A rapid molecular method for diagnosing epidemic dermatophytosis in a racehorse facility.

REASONS FOR PERFORMING STUDY: Identification of the species and strain of dermatophyte can play an effective role in control of disease outbreaks by establishing the source of infection. Current methods of identification are based on cultural and microscopic methods, often involving weeks before a positive identification are made. A rapid molecular diagnostic method would therefore be an important laboratory technique, but requires confirmation in equine clinical practice. OBJECTIVES: To test the sensitivity and specificity of molecular diagnostic methods applied to a racehorse herd from the Korean Racehorse Authority (KRA). METHODS: A total of 57 DNA samples were collected from hairs and crusts of skin lesions in KRA racehorses with histories and clinical signs suggestive of dermatophytosis, which was confirmed by dermatophyte-specific PCR amplification analysis using the primer pair for the chitin synthase 1 (CHS1) gene. RESULTS: Thirty-eight racehorses were definitively diagnosed with dermatophytosis using molecular and traditional diagnostic methods. PCR fingerprinting profiles using simple repetitive (GACA)4 primers showed that all diagnosed horses had the same pattern profile. Oligonucleotide sequencing of CHS1 gene PCR products confirmed Trichophyton mentagrophytes as the infectious agent. CONCLUSIONS: This study demonstrates that the PCR-based molecular diagnostic method is sensitive and specific and offers fast precise diagnosis of dermatophytosis in horses.

Properties of duck meat sausages supplemented with cereal flours.

Duck meat sausages were prepared using 10% beef fat (FDS) and 10% hydrated cereal flours including rice (RDS), wheat, corn, millet, and barley to replace fat. Control duck sausages (DS) were also prepared only with duck meat and duck meat plus 10% beef fat. Results showed that protein and fat contents significantly decreased and total expressible fluid reduced with the addition of cereal flours in duck sausage batters. The FDS had higher fat content and lower pH compared with others. Duck sausages with 10% supplemented wheat flour showed the lowest cooking loss among sausages and had similar redness and chroma values to FDS and DS. Texture analysis indicated that hardness of duck sausage significantly decreased when cereal flours and beef fat were added. In particular, RDS showed the lowest values for all texture measurements compared with others. Result of moisture absorption capacity suggested that the decrease in hardness in RDS was due to higher moisture retention for rice flour treatment. Sensory evaluation indicated that DS had significantly lower overall acceptability than RDS, due to its off-flavor, whereas RDS had higher overall acceptability than DS.

Discoloration characteristics of 3 major muscles from cattle during cold storage.

Discoloration characteristics of 3 major muscles (LD, Longissimus dorsi; PM, Psoas major; SM, Semimemebranosus) from Korean native cattle (Hanwoo) were monitored during 7 d of cold storage at 4 degrees C. The muscles were obtained from 12 Hanwoo carcasses at 24 h postmortem. Meat color (CIE L*, a*, b*), myoglobin (Mb) concentration, chemical form, metmyoglobin (MetMb) reducing ability (MRA), mitochondria concentration, and thiobarbituric acid reactive substance (TBARS) were measured at 1, 3, 5, and 7 d of storage. Although there were no significant differences in CIE a* and b*-values between the 3 muscles at day 1, the values of PM muscle were significantly (P < 0.05) lower than those of LD and SM muscles at day 5 and 7. PM muscle showed a rapid decrease in the oxymyoglobin (OxyMb) and an increase in MetMb, which resulted in a significantly (P < 0.05) higher percentage of MetMb in PM muscle compared to LD and SM muscles. Also, the Mb and mitochondria concentration of PM muscle was significantly (P < 0.05) higher than those of LD and SM muscles. However, there were no significant differences in MRA, pH, or TBARS between the 3 muscles during 7 d of cold storage. It was concluded that rapid discoloration (that is, MetMb accumulation) in PM muscle of Hanwoo could be due to its higher contents of Mb and mitochondria.

Effects of various fiber additions on lipid digestion during in vitro digestion of beef patties.

The purpose of this study was to examine the effect of various fiber additions on lipid digestion during the in vitro digestion of beef patties. The control patties were prepared with 90.5% lean meat and 9.5% tallow. Treatments consisted of 90% lean meat with 9.5% tallow and either 0.5% cellulose, 0.5% chitosan, or 0.5% pectin. The beef patties were then passed through an in vitro digestion model that simulated the composition of the mouth, stomach, and small intestine juices. The change in structure and properties of the lipid droplets was monitored by laser scanning confocal fluorescence microscopy. In general, there was a decrease in lipid droplet diameter as the droplets moved from mouth to stomach to small intestine. The amount of free fatty acid dramatically increased after in vitro digestion in all beef patties. The amount of free fatty acid was, however, lower in beef patties containing chitosan and pectin than other beef patties after in vitro digestion. Beef patties containing various fibers had lower thiobarbituric acid-reactive substances (TBARS) values than samples with no fibers. Among the samples to which fibers were added, chitosan and pectin had lower TBARS than beef patties with cellulose. The cholesterol content decreased after in vitro digestion in all beef patties but was not different among the beef patties before and after in vitro digestion. These results enhance our understanding of the physicochemical and structural changes that occur to ground beef within the gastrointestinal tract.

Effect of chilling temperature of carcass on breast meat quality of duck.

An experiment was carried out to investigate whether variations in chill water temperature affect muscle shortening and meat quality in duck breast. Three chill water temperatures were applied to duck carcasses at 20 min postmortem for 30 min, including in ice water at 0 degrees C, in cold water at 10 degrees C, and in water at 20 degrees C. Results revealed that carcass temperatures were different (P < 0.05) at 50 and 120 min of postmortem with lower temperatures at the 0 degrees C treatment (P < 0.05). The pH over the first 24 h postmortem was not different (P > 0.05) among treatments, with the exception of 50 min postmortem. The pH of breast meat in the 0 degrees C treatment was higher (P < 0.05) than that of 20 degrees C treatment at 50 min postmortem (just after chilling). No other differences (P > 0.05) in pH existed among treatments. Drip loss, cooking loss, and moisture content were not different for breast meat samples that were chilled at different temperatures. Differences (P < 0.05) were found in CIE (L, a, and b) color values. Lightness (L) increased, whereas redness (a) decreased as the chill water temperature increased. Lower yellowness (b) was found in the breast meat samples at the 10 degrees C chill water temperature. However, shear force, sarcomere length, and protein solubility were not different (P > 0.05) among the breast meat samples chilled at different chill water temperatures. It may be concluded that chilling duck carcasses at different temperature ranges from 0 to 20 degrees C did not influence muscle shortening or meat quality, except in regard to breast meat color.

The relationship of physiological maturity and marbling judged according to Korean grading system to meat quality traits of Hanwoo beef females.

Fifty seven carcasses from Hanwoo beef females were randomly selected by official meat graders and were sorted into three levels of maturity and marbling. Carcass data was collected for back fat thickness, longissimus area, carcass weight, meat colour, fat colour, marbling score, yield and quality grades. Mature carcasses had more yellow fat, coarser texture, a larger longissimus muscle area and lower quality grades and marbling scores (P<0.05). Carcasses with a higher marbling score had thicker fat and a higher quality grade. Carcasses with low marbling had a higher yield grade and a coarser texture (P<0.05). Higher marbling scores corresponded with lower cook and drip loss values for longissimus steaks. As the maturity of carcass was increased, the redness and lightness of meat and the yellowness of fat all tended to increase. Tenderness, flavour and overall acceptability scores for the older maturity group were lower than for younger and intermediate groups. Marbling was significantly (P<0.01) correlated with quality grade, crude fat content, cook and drip losses, and Warner-Bratzler shear force. The maturity level was also significantly (P<0.01) correlated with quality grade, fat colour, texture score, number of calves produced and milk teeth, meat redness and yellowness, fat yellowness, and Warner-Bratzler shear force. Results indicate that a low marbling group and older maturity group based on Korean grading system could negatively influence carcass traits and beef qualities of Hanwoo beef female.

Effects of conjugated linoleic acid on color and lipid oxidation of beef patties during cold storage.

The effects of conjugated linoleic acid (CLA) on color and lipid oxidation of beef patties were investigated. Ground beef was divided into three batches. The control patties were prepared with 90% lean meat and 10% tallow. The second treatment consisted of 90% lean meat with 9.5% tallow+0.5% CLA sources. The third treatment consisted of 90% lean meat with 8% tallow+2% CLA sources. The patties were wrap-packaged and then stored at 4° for 14 days. The CLA concentration significantly increased (P<0.05) by substituting CLA sources for fat. Storage of the patties did not alter the CLA concentration in beef patties. The treatment substituted with CLA sources had significantly lower TBARS (2-thiobarbituric acid-reactive substances) values (P<0.05) than the control. For oxymyoglobin contents and a* value, substituted CLA sources treatments had significantly higher values than the control. However, L* value significantly increased by substituting CLA sources for fat.

Effects of dietary conjugated linoleic acid on fatty acid composition, lipid oxidation, color, and water-holding capacity of pork loin.

The effects of dietary conjugated linoleic acid (CLA) on fatty acid composition, lipid oxidation, and pork quality were investigated. Pigs (n = 20) were fed a diet containing 0, 1, 2.5, or 5% CLA for 4 wk and slaughtered at 105 kg. The longissimus thoracis et lumborum muscle was collected at 24 h postmortem. Pork loin chops (3 cm thick) were packaged aerobically and stored at 4 degrees C for 7 d. Samples were analyzed for ultimate pH, intramuscular fat content, fatty acid composition, thiobarbituric acid-reactive substances, color (L*, a*, b*), and water-holding capacity. Dietary CLA reduced the concentration of linoleic acid and increased CLA concentration in intramuscular fat of pork loin (P < 0.05). The concentration of CLA in muscle was increased with dietary CLA level and did not change during storage. Thiobarbituric acid-reactive substance value of control was higher than that of the CLA-fed groups (P < 0.05). Intramuscular fat content was increased by dietary CLA, and less purge loss was observed with samples from CLA-fed pigs (P < 0.05). Dietary CLA improved the color stability of pork loin during cold storage. After 7 d, lightness (L*) and yellowness (b*) of the 5% CLA-fed group were significantly lower than those of control (P < 0.05). The results indicated that the water-holding capacity of pork loin was increased with increased intramuscular fat content apparently caused by dietary CLA. Also, the data indicated that color stability of pork was improved with inhibition of lipid oxidation and changing of fatty acid composition by dietary CLA.

Influence of slaughter weight and sex on yield and quality grades of Hanwoo (Korean native cattle) carcasses.

To assess the effects of slaughter weight and sex on APGS (Animal Products Grading Service) quality and APGS yield grade of Korean Hanwoo (n = 20,881) cattle, data were collected from cow, bull, and steer carcasses during a 1-yr period. Factors used to determine quality grade (marbling, meat color, fat color, texture, and overall maturity score) and yield grade (cold carcass weight, adjusted fat thickness, and longissimus muscle area) by the Korean grading system were recorded. Both yield and quality grades were improved (P < 0.01) with heavier slaughter weight, but there was no difference in yield grade for Hanwoo cattle classes heavier than 551 kg (P > 0.01). Longissimus muscle area, adjusted fat thickness, and marbling score increased (P < 0.01) with carcass weight. Bull carcasses showed higher yield but lower quality than those of cows or steers (P < 0.01). The quality grade of steer carcasses was higher (P < 0.01) than that of cow carcasses due to higher marbling scores, lower maturity scores, and heavier carcass weights. Hanwoo carcasses with larger longissimus muscle areas in relation to their carcass weight had lower APGS quality grades. The APGS quality grades were different between yield grade A and B carcasses (P < 0.01), but quality grade was not improved by increased fat thickness beyond the point of yield grade B. Adjusted fat thickness and marbling score showed significant (P < 0.01) differences among all yield grade classes, and this resulted in increased quality grade as yield grade decreased. Adjusted fat thickness showed the strongest correlation (r = -0.63) with yield grade, whereas marbling score had the strongest correlation (r = 0.81) with quality grade. Results showed a negative effect of castration on yield but a positive effect on quality. Also, data showed that Hanwoo carcasses with heavier weights had higher quality grades than those of lighter weight.

Improvement of oxidative stability of conjugated linoleic acid (CLA) by microencapsulation in cyclodextrins.

Oxidative stability of conjugated linoleic acid (CLA) encapsulated in alpha-, beta-, and gamma-cyclodextrins (designated CLA/CDs microencapsules) was studied by measuring the headspace-oxygen depletion in airtight serum bottles and by measuring the peroxide values (POV). The rate of oxygen depletion was reduced from 41.0 (control) to 21.5, 2.1, 1.2, and 1.1 micromol/L.h(-)(1) by CLA/alpha-CD microencapsules at 1:1, 1:2, 1:4, and 1:6 mole ratios, respectively, indicating that CLA oxidation was completely protected by a 1:4 mole ratio of CLA/alpha-CD. Such a protective effect by CLA/beta-CD or CLA/gamma-CD microencapsules was achieved at a 1:6 mole ratio, but the effect by CLA/beta-CD was slightly greater than that by CLA/gamma-CD. The protective effect of alpha-, beta-, and gamma-CDs for CLA oxidation was confirmed by their POV-reducing abilities in CLA/CDs. These results suggest that alpha-CD was the most effective for the protection of CLA oxidation by microencapsulation, followed by beta-CD and gamma-CD.

The relationship of sarcoplasmic and myofibrillar protein solubility to colour and water-holding capacity in porcine longissimus muscle.

In order to investigate the relationship of sarcoplasmic and myofibrillar protein solubility to colour and water-holding capacity (WHC) in pork, 60 loins were selected to represent the quality classes: PSE (pale, soft, exudative), RSE (reddish-pink, soft, exudative), RFN (reddish-pink, firm, non-exudative) and DFD (dark, firm, dry). PSE samples exhibited lower (p<0.05) protein solubility (sarcoplasmic, myofibrillar and total) compared to the other quality classes. RSE samples exhibited lower (p<0.05) sarcoplasmic protein solubility compared to DFD samples. RSE, RFN and DFD samples had similar myofibrillar and total protein solubilities. Sarcoplasmic protein solubility explained 71% of the variation in lightness with a linear decrease in L* value. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) gels of the sarcoplasmic and myofibrillar samples distinctly showed the association of some sarcoplasmic proteins with the myofibrillar protein fractions in PSE and RSE samples. The sarcoplasmic proteins which precipitated were phosphorylase, creatine kinase, triose phosphate isomerase and myokinase for PSE and phosphorylase for RSE samples. Pork colour is highly correlated with precipitation of sarcoplasmic proteins while WHC is affected by denaturation of myofibrillar proteins (PSE samples) and lower ultimate pH (PSE and RSE samples).

The physicochemical properties, plasma enzymatic hydrolysis, and nasal absorption of acyclovir and its 2'-ester prodrugs.

A series of 2'-(O-acyl) derivatives of 9-(2-hydroxyethoxymethyl)guanine (acyclovir) was synthesized by acid anhydride esterification. Aqueous solubilities in isotonic phosphate buffer (pH 7.4), partition coefficients in 1-octanol/phosphate buffer, and hydrolysis kinetics in rat plasma were determined. The ester prodrugs showed consistent increases in lipophilicity with corresponding decreases in aqueous solubility as a function of side-chain length. The bioconversion kinetics of the prodrugs appear to depend on both the apolar and the steric nature of the acyl substituents. When perfused through the rat nasal cavity using the in situ perfusion technique, acyclovir showed no measurable loss from the perfusate. Nasal uptake of acyclovir prodrugs, on the other hand, were moderately improved. Furthermore, the extent of nasal absorption appears to depend on the lipophilicity of the prodrugs in the descending order hexanoate > valerate > pivalate > butyrate. Simultaneous prodrug cleavage by nasal carboxylesterase was also noted in the case of hexanoate.

Acyclovir permeation enhancement across intestinal and nasal mucosae by bile salt-acylcarnitine mixed micelles.

The purpose of this study was to investigate the absorption enhancement of acyclovir, an antiviral agent, by means of bile salt-acylcarnitine mixed micelles. The specificity, site dependence, palmitoyl-DL-carnitine chloride (PCC) concentration dependence, and effects of absorption promoters on acyclovir absorption via the nasal cavity (N) and four different intestinal segments of the rat, i.e., duodenum (D), upper jejunum (UJ), combined lower jejunum and ileum (LJ), and colon (C) were evaluated. The present study employed the rat in situ nasal and intestinal perfusion techniques and utilized sodium glycocholate (NaGC), three acylcarnitines, and their mixed micelles as potential nasal and intestinal absorption promoters. Acylcarnitines used were DL-octanoylcarnitine chloride (OCC), palmitoyl-DL-carnitine chloride (PCC), and DL-stearoylcarnitine chloride (SCC). All acylcarnitines and NaGC by themselves produced negligible enhancement of acyclovir absorption in the rat intestine, while OCC and SCC were totally ineffective in the nasal cavity. However, the mixed micellar solutions of NaGC with PCC or SCC could significantly increase the mucosal membrane permeability of acyclovir in the colon and nasal cavity. On the other hand, NaGC-OCC mixed micelles slightly increased the absorption of acyclovir by both routes. When a mixed micellar solution of NaGC with PCC was used, the rank order of apparent acyclovir permeability (Papp; cm/sec), corrected for surface area of absorption, was N (10.54 +/- 0.62 x 10(-5)) > D (6.82 +/- 0.30 x 10(-5)) > LJ (2.90 +/- 0.08 x 10(-5)) > C (2.54 +/- 0.14 x 10(-5)) > UJ (2.30 +/- 0.22 x 10(-5)).(ABSTRACT TRUNCATED AT 250 WORDS)

Mechanism and site dependency of intestinal mucosal transport and metabolism of thymidine analogues.

This study has been undertaken to investigate the mechanisms of intestinal mucosal transport and metabolism of thymidine analogues and to identify any optimal site(s) of the rat intestine particularly involved in the absorption of thymidine analogues. The intestinal absorption of 3'-azido-3'-deoxythymidine (AZT) was studied at three initial concentrations in four segments of the rat intestine using an in situ recirculating perfusion technique. Disappearance of AZT followed first-order kinetics throughout the gastrointestinal (GI) tract at all tested concentrations. The apparent first-order rate constants were found to be relatively invariant over a broad range of concentrations from 0.01 to 1.0 mM. Corrected for the length of each segment, the apparent permeability (Papp) of AZT was 3.01 +/- 0.32 x 10(-5) cm/sec (mean +/- SE) in the duodenum, 2.06 +/- 0.24 x 10(-5) cm/sec in the upper jejunum, 0.76 +/- 0.13 x 10(-5) cm/sec in the combined lower jejunum and ileum, and 0.32 +/- 0.10 x 10(-5) cm/sec in the colon, which indicated that intrinsic absorptivity was greater in the upper GI tract than in the lower portions possibly due to the differences in surface area for absorption. No AZT metabolite appeared in any part of the GI tract. On the other hand, thymidine and other analogues, i.e., 5-iodo-2'-deoxyuridine and 2'-deoxyuridine, were rapidly metabolized into nucleobase and sugar in the upper GI tract, whereas in the colon no metabolite appeared. A free 3'-OH group appears to be necessary for the metabolism (catabolism) of thymidine analogues in the rat intestine mainly by pyrimidine nucleoside phosphorylase.(ABSTRACT TRUNCATED AT 250 WORDS)

Relative bioavailability and pharmacokinetics: a combination of pentazocine and acetaminophen.

The relative bioavailability and pharmacokinetics of a combination product containing pentazocine and acetaminophen were studied in 20 healthy human males. Each subject, in a single-dose three-way crossover design, received two different preparations containing 50 mg of pentazocine (as base) and 1300 mg of acetaminophen either as capsule-shaped tablets or as a solution. Plasma concentrations of pentazocine and acetaminophen were determined from 0.25 to 12 h following oral administration. The plasma data for both compounds in the tablet formulation were described by an open one-compartment body model with first-order absorption. The average (+/- SD) bioavailability of the tablet relative to that of the solution was 85.0 +/- 31.1 and 88.6 +/- 13.1% for pentazocine and acetaminophen, respectively. The apparent first-order regression-dependent elimination rate constants for pentazocine from the tablet and solution preparations were 0.19 +/- 0.08 and 0.20 +/- 0.06 h-1, respectively, while the rate constants for acetaminophen were 0.26 +/- 0.03 and 0.25 +/- 0.03 h-1 for the tablet and solution preparations, respectively. These rate constants correspond to terminal elimination half-lives of approximately 3.6 h for pentazocine and approximately 2.7 h for acetaminophen.

Dose proportionality of amrinone.

Amrinone was given to 18 healthy subjects in doses of 75, 150, and 225 mg in a randomized crossover design. Plasma levels were shown to rise in proportion to dose. The mean plasma AUC, extrapolated to infinite time, was determined for each dose level; the values obtained were 4, 8.18, and 12.35 micrograms . hr/ml for the 75-, 150-, and 225-mg doses. Mean maximum observed plasma concentrations were 1.03, 1.74, and 2.58 micrograms/ml. At higher doses the extrapolated AUC is more variable, but it is linear over the range of 0.73 to 3.81 mg/kg. The apparent first-order terminal elimination rate is not dose dependent and corresponds to a t1/2 of 3.85 hr.

Oral bioavailability and intravenous pharmacokinetics of amrinone in humans.

Fourteen healthy males received two 75-mg doses of amrinone as a single capsule and as an intravenous solution in a single-dose crossover study. The mean (+/-SD) bioavailability, based on the area under the plasma concentration versus time curves, was 0.93 +/- 0.12. The plasma data for these subjects during the intravenous phase was described by an open two-compartment body model with a mean (+/-SD) apparent first-order terminal elimination rate constant, beta, of 0.19 +/- 0.06 hr-1, which corresponds to a half-life of 3.6 hr.

The pharmacokinetics and pharmacodynamics of sulfinalol hydrochloride in dogs after intravenous administration.

The pharmacokinetic behavior of sulfinalol hydrochloride, an antihypertensive agent with vasodilator and beta-adrenergic blocking activity, was determined in dogs after intravenous administration. The plasma concentrations of sulfinalol HCl were fit to an open two-compartment body model. The mean values for the alpha- and beta-phase constants were 33.3 hr-1 and 0.52 hr-1, respectively. The mean plasma clearance was 2.30 L/kg X hr. The steady-state volume of distribution was approximately four times the body weight of the animals. The urine data gave renal clearance rates approximately equal to the normal glomerular filtration rate in the dog. About 7.5% of the administered dose was excreted in the urine as free sulfinalol hydrochloride. The time course of the hypotensive effect of sulfinalol appears to be better correlated with calculated tissue levels of drug than with observed plasma levels.