Plasticity of protein expression during culture of fetal skin cells.

In order to gain insight into the biology of fetal skin during culture, cellular proteins were studied during four culture passages (P00, P01, P04 as well as P10) using high-resolution two-dimensional (2-D) gel electrophoresis and mass spectrometry (MS). Bioinformatic analyses were focused on a region of each gel corresponding to pI between 4 and 8 and M(r) from 8000 to 35 000. In this area, 373 +/- 42 spots were detected (N = 18). Twenty-six spots presented an integrated intensity that increased in the higher passages, whereas five spots showed a progressively lower intensity in subsequent passaging. MS analysis was performed on spots that were unambiguously identified on preparative 2-D gels. Among the 26 spots showing an increased size between P00 and P10, 9 were identified, and corresponded to 3 proteins: (i) peptidyl-prolyl cis-trans isomerase A (P05092; cyclophilin A or cyclosporin A-binding protein), (ii) triosephosphate isomerase (P00938), and (iii) enoyl-CoA hydratase (P30084). Among these nine identified spots, three were absent at P00, but were present at P10. They corresponded to isoforms of peptidyl-prolyl cis-trans isomerase and triosephosphate isomerase, respectively. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyses of the acidic isoforms of triosephosphate isomerase showed modifications of cysteine residues to cysteic acid. All these isoforms were clearly present in the skin cells of a 4-year-old child, as well as in skin cells from a 80-year-old man, at P00. These observations probably reflect either an oxidative stress related to cell culture, or, alternatively, maturation, differentiation and the aging of the cells.

Macronutrient uptake, accumulation and export by the irrigated 'vitória' pineapple plant

The nutritional state of the pineapple plant has a large effect on plant growth, on fruit production, and fruit quality. The aim of this study was to assess the uptake, accumulation, and export of nutrients by the irrigated 'Vitória' pineapple plant during and at the end of its development. A randomized block statistical design with four replications was used. The treatments were defined by different times of plant collection: at 270, 330, 390, 450, 510, 570, 690, 750, and 810 days after planting (DAP). The collected plants were separated into the following components: leaves, stem, roots, fruit, and slips for determination of fresh and dry matter weight at 65 ºC. After drying, the plant components were ground for characterization of the composition and content of nutrients taken up and exported by the pineapple plant. The results were subjected to analysis of variance, and non-linear regression models were fitted for the significant differences identified by the F test (p<0.01). The leaves and the stem were the plant components that showed the greatest accumulation of nutrients. For production of 72 t ha-1 of fruit, the macronutrient accumulation in the 'Vitória' pineapple exhibited the following decreasing order: K > N > S > Ca > Mg > P, which corresponded to 898, 452, 134, 129, 126, and 107 kg ha-1, respectively, of total accumulation. The export of macronutrients by the pineapple fruit was in the following decreasing order: K > N > S > Ca > P > Mg, which was equivalent to 18, 17, 11, 8, 8, and 5 %, respectively, of the total accumulated by the pineapple. The 'Vitória' pineapple plant exported 78 kg ha-1 of N, 8 kg ha-1 of P, 164 kg ha-1 of K, 14 kg ha-1 of S, 10 kg ha-1 of Ca, and 6 kg ha-1 of Mg by the fruit. The nutrient content exported by the fruits represent important components of nutrient extraction from the soil, which need to be restored, while the nutrients contained in the leaves, stems and roots can be incorporated in the soil within a program of recycling of crop residues.

Iowa History and Culture : A Bibliography of Materials Published Between 1952 and 1986, 1989

This bibliography was compiled by two reference librarians, Patricia Dawson and David Hudson with the goal of making it easier of tracking down material on Iowa history and culture. This supplements the Iowa History Reference Guide published in 1952 by William Petersen.

Peroxynitrite cytotoxicity on bovine retinal pigmented epithelial cells in culture.

Peroxynitrite induced in vitro a dose dependent toxicity on retinal pigmented epithelial (RPE) cells. Cell death was partially mediated by apoptosis as demonstrated by nuclear fragmentation and TdT-mediated dUTP nick-end labeling assay. Peroxynitrite-induced tyrosine nitration was revealed by immunocytochemistry, both in the cytoplasm and in the nucleus of the cells. Nitration was not observed in RPE cells, producing nitric oxide (NO) after stimulation by lipopolysacharide and interferon-g (IFN-gamma), suggesting that peroxynitrite was not formed in vitro in such conditions. Peroxynitrite could be responsible for the retinal damages observed in pathological conditions in which NO has been demonstrated to be involved. In this context, EGb761, identified as a free radical scavenger, was showed herein to protect RPE cells against peroxynitrite injury.

ORGANIC MATTER FRACTIONS IN A QUARTZIPSAMMENT UNDER CULTIVATION OF IRRIGATED MANGO IN THE LOWER SÃO FRANCISCO VALLEY REGION, BRAZIL

Improper land use has lead to deterioration and depletion of natural resources, as well as a significant decline in agricultural production, due to decreased soil quality. Removal of native vegetation to make way for agricultural crops, often managed inadequately, results in soil disruption, decreased nutrient availability, and decomposition of soil organic matter, making sustainable agricultural production unviable. Thus, the aim of the present study was to evaluate the impact of growing irrigated mango (over a 20 year period) on the organic carbon (OC) stocks and on the fractions of soil organic matter (SOM) in relation to the native caatinga (xeric shrubland) vegetation in the Lower São Francisco Valley region, Brazil. The study was carried out on the Boa Esperança Farm located in Petrolina, Pernambuco, Brazil. In areas under irrigated mango and native caatinga, soil samples were collected at the 0-10 and 10-20 cm depths. After preparing the soil samples, we determined the OC stocks, carbon of humic substances (fulvic acid fractions, humic acid fractions, and humin fractions), and the light and heavy SOM fractions. Growing irrigated mango resulted in higher OC stocks; higher C stocks in the fulvic acid, humic acid, and humin fractions; and higher C stocks in the heavy and light SOM fraction in comparison to nativecaatinga, especially in the uppermost soil layer.

The long-term culture of porcine urothelial cells and induction of urothelial stratification.

OBJECTIVE: To assess porcine urothelial cell cultures and the in vitro induction of urothelial stratification in long-term cultures, to study their morphological, functional and genetic behaviour, and thus provide potential autologous urothelium for tissue-engineered substitutes for demucosalized gastric or colonic tissue. MATERIALS AND METHODS: Primary cultures of porcine urothelium were established and the cells passaged thereafter. Cell specificity was confirmed by cytokeratin analysis, cell membrane stability assessed using lactate dehydrogenase leakage, cell de-differentiation by gamma-glutamyl transferase activity and genomic stability by karyotype investigations. Histology and scanning electron microscopy were performed to study the cultured cells and the stratified constructs. Furthermore, collagen matrices were tested as cell scaffolds. RESULTS: The cells were cultured for 180 days; 10 subcultures were established during this period. Stratification was induced in a culture flask and on a collagen matrix. Cytokeratins 7, 8, 17 and 18 were expressed in all cultures, and cell membranes were stable, with no evident de-differentiation. The cultures were stable in their genotype and no chromosomal aberrations were found. The histology and immunohistochemistry of the stratified porcine constructs, and cell membrane stability and cell de-differentiation, were compared with those in the human system. CONCLUSION: Pig and human urothelial cells can be cultured over a long period with no signs of senescence. Urothelial stratification can be induced in vitro. The collagen matrix seems to be an excellent scaffold, allowing cell adherence and growth.

Water Balance Components in Covered and Uncovered Soil Growing Irrigated Muskmelon

ABSTRACT Knowledge of the terms (or processes) of the soil water balance equation or simply the components of the soil water balance over the cycle of an agricultural crop is essential for soil and water management. Thus, the aim of this study was to analyze these components in a Cambissolo Háplico (Haplocambids) growing muskmelon (Cucumis melo L.) under drip irrigation, with covered and uncovered soil, in the municipality of Baraúna, State of Rio Grande do Norte, Brazil (05º 04’ 48” S, 37º 37’ 00” W). Muskmelon, variety AF-646, was cultivated in a flat experimental area (20 × 50 m). The crop was spaced at 2.00 m between rows and 0.35 m between plants, in a total of ten 50-m-long plant rows. At points corresponding to ⅓ and ⅔ of each plant row, four tensiometers (at a distance of 0.1 m from each other) were set up at the depths of 0.1, 0.2, 0.3, and 0.4 m, adjacent to the irrigation line (0.1 m from the plant row), between two selected plants. Five random plant rows were mulched using dry leaves of banana (Musa sp.) along the drip line, forming a 0.5-m-wide strip, which covered an area of 25 m2 per of plant row with covered soil. In the other five rows, there was no covering. Thus, the experiment consisted of two treatments, with 10 replicates, in four phenological stages: initial (7-22 DAS - days after sowing), growing (22-40 DAS), fruiting (40-58 DAS) and maturation (58-70 DAS). Rainfall was measured with a rain gauge and water storage was estimated by the trapezoidal method, based on tensiometer readings and soil water retention curves. For soil water flux densities at 0.3 m, the tensiometers at the depths of 0.2, 0.3, and 0.4 m were considered; the tensiometer at 0.3 m was used to estimate soil water content from the soil water retention curve at this depth, and the other two to calculate the total potential gradient. Flux densities were calculated through use of the Darcy-Buckingham equation, with hydraulic conductivity determined by the instantaneous profile method. Crop actual evapotranspiration was calculated as the unknown of the soil water balance equation. The soil water balance method is effective in estimating the actual evapotranspiration of irrigated muskmelon; there was no significant effect of soil coverage on capillary rise, internal drainage, crop actual evapotranspiration, and muskmelon yield compared with the uncovered soil; the transport of water caused by evaporation in the uncovered soil was controlled by the break in capillarity at the soil-atmosphere interface, which caused similar water dynamics for both management practices applied.

A simple blood-culture bacterial pellet preparation for faster accurate direct bacterial identification and antibiotic susceptibility testing with the VITEK 2 system.

An ammonium chloride procedure was used to prepare a bacterial pellet from positive blood cultures, which was used for direct inoculation of VITEK 2 cards. Correct identification reached 99% for Enterobacteriaceae and 74% for staphylococci. For antibiotic susceptibility testing, very major and major errors were 0.1 and 0.3% for Enterobacteriaceae, and 0.7 and 0.1% for staphylococci, respectively. Thus, bacterial pellets prepared with ammonium chloride allow direct inoculation of VITEK cards with excellent accuracy for Enterobacteriaceae and a lower accuracy for staphylococci.