Optimization of a Staphylococcus aureus adhesion assay for equine corneocytes

Meticillin-resistant Staphylococcus aureus (MRSA) causes serious skin and soft-tissue infections of humans and animals. Multiple strains of MRSA have been characterized, and one in particular, designated as strain USA 500, causes infections predominantly of horses and the people who work with them. The purpose of this study was to optimize an assay which could subsequently be used to compare the relative avidity of different S. aureus strains for equine corneocytes. Corneocytes were collected from the perineal skin of 10 healthy horses onto adhesive discs. The discs were then incubated at 37°C with an S. aureus field strain at each of three concentrations [10(7), 10(8) and 10(9) colony forming units (CFU)/mL] and for each of three incubation periods (45, 90 and 180 min). After standardized rinsing and staining procedures, discs were examined at ×1,000 magnification and areas containing confluent corneocytes photographed. The percentage of surface area occupied by adherent bacteria was analysed using image processing and analysis software. Significant colour space image processing was required to distinguish bacteria from the ubiquitous melanin granules present within equine corneocytes. Objective and subjective methods were used to determine optimal conditions for specific adherence without introducing confounding factors. A bacterial concentration of 10(8) CFU/mL incubated with corneocytes for 45 min produced maximal bacterial adhesion with the least amount of interbacterial clumping. Future studies should utilize these conditions for optimal assay performance.     

Optimization of lycopene extraction from tomato cell suspension culture by response surface methodology

Radioisotope-labeled lycopene is an important tool for biomedical research but currently is not commercially available. A tomato cell suspension culture system for the production of radioisotope-labeled lycopene was previously developed in our laboratory. In the current study, the goal was to optimize the lycopene extraction efficiency from tomato cell cultures for preparatory high-performance liquid chromatography (HPLC) separation. We employed response surface methodology (RSM), which combines fractional factorial design and a second-degree polynomial model. Tomato cells were homogenized with ethanol, saponified by KOH, and extracted with hexane, and the lycopene content was analyzed by HPLC-PDA. We varied five factors at five levels: ethanol volume (1.33-4 mL/g); homogenization period (0-40 s/g); saturated KOH solution volume (0-0.67 mL/g); hexane volume (1.67-3 mL/g); and vortex period (5-25 s/g). Ridge analysis by SAS suggested that the optimal extraction procedure to extract 1 g of tomato cells was at 1.56 mL of ethanol, 28 s homogenization, 0.29 mL of KOH, 2.49 mL of hexane, and 17.5 s vortex. These optimal conditions predicted by RSM were confirmed to enhance lycopene yield from standardized tomato cell cultures by more than 3-fold.     

Response of <Emphasis Type="Italic">Pinus pseudostrobus</Emphasis> Lindl. to fertile growing medium and tephra-layer depth under greenhouse conditions

Major disturbances that remove vegetation cover create conditions that differ greatly from those in adjacent forests. In volcanic areas, tephra deposition can be a major disturbance that eliminates vegetation cover and creates a barrier for plant establishment. The eruption of the Paricutín volcano, in Michoacán, México, from 1943 to 1952, created large tephra deposits adjacent to forest stands. Many of these deposits still lack native vegetation and understanding of the limiting factors for establishment of native tree species is necessary to restore these areas. Pinus pseudostrobus early growth in response to the amount of fertile growing medium on top of a variable-depth tephra layer was evaluated in a greenhouse experiment. Increasing amounts of fertile growing medium increased height of 28-week-old plants (no growing medium = 5.5 cm tall ±1.8 cm, 3 cm of growing medium = 21.2 ± 4.6 cm and 6 cm of growing medium 24.8 ± 4.4 cm). Other variables showed the same trend (diameter, number of fascicles, belowground biomass and aboveground biomass). Our results suggest that P. pseudostrobus establishment in tephra covered areas will depend on the accumulation of a fertile layer of more than 3 cm in thickness.     

Nutrient cycling in two traditional Central American agroforestry systems

A preliminary nutrient cycling study quantified total and temporal nutrient inputs via litterfall and pruning residues in two agroforestry systems: (1) Coffea arabica (perennial crop)-Erythrina poeppigiana (leguminous shade tree); and (2) C. arabica-E. poeppigiana-Cordia alliodora with emphasis on the effect of the timber tree C. alliodora. The total annual input of litterfall plus pruning residues was similar in both associations. Total annual input from E. poeppigiana was less than half in the association with C. alliodora than without, but the litterfall from this latter species compensated for the loss. Large differences in the total annual nutrient input of K, Ca and Mg was found between associations, but not for N or P. The amount of nutrients recycled by the associated trees reached the recommended level of fertilizer required for coffee production. The inclusion of C. alliodora within the C. arabica-E. poeppigiana association resulted in a more evenly distributed annual nutrient input.     

Comparison of the effects of buprenorphine,oxymorphone hydrochloride,and ketoprofen for postoperative analgesia after onychectomy or onychectomy and sterilization in cats

In this prospective, randomized, blinded study, 68 clinically healthy cats that had onychectomy (n = 20), onychectomy and castration (n = 20), or onychectomy and ovariohysterectomy (n = 28) were randomly assigned to one of four postoperative analgesic treatment groups: buprenorphine (0.01 mg/kg body weight, intramuscularly [IM]), oxymorphone hydrochloride (0.05 mg/kg body weight, IM), ketoprofen (2 mg/kg body weight, IM), and placebo (physiological saline). Sedation scores, visual analog pain scores, cumulative pain scores, serum cortisol concentration, and appetite were used to assess postoperative analgesic effect. Buprenorphine demonstrated the highest efficacy with the lowest cumulative pain scores and serum cortisol levels.     

Effects of Monoglycerides of Varying Fatty Acid Chain Length and Mixtures Thereof on Sponge‐and‐Dough Breadmaking Quality

Monoglycerides are widely used in the baking industry because of their antistaling effects, mainly suppressing crumb firming. Commercial monoglycerides are normally prepared from hydrogenated fats, with stearate being the most common fatty acid. In a previous study, monoglycerides such as monopalmitate (C16) and monostearate (C18) had positive effects on Canadian short process bread but no improvements on sponge‐and‐dough process (SDP) bread. The objective of this study was to investigate the effects of saturated monoglycerides of varying fatty acid chain length (C14–C22) on SDP breadmaking quality by using volume judgment, crumb image analysis, and texture measurements. Higher levels (1.00–1.50%) of all monoglycerides (C14, C16, and C18) significantly (P < 0.05) increased loaf volume and cell diameter. The larger cell diameter with increasing levels of these monoglycerides may have resulted from softer, more extensible dough handling properties and greater gas cell stability during baking. Addition of C16 and C18 caused the largest increase in crumb softness with increasing monoglyceride levels but showed relatively low resilience, which might be related to larger loaf volume (i.e., lower density of bread). However, addition of blended monoglycerides C14+C16 increased crumb softness and loaf volume while partially retaining resilience. Each monoglyceride had a different function in breadmaking quality and somewhat positive effects on SDP.