Registration of confocal scanning laser microscopy and quantitative backscattered electron images for the temporospatial quantification of mineralization density in 18-month old thoroughbred racehorse articular calcified cartilage

Combined backscattered electron scanning electron microscopy (BSE SEM) and confocal scanning laser microscopy (CSLM) have been used to put tissue mineralization data into the context of soft tissue histology and fluorescent label information. Mineralization density (Dm) and linear accretion rate (LAR) are quantifiable parameters associated with mineralizing fronts within calcified tissues. Quantitative BSE (qBSE) may be used to determine Dm, while CSLM may be used to detect label fluorescence from which LAR is calculated. Eighteen-month old Thoroughbred horses received single calcein injections 19 and 8 days prior to euthanasia, labeling sites of active mineralization with fluorescent bands. Confocal scanning laser microscopy images of articular calcified cartilage (ACC) from distal third metacarpal condyles were registered to qBSE images of the same sites using an in-house program. ImageJ and Sync Windows enabled the simultaneous collection of LAR and Dm data. The repeatability of the registration and measurement protocols was determined. Dm profiles between calcein labels were explored for an association with time. Dm was 119.7 +/- 24.5 (mean +/- standard deviation) gray levels (where 0 = backscattering from monobrominated and 255 from monoiodinated dimethacrylate standards, respectively), while modal and maximum LAR were 0.45 and 3.45 microm/day, respectively. Coefficients of variation (CV) for Dm were 0.70 and 0.77% with and without repeat registration, respectively; CVs for LAR were 1.90 and 2.26% with and without repeat registration, respectively. No relationship was identified between Dm and time in the 11-day interlabel interval. Registration of CSLM to qBSE images is sufficiently repeatable for quantitative studies of equine ACC.     

Serological characterization and prevalence of spvR genes in Salmonella isolated from foods involved in outbreaks in Brazil

Salmonella strains (n = 75) isolated from foods involved in foodborne outbreaks occurred in Rio Grande do Sul State, Brazil, during 1999 and 2000 were studied. Strains were serotyped and submitted to PCR analysis to verify the prevalence of Salmonella plasmid virulence (spvR) regulatory gene. Among the 75 isolates, 73 (97%) were classified as Salmonella enterica serovar Enteritidis. All of the Salmonella strains isolated in 1999 were classified as serotype Enteritidis, whereas in 2000 two isolates were serotyped as Salmonella Derby and Salmonella Typhimurium. Regarding the prevalence of spvR gene, 62 strains (82.7%) were PCR positive, and a positive correlation (P < 0.05) between the strains of Salmonella Enteritidis and the presence of spvR gene was demonstrated, which suggests that this gene is a characteristic of the Salmonella Enteritidis analyzed.     

Effect of protein level in prepartum diets on metabolism and performance of dairy cows

Multigravid Holstein cows (n = 75) were used in a randomized block design to evaluate the effect of prepartum diets formulated to supply surplus energy and incremental concentrations of protein on the nutritional status of dairy cows at parturition. Cows were blocked according to expected calving date and assigned to one of five diets: 9.7, 11.7, 13.7, 14.7, and 16.2% crude protein (CP). Dietary treatments were initiated 28 d before expected calving date and fed until parturition. A common diet was fed postpartum. Dry matter intake and milk yield were recorded daily through 90 d postpartum. Increasing the protein concentration from 9.7 to 14.7% of dry matter during the last 28 d of gestation improved responses of cows during lactation. Increasing dietary protein up to 14.7% also increased milk yield response to recombinant bovine somatotropin (rbST) during the ninth week of lactation and yields of 305-d 2x mature equivalent milk, milk protein, and milk fat. Plasma aspartate aminotransferase tended to be highest in cows fed 13.7 and 14.7% CP prepartum, but decreased linearly postpartum in response to dietary protein levels. There were no treatment differences for plasma insulin-like growth factor-1 (IGF-1) at d 60 postpartum (before rbST provision), but IGF-1 on d 90 (after rbST provision) was higher in plasma of cows fed 14.7% CP than the other diets except 13.7% CP. Close-up diets containing 13.7% CP and surplus energy produced the most beneficial outcomes during the subsequent lactation.     

Diminished macrophage cholesterol removal rate by the altered HDL metabolism in the Nagase analbuminemic rat

Dyslipoproteinemia of the Nagase analbuminemic rat (NAR) is characterized by elevated concentrations of VLDL and LDL attributed to increased rates of liver lipoprotein synthesis. Increased lysophosphatidylcholine (LPC) in NAR HDL has been attributed to high plasma LCAT activity. We show here that, as compared with Sprague-Dawley rats (SDR), NAR plasma triacylglycerol (TAG), total cholesterol (TC), HDL TAG, protein, total phospholipids (PL), LPC, and PS are increased. These alterations rendered the NAR HDL particle more susceptible to the activity of the enzyme hepatic lipoprotein lipase (HL), which otherwise was unaltered in our study. Fractional catabolic rates in blood of the autologous ¹²⁵I-apoHDL (median and lower quartile values), were, respectively, 0.231 and 1.645 (n=10) in NAR as compared with 0.140 and 0.109 (n=10) in SDR (P=0.012), corresponding to synthesis rates of HDL protein of 89.8±33.7 mg/d in NAR and 17.4±6.5 mg/d in SDR (P=0.0122). Furthermore, Swiss mouse macrophage free-cholesterol (FC) efflux rates, measured as the percent [¹⁴C]-cholesterol efflux/6 h, were 8.2±2.3 (n=9) in NAR HDL and 11.2±3.2 (n=10) in SDR HDL (P=0.03). Therefore, in NAR the modification of the HDL composition slows down the cell FC efflux rate, and together with the increased rate of plasma HDL metabolism influences the reverse cholesterol transport system.     

Oxidation of H2S by iron oxides in unsaturated conditions

Previous studies have demonstrated that gas-phase H2S can immobilize certain redox-sensitive contaminants (e.g., Cr, U, Tc) in vadose zone environments. A key issue for effective and efficient delivery of H2S in these environments is the reactivity of the gas with indigenous iron oxides. To elucidate the factors that control the transport of H2S in the vadose zone, laboratory column experiments were conducted to identify reaction mechanisms and measure rates of H2S oxidation by iron oxide-coated sands using several carrier gas compositions (N2, air, and O2) and flow rates. Most experiments were conducted using ferrihydrite-coated sand. Additional studies were conducted with goethite- and hematite-coated sand and a natural sediment. Selective extractions were conducted at the end of each column experiment to determine the mass balance of the reaction products. XPS was used to confirm the presence of the reaction products. For column experiments in which ferrihydrite-coated sand was the substrate and N2 was the carrier gas, the major H2S oxidation products were FeS and elemental sulfur (mostly S8(0), represented as S(0) for simplicity) at ratios that were consistent with the stoichiometry of the postulated reactions. When air or O2 were used as the carrier gas, S(0) became the dominant reaction product along with FeS2 and smaller amounts of FeS, sulfate, and thiosulfate. A mathematical model of reactive transport was used to test the hypothesis that S(0) forming on the iron oxide surfaces reduces access of H2S to the reactive surface. Several conceptual models were assessed in the context of the postulated reactions with the final model based on a linear surface poisoning model and fitted reaction rates. These results indicate that carrier gas selection is a critical consideration with significant tradeoffs for remediation objectives.     

Cholesterol and fatty acids profile of Brazilian commercial chicken giblets

This study was carried out to determine the chemical composition, cholesterol contents and fatty acids profile of Brazilian commercial chicken giblets. The analysis were performed in gizzard, liver and heart in natura and also in cooked gizzard, fried liver and roasted heart. Fat and cholesterol contents ranged from 0.88% and 72.68 mg/100 g, in cooked gizzard, to 22.19% and 213.18 mg/100 g, in roasted heart. As the fat content gets higher, so does the cholesterol content. Palmitic (C16:0) and stearic acids (C18:0) were the predominant saturated fatty acids (SFA). The C16:0 ranged from 6.39% in cooked gizzard to 18.51% in fried liver. The C18:0 level ranged from 6.62% in roasted heart to 19.19% in cooked gizzard. Linoleic acid (C18:2 omega 6) was the predominant polyunsaturated fatty acid (PUFA). The data revealed that the three different analysed giblets presented a good PUFA/SFA ratio, with values of 1.11, 1.14 and 1.40 for cooked gizzard, fried liver and roasted heart, respectively.     

Growth of Listeria monocytogenes in different retail delicatessen meats during simulated home storage

Delicatessen meats are reported to be the leading vehicle of foodborne listeriosis in the United States. Listeria monocytogenes can reach high numbers in these products during storage, and the growth rate is largely dictated by product formulation and storage temperature. To assess the impact of product age on Listeria growth, five commercial brands each of cured and uncured turkey breast, ham, and roast beef (three lots per brand) were sliced (approximately 25 g per slice) at the beginning of the shelf life, the midpoint, and the last allowable day of sale, surface inoculated with an eight-strain cocktail of L. monocytogenes (approximately 40 CFU/g), and then quantitatively examined for Listeria, lactic acid bacteria, and mesophilic aerobic bacteria during aerobic storage at 4, 7, or 10°C. As expected, L. monocytogenes grew faster in deli meats without rather than with Listeria inhibitors (lactate and/or diacetate) and at the highest storage temperature (10°C). Lag-phase durations for L. monocytogenes in deli meats with and without Listeria inhibitors were 9.21, 6.96, and 5.00 and 6.35, 3.30, and 2.19 days at 4, 7, and 10°C, respectively. Generation times for L. monocytogenes in deli meats with and without Listeria inhibitors were 1.59, 1.53, and 0.85 and 0.94, 0.50, and 0.36 at 4, 7, and 10°C, respectively. Maximum population densities for L. monocytogenes in deli meats with and without Listeria inhibitors were 5.26, 5.92, and 5.97 and 8.47, 8.96 and 9.34 log CFU/g at 4, 7, and 10°C, respectively. Although lactate and diacetate suppressed L. monocytogenes growth, the extent of inhibition differed, ranging from total inhibition in roast beef to only partial inhibition in ham and cured turkey. Listeria growth was also impacted by lot-to-lot variation in the concentrations of Listeria inhibitors, product pH, and background microflora. These data will be useful for developing recommendations for "best consumed by" dating for deli meats using a risk-based approach.     

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