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Biomedical subjects

A J Yee

Publications and source records attributed to A J Yee.

At least 19 recordsLinked to original sources

Calibration and validation of an air-displacement plethysmography method for estimating percentage body fat in an elderly population: a comparison among compartmental models.

BACKGROUND: The use of hydrostatic weighing (HW) to measure body composition in the elderly can be difficult and is based on the assumption of constancy of body compartments. OBJECTIVE: We calibrated and validated a new air-displacement plethysmography (AP) method for measuring body composition in the elderly. DESIGN: A 4-compartment equation for calculating percentage body fat (%BF) that used body density (D(b)), total body water, and bone mineral content was used as the criterion for evaluating %BF estimated by the 2- and 3-compartment models. D(b) was measured by HW [D(b(HW))] and by use of the AP instrument [D(b(AP))] in 30 elderly men and 28 elderly women aged 70-79 y. RESULTS: D(b(AP)) was not significantly different from D(b(HW)). However, analysis of variance showed a significant two-way interaction between sex and compartment model (P < 0.02), indicating that the comparisons between the sexes were different across all compartment models. The %BF calculated for the women was significantly higher than that calculated for the men by both HW and AP and for all compartment models. CONCLUSION: Our data indicate that D(b(AP)) was not significantly different from D(b(HW)). Although differences were seen in %BF between the sexes, we observed no significant differences among the compartment models within each sex for this group of older individuals.

Adipose Tissue↗

Increased fatty acid uptake and altered fatty acid metabolism in insulin-resistant muscle of obese Zucker rats.

Altered muscle fatty acid (FA) metabolism may contribute to the presence of muscle insulin resistance in the genetically obese Zucker rat. To determine whether FA uptake and disposal are altered in insulin-resistant muscle, we measured palmitate uptake, oxidation, and incorporation into di- and triglycerides in isolated rat hindquarters, as well as muscle plasma membrane fatty acid-binding protein (FABP(PM)) content of lean (n = 16, fa/+) and obese (n = 15, fa/fa) Zucker rats (12 weeks of age). Hindquarters were perfused with 7 mmol/l glucose, 1,000 micromol/l albumin-bound palmitate, and albumin-bound [1-(14)C]palmitate at rest (no insulin). Glucose uptake was 42% lower in the obese than in the lean rats and indicated the presence of muscle insulin resistance. Fractional and total rates of palmitate uptake were 42 and 74% higher in the obese than in the lean rats and were associated with higher muscle FABP(PM) content (r(2) = 0.69, P < 0.05). The percentage of palmitate oxidized was not significantly different between groups. FA disposal to storage was altered according to fiber type. When compared with lean rats, the rate of triglyceride synthesis in red muscle was 158% higher in obese rats, and the rate of palmitate incorporation into diglycerides in white muscle was 93% higher in obese rats. Pre- and postperfusion muscle triglyceride levels were higher in both red and white muscles of the obese rats. These results show that increased FA uptake and altered FA disposal to storage may contribute to the development of muscle insulin resistance in obese Zucker rats.

Animals↗

A rapid assay for detecting sulfonamides in tissues of slaughtered animals.

Governments regulate antimicrobial residues in slaughtered animals with surveillance programs for detecting drugs in food-producing animals. Although initial screening bioassay systems are recognized for their sensitivities to antimicrobial drug groups, none are sensitive to sulfonamides at or near the maximum residue levels (MRLs) in the Codex Alimentarious. We have developed a sulfonamide-sensitive rapid assay using Bacillus stearothermophilus inoculated PM indicator agar containing bromcresol purple and trimethoprim, where the end point is a combination of color change in the agar and zone of microbial growth inhibition around the sampling disk. Five sulfonamides, plus 16 other antimicrobial drugs were tested in standard concentrations in water, bovine kidney, and ground beef. Sulfonamides were detected at concentrations near the MRLs, and they were presumptively identified using para-aminobenzoic acid. The rapid assay was extremely sensitive to beta-lactams that were presumptively identified using penase. The system also was sensitive to tetracyclines, aminoglycosides, and macrolides, of which tetracyclines and gentamicin were identified using enzyme-linked immunosorbent assay (ELISA). In trials on slaughterhouse tissues submitted for testing in Ontario's meat surveillance program, the rapid assay identified twofold the number of positive kidneys and threefold the number of positive diaphragm samples compared to a standard microbiological inhibition test (MIT) currently approved. Fifty-three of 471 carcasses were sulfonamide positive with the rapid assay, while no sulfonamides were detected with the MIT. ELISA and thin-layer chromatography were used on selected samples to confirm the rapid assay sulfonamide presumptive results.

Animals↗

Frataxin activates mitochondrial energy conversion and oxidative phosphorylation.

Friedreich's ataxia (FA) is an autosomal recessive disease caused by decreased expression of the mitochondrial protein frataxin. The biological function of frataxin is unclear. The homologue of frataxin in yeast, YFH1, is required for cellular respiration and was suggested to regulate mitochondrial iron homeostasis. Patients suffering from FA exhibit decreased ATP production in skeletal muscle. We now demonstrate that overexpression of frataxin in mammalian cells causes a Ca(2+)-induced up-regulation of tricarboxylic acid cycle flux and respiration, which, in turn, leads to an increased mitochondrial membrane potential (delta psi(m)) and results in an elevated cellular ATP content. Thus, frataxin appears to be a key activator of mitochondrial energy conversion and oxidative phosphorylation.

3T3 Cells↗

Muscle palmitate uptake and binding are saturable and inhibited by antibodies to FABP(PM).

Studies show that uptake of long-chain fatty acids (LCFA) across the plasma membranes (PM) may occur partly via a carrier-mediated process and that the plasma membrane fatty acid-binding protein (FABP(PM)) may be a component of this system. To test the hypothesis that FABP(PM) is involved in transsarcolemmal transport of LCFA in muscle, we measured palmitate uptake in giant sarcolemmal vesicles and palmitate binding to PM proteins in rat muscles, (1) in the presence of increasing amounts of unbound palmitate and (2) in the absence or presence of antibody to FABP(PM). Both palmitate uptake and binding were found to be saturable functions of the unbound palmitate concentration with calculated Vmax values of 10.5 +/- 1.2 pmol/mg protein/15 sec and 45.6 +/- 2.9 nmol/mg protein/15 min and Km values of 12.8 +/- 3.8 and 18.4 +/- 1.8 nmol/L, respectively. The Vmax values for both palmitate uptake and binding were significantly decreased by 75-79% in the presence of a polyclonal antibody to the rat hepatic FABP(PM). Antibody inhibition was found to be dose-dependent and specific to LCFA. Glucose uptake was not affected by the presence of the antibody to FABP(PM). Palmitate uptake and binding were also inhibited in the presence of trypsin and phloretin. These results support the hypothesis that transsarcolemmal LCFA transport occurs in part by a carrier-mediated process and that FABP(PM) is a component of this process in muscle.

Animals↗

Cementless acetabular fixation in total hip arthroplasty using polyglycolide-lactide screws: an in vivo canine study.

Bone ingrowth into cementless acetabular components was evaluated in a canine total hip arthroplasty model, comparing components initially stabilized with polyglycolide-lactide screws with those initially stabilized with titanium screws. The acetabular shell was anchored with 2 polyglycolide-lactide screws in 16 dogs and with 2 titanium screws in 12 dogs. The dogs were followed and sacrificed at 7 weeks, 14 weeks, 10 months, or 15 months. Histomorphometric analysis of bone ingrowth into the weight-bearing dome of the acetabular shell was conducted. No difference was detected in mean bone ingrowth into the acetabular shell comparing the 2 screw groups. The results of this study do not support a significant advantage to the use of biodegradable screws for the initial stabilization of cementless acetabular components in canine total hip arthroplasty.

Acetabulum↗

Lower calorie intake enhances muscle insulin action and reduces hexosamine levels.

Previous studies have demonstrated enhanced insulin sensitivity in calorie-restricted [CR, fed 60% ad libitum (AL) one time daily] compared with AL-fed rats. To evaluate the effects of reduced food intake, independent of temporal differences in consumption, we studied AL (unlimited food access)-fed and CR (fed one time daily) rats along with groups temporally matched for feeding [fed 3 meals (M) daily]: MAL and MCR, eating 100 and 60% of AL intake, respectively. Insulin-stimulated glucose transport by isolated muscle was increased in MCR and CR vs. AL and MAL; there was no significant difference for MCR vs. CR or MAL vs. AL. Intramuscular triglyceride concentration, which is inversely related to insulin sensitivity in some conditions, did not differ among groups. Muscle concentration of UDP-N-acetylhexosamines [end products of the hexosamine biosynthetic pathway (HBP)] was lower in MCR vs. MAL despite unaltered glutamine-fructose-6-phosphate aminotransferase activity (rate-limiting enzyme for HBP). These results indicate that the CR-induced increase in insulin-stimulated glucose transport in muscle is attributable to an altered amount, not timing, of food intake and is independent of lower triglyceride concentration. They further suggest that enhanced insulin action might involve changes in HBP.

3-O-Methylglucose↗

Surgical repair for recurrent anterior instability of the shoulder.

BACKGROUND: The outcome of surgical repair for recurrent anterior instability of the shoulder at the Wellington Hospital was reviewed. METHODS: A retrospective review was undertaken of patients undergoing surgical repair for recurrent anterior instability of the shoulder at Wellington Hospital between October 1989 and November 1996. Patients were asked to complete two shoulder-specific questionnaires, and the range of motion, stability, and strength was evaluated clinically. RESULTS: A total of 37 patients (38 shoulders) who had recurrent anterior dislocation of the shoulder that was unresponsive to a physician-directed rehabilitation programme were managed with open surgical repair. Procedures included the Putti-Platt, Bristow, Magnuson-Stack, Botychev, and Bankart repairs. The mean age at the time of surgery was 24 years and the male-to-female ratio was 11.3:1. Surgery was performed on the dominant side in 63.2% of shoulders. The postoperative redislocation rate was 39.4% at an average of 4.6 years follow-up. Three patients have since required revision of their surgical repair and one patient is awaiting revision. A total of 63.2% of patients were unable to return to their previous level of sports. Differences existed between the motion in the surgically treated shoulder when compared with the contralateral side. Patients reported the most functional difficulty in throwing, working overhead, pulling, and working at shoulder level. CONCLUSIONS: The results of the present study indicate a high redislocation rate, and highlight the challenges in restoring a stable, mobile, functional shoulder.

Activities of Daily Living↗

Human cochlear expressed sequence tags provide insight into cochlear gene expression and identify candidate genes for deafness.

To identify candidate genes for human hearing disorders and to understand better human hearing at the molecular level, we constructed a human cochlear cDNA library. An aliquot of the unsubtracted cochlear library was contributed to the IMAGE Consortium at Lawrence Livermore National Laboratory for the generation of expressed sequence tags (ESTs) by the Merck/WashU EST project. Over 4000 ESTs were developed from the cochlear cDNA library and deposited in the GenBank EST database. Sequence clustering shows that the majority of clones are in low copy numbers, demonstrating the high complexity of the library. The sequences of 1388 cochlear ESTs (33%) match 517 known human genes. Among these are genes previously shown to cause both syndromic and non-syndromic hearing loss. A number of the cochlear ESTs show high homology to non-human genes, suggesting new gene family members or human homologs of animal genes. We also report the chromosomal map positions of 437 cochlear ESTs. These provide positional candidate genes for 18 different non-syndromic hearing disorders. A Human Cochlear EST Database web site (http://www.bwh.partners. org/pathology ) has been created to provide access to the cochlear clone data for gene discovery investigations.

Animals↗

Use of a polyglycolide lactide cement plug restrictor in total hip arthroplasty.

The use of a polyglycolide lactide cement plug restrictor in cemented femoral fixation during total hip arthroplasty was evaluated. Femoral cement pressurization was evaluated in vitro in a cadaveric model and the host response to polymer degradation was evaluated in vivo in a canine total hip arthroplasty model. Sixteen embalmed anatomic specimen femurs were prepared for cement femoral fixation. The intramedullary canal was plugged with either an ultrahigh molecular weight polyethylene cement plug restrictor or a polyglycolide lactide cement plug restrictor. Peak pressures in the proximal, mid, and distal portions of the cement mantle were recorded during cement insertion, cement pressurization, and implant insertion. There was no difference between the two plug groups in peak pressures throughout the cement mantle during cement insertion, pressurization, or implant insertion. Total hip arthroplasty using a cementless acetabular component and a cemented femoral stem was performed in 24 dogs. The femoral intramedullary canal was plugged with a polyethylene or a biodegradable cement plug restrictor. The dogs were sacrificed at 7 weeks, 10 months, or 15 months. Radiographically, no osteolytic lesions were seen around either plug type. Histomorphometrically, the polyglycolide lactide plugs appeared intact at 7 weeks and partially degraded by 10 and 15 months. In both plug groups, a mild fibrohistiocytic reaction with infiltration of fibrocytes, histocytes, and endothelial cells was seen. No osteolysis was observed. The results of the current study show that femoral cement pressurization can be attained in vitro using a biodegradable cement plug restrictor and that for as long as 15 months in the in vivo canine model there were no adverse reactions associated with use of these plugs compared with conventional ultrahigh molecular weight polyethylene plugs.

Animals↗

A randomized trial of hydroxyapatite coated prostheses in total hip arthroplasty.

In a prospective randomized trial, 62 consecutive primary cementless total hip arthroplasties in 55 patients were performed by one surgeon using either hydroxyapatite coated (35 hips) or nonhydroxyapatite coated femoral prostheses (27 hips). The dual tapered femoral stem had a Ti-6AI-4V plasma sprayed circumferential proximal porous coat applied to the proximal 1/3 of the stem. The middle 1/3 had a roughened blasted textured surface, and the distal 1/3 had a smooth surface. The hydroxyapatite coated femoral stems had an additional hydroxyapatite coating applied to the proximal porous coat with use of an air plasma process. The patients in the two groups were not significantly different regarding age (48.2 +/- 9.0 years hydroxyapatite group, 50.4 +/- 8.7 years control group), gender, Charnley class, or length of followup (4.4 +/- 0.7 years hydroxyapatite group, 4.9 +/- 1.0 years control group). Forty-nine patients (54 hips) were available for clinical followup, and 45 patients (50 hips) had radiographic followup. A minimum 3-year followup was recorded. To date, there have been no femoral prostheses failures. No femoral implant has migrated or subsided. Radiographically, the hydroxyapatite coated stems showed trends toward increased distal stem related cortical hypertrophy, increased cancellous condensation and less endosteal cavitation. Two nonhydroxyapatite coated stems had distal endosteal cavitation, whereas no hydroxyapatite coated stems did. There were two cases of acetabular osteolysis (revision in one) and two cases of acetabular cup migration (nonrevised), all occurring in the control group. The overall revision rate was 4%. There was no difference in Harris hip scores at 6 months (80.6 +/- 13.0 points hydroxyapatite group, 83.8 +/- 12.4 points control group) or at last followup (85.6 +/- 15.4 points hydroxyapatite group, 89.7 +/- 13.4 control group). The Harris hip pain scores also were not significantly different at 6 months or at last followup. Multiple linear regression analysis controlling for age, gender, and length of followup revealed no significant predictors of Harris hip or pain scores. The results of this study at an average of 4.6 years do not indicate a significant clinical advantage to the use of hydroxyapatite coated femoral prostheses in primary cementless total hip arthroplasty when judged by the criteria of Harris hip scores and femoral stem survivorship.

Acetabulum↗

Application of a DNA hybridization-hydrophobic-grid membrane filter method for detection and isolation of verotoxigenic escherichia coli.

Verotoxigenic Escherichia coli (VTEC) strains were isolated from food and animal fecal samples by using PCR to screen for the presence of VTEC after broth enrichment and then filtering VTEC-positive cultures through hydrophobic-grid membrane filters (HGMFs) which were incubated on MacConkey agar. The filters were probed with a digoxigenin-labeled PCR product generated by amplification of a conserved verotoxin gene sequence. Replication of the growth on filters allowed probe-positive colonies to be picked. When ground beef samples were inoculated with VTEC strains, 100% of the strains were recovered, and the detection limit was 0.1 CFU per g. Similar results were obtained with seven types of artificially contaminated vegetables. A survey of 32 packages of vegetables and 23 samples of apple cider obtained at the retail level did not reveal the presence of VTEC. However, the intestinal fecal contents of a moose, 1 of 35 wild mammals and birds examined, contained E. coli O157:H7. The DNA hybridization-HGMF method was also used in a prevalence survey of 327 raw and 744 ready-to-eat products; VTEC strains were recovered from 4.9% of the raw products and 0.7% of the ready-to-eat products. No serotype O157:H7 strains were detected. This method is particularly suited for surveys in which low numbers of VTEC-positive samples are expected and isolates are required.

Animals↗

Training-induced elevation in FABP(PM) is associated with increased palmitate use in contracting muscle.

To evaluate the effects of endurance training in rats on fatty acid metabolism, we measured the uptake and oxidation of palmitate in isolated rat hindquarters as well as the content of fatty acid-binding proteins in the plasma membranes (FABP(PM)) of red and white muscles from 16 trained (T) and 18 untrained (UT) rats. Hindquarters were perfused with 6 mM glucose, 1,800 microM palmitate, and [1-(14)C]palmitate at rest and during electrical stimulation (ES) for 25 min. FABP(PM) content was 43-226% higher in red than in white muscles and was increased by 55% in red muscles after training. A positive correlation was found to exist between succinate dehydrogenase activity and FABP(PM) content in muscle. Palmitate uptake increased by 64-73% from rest to ES in both T and UT and was 48-57% higher in T than UT both at rest (39.8 +/- 3.5 vs. 26.9 +/- 4. 4 nmol. min(-1). g(-1), T and UT, respectively) and during ES (69.0 +/- 6.1 vs. 43.9 +/- 4.4 nmol. min(-1). g(-1), T and UT, respectively). While the rats were resting, palmitate oxidation was not affected by training; palmitate oxidation during ES was higher in T than UT rats (14.8 +/- 1.3 vs. 9.3 +/- 1.9 nmol. min(-1). g(-1), T and UT, respectively). In conclusion, endurance training increases 1) plasma free fatty acid (FFA) uptake in resting and contracting perfused muscle, 2) plasma FFA oxidation in contracting perfused muscle, and 3) FABP(PM) content in red muscles. These results suggest that an increased number of these putative plasma membrane fatty acid transporters may be available in the trained muscle and may be implicated in the regulation of plasma FFA metabolism in skeletal muscle.

5'-Nucleotidase↗

Antimicrobial drug residues in milk and meat: causes, concerns, prevalence, regulations, tests, and test performance.

This paper presents a historical review of antimicrobial use in food animals, the causes of residues in meat and milk, the types of residues found, their regulation in Canada, tests used for their detection, and test performance parameters, with an emphasis on immunoassay techniques. The development of residue detection methods began shortly after the introduction of antimicrobials to food animal production in the late 1940s. From initial technical concerns expressed by the dairy industry to the present public health and international trade implications, there has been an ongoing need for reliable, sensitive, and economical methods for the detection of antimicrobial residues in food animal products such as milk and meat. Initially there were microbial growth inhibition tests, followed by more sensitive and specific methods based on receptor binding, immunochemical, and chromatographic principle. An understanding of basic test performance parameters and their implications is essential when choosing an analytical strategy for residue testing. While each test format has its own attributes, none test will meet all the required analytical needs. Therefore the use of a tiered or integrated system employing assays designated for screening and confirmation is necessary to ensure that foods containing violative residues are not introduced into the food chain.

Animals↗

Characteristics of the LacTek test as applied to tissue samples: assessment of performance using incurred field samples.

The Lactek test, marketed for antimicrobial residue detection in milk, was validated for the detection of antimicrobial residues in tissues. A previous study found that the LacTek test could confidently identify tissue samples spiked with antimicrobial residues. However, the test could not reliably distinguish violative from nonviolative spiked samples relative to Canadian maximum residue limits (MRLs). The objectives of this study were to assess and compare the performance of the LacTek tests for beta-lactams, tetracyclines, gentamicin, and sulfamethazine on samples containing naturally incurred residues by running the test in parallel with the standard microbial inhibition test (MIT) presently used for the routine testing of tissues at our facility and to assess the agreement with high pressure liquid chromatographic (HPLC) determinative methods. Parallel testing with the official MIT found that the Lactek tests could be confidently used for testing tissue samples containing incurred residues. Among 1,008 MIT-positive samples, the LacTek test found that 90% contained beta-lactams and/or tetracyclines. A further 7.3% of violative residues could not be identified to an antimicrobial class. In addition, 9% of samples testing negative on the MIT were found to contain an antimicrobial residue by the LacTek tests. Comparative testing with HPLC methods found that there was very good agreement between the two tests and that most violations were due to penicillin G and oxytetracycline. Although the LacTek test cannot be used to distinguish violative from nonviolative residue levels, it does offer several advantages over the present MIT. These include speed, ease of use, the ability to identify residues to a specific class, and an improved sensitivity at the MRL level for the most commonly found antimicrobials in tissue.

Anti-Bacterial Agents↗

Mitomycin-induced synthesis of a Shiga-like toxin from enteropathogenic Escherichia coli H.I.8.

Escherichia coli H.I.8, an O128 infant diarrhea isolate, produces low titers of a unique Shiga-like toxin (SLT), called SLT-IIva, which is a variant of SLT-II. We investigated induction of toxin synthesis and the putative association of a bacteriophage with toxin synthesis. Induction of broth cultures of strain H.I.8 with mitomycin yielded a 3,000-fold increase in SLT-IIva, production of a colicin, and appearance of a bacteriophage. Southern hybridization demonstrated that the genes for SLT-IIva were not carried by the bacteriophage.

Bacterial Toxins↗

Validation of the LacTek test applied to spiked extracts of tissue samples: determination of performance characteristics.

LacTek tests are competitive enzyme-linked immunosorbent assays intended for rapid detection of antimicrobial residues in bovine milk. In this study, the LacTek test protocol was modified for use with extracts of bovine tissue to detect beta-lactam, tetracycline, and sulfamethazine residues. Test performance characteristics--precision, accuracy, ruggedness, practicability, and analytical specificity and sensitivity--were investigated. Results suggest that LacTek tests can be easily adapted to detect antimicrobial residues in extracts of lean ground beef. However, positive samples may not contain residues at violative concentrations (i.e., Canadian maximum residue limits), and therefore, additional analysis would be required for final confirmation and quantitation (e.g., chromatography).

Animals↗