Effect of desiccation on the ultrastructural appearances of Acinetobacter baumannii and Acinetobacter lwoffii.
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An Acinetobacter baumannii isolate survived desiccation beyond 30 days and an Acinetobacter lwoffii isolate up to 21 days. For both species, desiccation resulted in a significant increase in the proportion of round cells (A baumannii, 40% to 80%; A lwoffii, 51% to 63%) and a significant decrease in rod shaped cells (A baumannii, 58% to 13%; A lwoffii, 46% to 34%). Electronmicroscopic examination showed that there was also a corresponding significant increase in the cell wall thickness (A baumannii, up to 53%; A lwoffii, up to 26%). Desiccated A baumannii cells became more electron-dense and had significantly thicker cell walls (x1.3) than those of A lwoffii. Cell wall structures of A baumannii strains with different abilities to resist desiccation deserve further study. (+info)
Evaluation of dehydrated restaurant food waste products as feedstuffs for finishing pigs.
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Two dehydrated restaurant food waste (DFW) products were evaluated as potential feedstuffs for finishing pigs. For each product, fresh food wastes were obtained from food service operations at a resort complex in central Florida. The wastes were mostly leftover food and plate scrapings. The wastes were minced, blended with a feed stock (soy hulls and wheat flour [DFW1] or soy hulls and ground corn [DFW2]), pelleted, and dried. The dried product was then blended with additional minced fresh food wastes and dried; this process was then repeated. The final DFW products contained approximately 60% dried food wastes. The DFW1 and DFW2 products contained 11.4 and 8.4% moisture, 15.0 and 14.4% CP, 13.8 and 16.0% crude fat, 10.4 and 14.5% crude fiber, 5.8 and 4.7% ash, .63 and .64% lysine, .54 and .63% Ca, .34 and .38% P, .69 and .86% Cl, and .35 and .47% Na, respectively. Two feeding trials with 48 and 72 finishing pigs (56 to 112 kg), respectively, were conducted comparing diets without (control) or with the DFW product included at 40% of the diet (DFW1) for Trial 1 and 40 or 80% of the diet (DFW2) for Trial 2. Pigs fed the DFW diets in both trials had ADG that were similar (P > . 10) to and average gain:feed ratios that were superior (P = .06, Trial 1; P < .01; linear, Trial 2) to those for control pigs. Carcass lean content and lean quality scores were not reduced (P > . 10) by feeding pigs the DFW diets in either trial. Carcass fat became softer (P < .01; linear) with increasing amount of DFW2 in the diet in Trial 2. Thus, dehydrated restaurant food wastes have the potential to produce a nutritious feedstuff for pigs while offering a viable solid waste disposal option. (+info)
Dehydration and crystallization of trehalose and sucrose glasses containing carbonmonoxy-myoglobin.
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We report a study wherein we contemporarily measured 1) the dehydration process of trehalose or sucrose glasses embedding carbonmonoxy-myoglobin (MbCO) and 2) the evolution of the A substates in saccharide-coated MbCO. Our results indicate that microcrystallization processes, sizeably different in the two saccharides, take place during dehydration; moreover, the microcrystalline structure is maintained unless the dry samples are equilibrated with a humidity >/=75% (>/=60%) at 25 degrees C for the trehalose (sucrose) sample. The evolution of the parameters that characterize the A substates of MbCO indicates that 1) the effects of water withdrawal are analogous in samples dried in the presence or in the absence of sugars, although much larger effects are observed in the samples without sugar; 2) the distribution of A substates is determined by the overall matrix structure and not only by the sample water content; and 3) the population of A0 substate (i. e., the substate currently put in relation with MbCO molecules having the distal histidine out of the heme pocket) is largely enhanced during the dehydration process. However, after rehumidification its population is largely decreased with respect to the values obtained, at similar water content, during the first dehydration run. (+info)
Cell surface analysis techniques: What do cell preparation protocols do to cell surface properties?
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Cell surface analysis often requires manipulation of cells prior to examination. The most commonly employed procedures are centrifugation at different speeds, changes of media during washing or final resuspension, desiccation (either air drying for contact angle measurements or freeze-drying for sensitive spectroscopic analysis, such as X-ray photoelectron spectroscopy), and contact with hydrocarbon (hydrophobicity assays). The effects of these procedures on electrophoretic mobility, adhesion to solid substrata, affinity to a number of Sepharose columns, structural integrity, and cell viability were systematically investigated for a range of model organisms, including carbon- and nitrogen-limited Psychrobacter sp. strain SW8 (glycocalyx-bearing cells), Escherichia coli (gram-negative cells without a glycocalyx), and Staphylococcus epidermidis (gram-positive cells without a glycocalyx). All of the cell manipulation procedures severely modified the physicochemical properties of cells, but with each procedure some organisms were more susceptible than others. Considerable disruption of cell surfaces occurred when organisms were placed in contact with a hydrocarbon (hexadecane). The majority of cells became nonculturable after air drying and freeze-drying. Centrifugation at a high speed (15,000 x g) modified many cell surface parameters significantly, although cell viability was considerably affected only in E. coli. The type of washing or resuspension medium had a strong influence on the values of cell surface parameters, particularly when high-salt solutions were compared with low-salt buffers. The values for parameters obtained with different methods that allegedly measure similar cell surface properties did not correlate for most cells. These results demonstrate that the methods used to prepare cells for cell surface analysis need to be critically investigated for each microorganism so that the final results obtained reflect the nature of the in situ microbial cell surface as closely as possible. There is an urgent need for new, reliable, nondestructive, minimally manipulative cell surface analysis techniques that can be used in situ. (+info)
Regulation of the hepatocyte cell cycle by type I collagen matrix: role of cyclin D1.
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Rat hepatocytes adherent to a rigid film of type I collagen will spread and enter S phase, while those attached to collagen gel or a dried collagen substrate remain round and quiescent. The current studies were initiated to determine the mechanism by which these different substrates differentially influence cell cycle progression. Cyclin D1 mRNA and protein expression and associated kinase activity was low on dried collagen relative to collagen film. In contrast, cyclin E and cdk2 protein levels were similar on the two substrates. Although cyclin E and cdk2 were present, cells on dried collagen lacked cdk2 kinase activity. p27 protein levels did not differ between dried collagen and film, but more p27 was associated with cdk2 in cells on dried collagen than those on collagen film. Cyclin D1 expression on collagen film was inhibited by cytochalasin D and exoenzyme C3, suggesting a role for the GTP-binding protein, Rho, in regulating cyclin D1 expression. Cyclin D1 over-expression induced hepatocytes into S phase in the absence of cell shape change on dried collagen or collagen gel. These results demonstrate a novel, substrate-dependent mechanism for cyclin D1 expression in hepatocytes, and also demonstrate that cyclin D1 over-expression allows shape-independent S phase entry. (+info)
Isotope dilution spaces of mice injected simultaneously with deuterium, tritium and oxygen-18.
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The isotope dilution technique for measuring total body water (TBW), and the doubly labelled water (DLW) method for measuring energy expenditure, are both sensitive to small variations in the ratio of the hydrogen to oxygen-18 dilution space. Since the dilution space ratio varies between individuals, there has been much recent debate over what causes this variability (i.e. physiological differences between individuals or analytical error in the isotope determinations), and thus which values (individual or a population mean dilution space ratio) should be employed for TBW and DLW calculations. To distinguish between physiological and analytical variability, we injected 15 non-reproductive and 12 lactating mice (Mus musculus, outbred MF1) simultaneously with deuterium, tritium and oxygen-18. The two hydrogen labels were administered and analysed independently, therefore we expected a strong correlation between dilution space ratios based on deuterium and tritium if most of the variation in dilution spaces was physiological, but only a weak correlation if most of the variation was analytical. Dilution spaces were significantly influenced by reproductive status. Dilution spaces expressed as a percentage of body mass averaged 15.7 % greater in lactating mice than in non-reproductive mice. In addition, the hydrogen tracer employed had a significant effect (deuterium spaces were 2.0 % larger than tritium spaces). Deuterium and tritium dilution spaces, expressed as a percentage of body mass, were highly correlated. Dilution space ratios ranged from 0.952 to 1. 146 when using deuterium, and from 0.930 to 1.103 when using tritium. Dilution space ratios based on deuterium and tritium were also highly correlated. Comparison of standard deviations of the dilution space ratio based on deuterium in vivo and in vitro indicated that only 4.5 % of the variation in the dilution space ratios observed in the mice could be accounted for by analytical variation in the deuterium and oxygen-18 analyses. Although our results include data which were outside the limits previously regarded as biologically possible, the correlations that we detected strongly suggest that variation in the observed dilution space ratio was mostly physiological rather than analytical. (+info)
Active Fe-containing superoxide dismutase and abundant sodF mRNA in Nostoc commune (Cyanobacteria) after years of desiccation.
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Active Fe-superoxide dismutase (SodF) was the third most abundant soluble protein in cells of Nostoc commune CHEN/1986 after prolonged (13 years) storage in the desiccated state. Upon rehydration, Fe-containing superoxide disumutase (Fe-SOD) was released and the activity was distributed between rehydrating cells and the extracellular fluid. The 21-kDa Fe-SOD polypeptide was purified, the N terminus was sequenced, and the data were used to isolate sodF from the clonal isolate N. commune DRH1. sodF encodes an open reading frame of 200 codons and is expressed as a monocistronic transcript (of approximately 750 bases) from a region of the genome which includes genes involved in nucleic acid synthesis and repair, including dipyrimidine photolyase (phr) and cytidylate monophosphate kinase (panC). sodF mRNA was abundant and stable in cells after long-term desiccation. Upon rehydration of desiccated cells, there was a turnover of sodF mRNA within 15 min and then a rise in the mRNA pool to control levels (quantity of sodF mRNA in cells in late logarithmic phase of growth) over approximately 24 h. The extensive extracellular polysaccharide (glycan) of N. commune DRH1 generated superoxide radicals upon exposure to UV-A or -B irradiation, and these were scavenged by SOD. Despite demonstrated roles for the glycan in the desiccation tolerance of N. commune, it may in fact be a significant source of damaging free radicals in vivo. It is proposed that the high levels of SodF in N. commune, and release of the enzyme from dried cells upon rehydration, counter the effects of oxidative stress imposed by multiple cycles of desiccation and rehydration during UV-A or -B irradiation in situ. (+info)
Structural characterization of the released polysaccharide of desiccation-tolerant Nostoc commune DRH-1.
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The structure of the viscous extracellular polysaccharide (glycan) of desiccation-tolerant Nostoc commune DRH-1 was determined through chromatographic and spectroscopic methods. The polysaccharide is novel in that it possesses a 1-4-linked xylogalactoglucan backbone with D-ribofuranose and 3-O-[(R)-1-carboxyethyl]-D-glucuronic acid (nosturonic acid) pendant groups. The presence of D-ribose and nosturonic acid as peripheral groups is unusual, and their potential roles in modulating the rheological properties of the glycan are discussed. Nosturonic acid was present in the glycans of N. commune from diverse geographic locations, suggesting that this uronic acid is an integral component of this cosmopolitan anhydrophile. (+info)