paracrine, anti-inflamatory, immunomodulatory and antiapoptotic effects which make then a promissory source to be used in therapeutic strategies. The aim of this study is to report the technique of harvest of bone marrow (BM) in the coxal tuberosity (CT) of buffaloes. For this, the animals were selected, identified and contained in a stock. Then trichotomy was performed in the region corresponding to the CT. After identifying the anatomic site it was performed antisepsis, local anesthetic block and introduction of a myelogram's needle (Lang(R)) for BM aspiration. Once the needle was firmly fixed in the CT, the mandril was removed and proceeded to BM aspiration with a syringe (20 mL) containing 1 ml of heparin at 1000 IU / mL and 1 mL of PBS. After the collection, each sample collected was manually homogenized, identified and referred to the LRACT - FMVZ / UNESP-BRAZIL for the correct processing. The anatomical site tested showed to be an alternative site of harvest of BM once provided the appropriate isolation and culture of the mononuclear fraction. Moreover, the procedure was performed without difficulty and with great security. Based on this, it can be conclude that CT is an excellent anatomical site for isolation and culture of MSCs and the proposed technique is viable and feasible to be held in buffaloes.

Comparative wood anatomy of root and stem of Citharexylum myrianthum (Verbenaceae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)