Comparison of computer-based and paper-based imagery strategies in learning anatomy.

This study evaluated the use of computer-based interactive imagery on students' achievement scores when compared with paper-based static imagery. It also assessed students' perceptions about the two imagery strategies and their different components. Sixty-four freshmen veterinary students (50 females, 14 males), enrolled in a comparative anatomy course, volunteered to participate in the study. This study used a pretest/posttest comparison group design and data was examined by analysis of covariance (ANCOVA). A close-ended questionnaire was administered to collect students' perceptions about the two imagery strategies. The mean difference in students' perceptions between the two strategies was analyzed using a two-tailed paired t-test. No significant differences were observed between computer-based interactive imagery and paper-based static imagery in the immediate recall of anatomical information. There was a significant difference in students' opinions toward the two strategies: students perceived computer-based interactive imagery as a better strategy in the assimilation of anatomical information than paper-based static imagery.

Using computer-based interactive imagery strategies for designing instructional anatomy programs.

In an effort to design and implement effective anatomy educational programs, this study was conducted to evaluate students' perceptions toward using two computer-based self-directed instructional modules (e.g., digestive system and canine skull) that were designed utilizing interactive imagery strategy for teaching and learning veterinary anatomy. Sixty-eight freshmen veterinary students and one graduate student participated in this study. Open-ended and close-ended questionnaires were administered to evaluate the utilization of computer-based interactive imagery strategy in developing anatomy instructional programs, and to collect data about the students' perceptions toward the use of interactive images in teaching and learning of anatomy. Means and standard deviations were calculated and analyzed for close-ended items. The open-ended questionnaire items were analyzed to identify shared patterns or themes in the students' experience after using the two instructional anatomy modules. Students reported positive attitudes toward the interactive imagery strategy used in the development of computer-based anatomy modules. Based on our findings, this study outlines the characteristics of effective instructional images that will serve as guidelines for the preparation and selection of anatomical images, as well as, how to utilize these images to develop computer-based instructional anatomy programs. Students perceived interactive imagery as an effective design strategy that helped them learn anatomical concepts.

Studies on the mechanism of diarrhoea induced by Escherichia coli heat-stable enterotoxin (STa) in newborn calves.

Enterotoxigenic Escherichia coli (ETEC) produces a heat-stable enterotoxin (STa) that binds to and activates a putative intestinal receptor, guanylate cyclase, causing an increase in the intracellular levels of cyclic guanosine monophosphate (cGMP). Using flow cytometry and 125I-STa binding assays, we studied the distribution of STa-receptors on enterocytes isolated from different segments of the newborn calf's intestinal tract. We also investigated the effect of STa on the intracellular levels of cGMP and ion transport to the intestinal lumen. More STa-receptors were found on enterocytes prepared from the ileum than on enterocytes obtained from the other segments of the intestinal tract. Guanylate cyclase activity was higher in the ileum of STa-challenged calves than in the ileum of control calves. No changes were observed in the guanylate cyclase activity of the other intestinal segments of the STa-challenged and control calves. Na+ levels, as measured by atomic absorption spectroscopy, were significantly increased in the luminal contents of the ileum of STa-challenged calves, whereas serum Cl- levels were significantly lower in the STa-challenged calves than in control calves. This study supports previous observations on the role of guanylate cyclase in the initiation of STa-induced secretory diarrhoea and suggests that Na+/Cl- coupling may be the major mechanism for the loss of ions in the diarrhoeal response that is mostly induced in the ileum of newborn calves.

Characterization of the interaction of Escherichia coli heat-stable enterotoxin (STa) with its putative receptor on the intestinal tract of newborn calves.

Enterotoxigenic Escherichia coli (ETEC) induces severe diarrhea in newborn calves through the elaboration of heat-stable enterotoxin (STa). We investigated the distribution and characteristics of the STa-specific receptors on enterocytes and brush border membrane vesicles (BBMVs) prepared from anterior jejunum, posterior jejunum, ileum and colon of newborn calves. We found that density of the receptors and their affinity to STa were higher on enterocytes and BBMVs that were derived from the ileum than enterocytes and BBMVs prepared from other segments of the calf intestine. This study suggests that, in newborn calves, the ileum is the major part of the intestinal tract that is affected in the course of secretory diarrhea caused by STa-producing ETEC strains.

Application and evaluation of the alamarBlue assay for cell growth and survival of fibroblasts.

Cell proliferation assays are essential to developing an understanding of the molecular mechanisms that modulate cell growth and differentiation. In this paper, we describe the application of alamarBlue, a new and versatile metabolic dye, for the detection of Swiss 3T3 fibroblast proliferation and/or survival. As a redox indicator, alamarBlue is reduced by reactions innate to cellular metabolism and, therefore, provides an indirect measure of viable cell number. Various assay parameters were optimized for a 96-well format to achieve a detectable range of fibroblast cell number from 100 to 20,000 cells/well, which is similar to that obtained with traditional (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) and [3H]thymidine assay techniques. Standard (reference) curves generated with a known fibroblast stimulator were used to facilitate quantitation and comparison of unknown test substances. The alamarBlue assay offers the advantages of technical simplicity, freedom from radioisotopes, versatility in detection, no extraction, and excellent reproducibility and sensitivity. We anticipate that this simple and versatile alamarBlue assay, when used alone or in conjunction with other bioassays, will be a useful tool for investigating the complex mechanisms of cellular proliferation.

Effect of betamethasone and exercise on equine carpal joints with osteochondral fragments.

Osteochondral fragments were created arthroscopically on the distal aspect of both radial carpal bones in 12 horses. On day 14 after surgery, one middle carpal joint of each horse was injected with 2.5 mL Betavet Soluspan (3.9 mg betamethasone sodium phosphate and 12 mg betamethasone acetate per milliliter) and the contralateral joint was injected with 2.5 mL saline as a control. Intra-articular treatments were repeated on day 35. On day 17, six horses began exercising 5 days per week on a high-speed treadmill. The other six horses were kept in box stalls throughout the study as nonexercised controls. On day 56, all horses were examined clinically and radiographically and then were euthanatized. Samples were obtained for histological, histochemical, and biochemical evaluation. Mild lameness was observed in five of the six exercised horses at day 56; four horses were lame in the control limb and one horse was lame in the treated limb. Of the five nonexercised horses evaluated for lameness, two were lame in the control limb, two were lame in the treated limb, and one was lame in both the control and the treated limb. No differences were noted on radiographs or palpation of steroid treated limbs versus control limbs. Firm reattachment of the osteochondral fragment to the radial carpal bone occurred in all but three joints. Gross cartilage damage was not different between steroid-treated joints and joints injected with saline. Histologically, there were no significant detrimental effects of beta-methasone with or without exercise, but there was a tendency for more pathological change in treated joints. No significant difference in the water content or uronic acid concentration was detected between treated and control joints. Intra-articular betamethasone administration in this carpal chip model was not associated with any significant detrimental effects in either rested or exercised horses.

Effects of difloxacin on the metabolism of glycosaminoglycans and collagen in organ cultures of articular cartilage.

Fluoroquinolones, including difloxacin, are potent antibacterial compounds which, as a side effect, cause lesions in articular-epiphyseal cartilage complexes (AECC) of growing animals. To evaluate the effects of difloxacin on the structure of AECC and the metabolism of sulfated glycosaminoglycans (GAG) and collagen, explants of AECC were obtained from 18 healthy, 3-month-old Beagle dogs and cultured in medium which either had no difloxacin or had the drug at one of three concentrations (40, 80, or 160 micrograms/ml). Rates of synthesis of GAG and collagen were reduced by concentrations of difloxacin that were at or above 80 micrograms/ml. The rate of synthesis of total protein, however, was reduced only at the highest dose level. Catabolism of GAG and collagen was unaffected by the treatment. The principal ultrastructural changes in affected chondrocytes were distension of rough endoplasmic reticulum with electron-dense material that was probably protein, and vacuolation of cytoplasm. Structural changes were not observed in the extracellular matrix. It, therefore, appeared plausible that difloxacin affected chondrocytes by interfering with secretion of the matrix components, GAG and collagen.

Immunolocalization of epidermal growth factor (EGF) and EGF receptors in the porcine upper gastrointestinal tract.

The factors controlling growth and maturation in the porcine gastrointestinal tract are not well understood. Epidermal growth factor (EGF) is a polypeptide that has been implicated in the control of gastrointestinal tract growth, maturation, and protection in other species. Immunoreactive EGF (IR-EGF) and EGF receptors (EGF-R) were histochemically identified in formalin-fixed tissues of the upper digestive tract of 1-, 3-, 7-, 14-, 21-, and 28-day-old pigs. The ductal epithelium consistently contained IR-EGF in the parotid salivary gland of pigs of all ages and in the mandibular salivary gland in pigs greater than or equal to 7 days old. Immunoreactive EGF was detected in the mucosal epithelium of the esophagus and nonglandular portion of the stomach, and in the pancreas and liver in all pigs. Gastric gland IR-EGF was inconsistently detected in pigs less than 14 days old and was consistently observed in all older pigs. Enterocyte EGF immunoreactivity was usually weak and was variably detected in the duodenum of pigs less than or equal to 7 days old and in the jejunum of pigs less than or equal to 14 days old, but was consistently observed in older pigs. Ileal immunoreactivity was erratic. Immunoreactive EGF-R were identified in the esophageal epithelium of all pigs, and in the nonglandular gastric and glandular gastric mucosa of all pigs, except for two 7-day-old pigs and one 7-day-old pig, respectively. Immunoreactive EGF-R were detected in the duodenal, jejunal, and ileal enterocytes of pigs of all ages examined.

Effects of intra-articular administration of methylprednisolone acetate on normal equine articular cartilage.

The effects of the corticosteroid 6-alpha-methylprednisolone acetate on normal equine articular cartilage were evaluated, using the middle carpal joint in 4 clinically normal young horses. One middle carpal joint of each horse was injected 3 times with 100 mg of 6-alpha-methylprednisolone acetate, at 14-day intervals. The opposite middle carpal joint (control) was injected with 2.5 ml of lactated Ringer solution at the same intervals. Effects were studied until 8 weeks after the first injection. Evaluation included clinical and radiographic examination, and gross, microscopic, and biochemical evaluation of joint tissues. Horses remained clinically normal during the study, and significant radiographic changes were not observed. Safranin-0 matrix staining intensity and uronic acid content were significantly (P less than 0.05) lower and hydroxyproline content was significantly (P less than 0.05) higher in articular cartilage of corticosteroid-injected joints vs control joints.

Effect of orally administered epidermal growth factor on the jejunal mucosa of weaned pigs.

The effect of ingested epidermal growth factor (EGF) on the small intestinal mucosa of conventionally weaned pigs was determined. At 21 days of age, 39 pigs were randomly distributed into suckling and weaned treatment groups that were administered 124 micrograms of EGF, 372 micrograms of EGF, or the dosing compound daily. Fecal water content was determined daily. On postweaning days 0, 3, 6, and 9, representative pigs from each group were euthanatized, and jejunal mucosa samples were collected for determination of villus-to-crypt ratio, total protein content, disaccharidase activities, and microbiological populations. At postweaning day 3, the 372-micrograms dose of EGF significantly (P less than or equal to 0.05) increased jejunal lactase and sucrase activities in the weaned pigs. Increased lactase activity was not greater than that of the suckling pig controls, whereas sucrase activity was significantly (P less than or equal to 0.05) higher than that of the suckling pig controls. Significant changes were not observed in villus-to-crypt ratio, mucosal protein content, or disaccharidase activities on other collection days.

Lectin binding to small intestinal goblet cells of newborn, suckling, and weaned pigs.

Lectin binding of small intestinal goblet cells was examined in newborn, suckling, and weaned pigs. Sections of duodenum, proximal portion of the jejunum, distal portion of the jejunum, and ileum were embedded in a hydrophilic acrylic resin and treated with each of the following lectins: Canavalia ensiformis, Ricinus communis I, Glycine max, Ulex europaeus I, and Triticum vulgaris. Percentages of goblet cells binding each lectin were calculated within intestinal regions. Differences in lectin-binding affinity were detected among pigs of various ages and among various intestinal regions within pig age groups.

Effect of butorphanol, pentazocine, meperidine, or metoclopramide on intestinal motility in female ponies.

Effect of butorphanol, pentazocine, meperidine, and metoclopramide on jejunal and pelvic flexure myoelectric and mechanical activity in 4 female ponies was investigated. The agent to be tested or saline solution was administered IV at the start of a 6-hour recording trial. In the jejunum, duration between activity fronts of regular spiking activity, defined as the length of the migrating myoelectric complex (MMC), was measured. The average duration of the MMC during control trials was 150 +/- 46 minutes. The average duration of the MMC after meperidine, butorphanol, pentazocine, and metoclopramide administration was 295 +/- 70 minutes, 260 +/- 60 minutes, 275 +/- 60 minutes, and 163 +/- 64 minutes, respectively. Meperidine, butorphanol, or pentazocine significantly increased the MMC duration (P less than 0.05), and did not significantly alter the pelvic flexure activity. Seemingly, meperidine, butorphanol, and pentazocine inhibited cyclic myoelectric activity in the jejunum. Metoclopramide had no effect on jejunal or pelvic flexure motility.

Growth-stimulating substances in porcine milk.

Colostral and milk samples were collected from 6 primiparous sows on postpartum days 0, 9, 18, and 27. Insulin content was measured and epidermal growth factor (EGF) concentration was determined by use of radioimmunoassay and radioreceptor assay, respectively. Total milk protein concentration was measured colorimetrically. Concentrations of insulin, EGF, and protein in the colostral samples were significantly (P less than 0.01) higher than those found in the milk. Milk samples obtained from postpartum days 9, 18, and 27 did not differ significantly in insulin, EGF, or total protein content.

Ultrastructure of equine endothelial cells exposed to endotoxin and flunixin meglumine and equine neutrophils.

An in vitro system of cultured equine endothelial cells was evaluated as a model for endotoxin (ET) exposure in the horse. Primary cell lines from pulmonary vessels and aortas were cultured from tissues of 6 horses. Effects of ET alone with and without serum and in combination with the cyclo-oxygenase inhibitor flunixin meglumine and isolated equine neutrophils were evaluated by transmission electron microscopy. Cells plus serum were incubated with 10, 25, 50, or 100 micrograms of ET/ml of incubation medium for 1, 3, 8, or 24 hours. Cells without serum were cultured for 1 and 3 hours. Flunixin meglumine was used at a concentration of 20 micrograms/ml. Cells also were incubated in the presence of 1,000, 5,000, or 20,000 neutrophils/ml plus ET and in the presence of a combination of ET and flunixin meglumine for 1 or 3 hours. Endotoxin alone did not cause cell damage, and the only evidence of an effect was an increased number of secondary lysosomes at incubation hour 8. At incubation hour 24, cells appeared normal. Endotoxin plus neutrophils caused cells to become round and detach from the growth substrate. Cell pathologic changes included swollen and distorted mitochondria and cytoplasmic vacuolization. Response to the ET plus neutrophil combination was variable and ranged from 5% to 50% of the cells being affected. The variability appeared to have some correlation with cell age, as well as individual preparation of neutrophils.

In vitro characterization of porcine juvenile articular cartilage.

Dispersed cell cultures were established from the articular cartilage of the proximal portion of the humerus of young pigs. Articular and epiphyseal portions of the cartilage were separated, minced, and enzymatically dispersed, using bacterial collagenase. Morphologically, 2 cell types were observed, using phase-contrast microscopy. Smaller polygonal cells (32.5 +/- 3.5 microns diameter) containing cytoplasmic granules were found in both areas of the cartilage. In cultures from the articular region, cells grew as monolayer cultures and initially did not demonstrate contact inhibition. In cultures from the epiphyseal region, cells grew in a multilayered manner in a colonial arrangement with cells being released from the center of the colony into the culture medium. Small granular particles (0.03 to 0.08 micron diameter) were secreted by cells in both culture systems. Particle secretion was greater in epiphyseal cultures than in articular cultures with the rate decreasing as confluency was approached. These particles stained positively for lipid and alkaline phosphatase. Acridine orange was also incorporated into the granules. The 2nd cell type, a stellate-shaped cell (60 +/- 7.6 micron diameter), was found mainly surrounding the outside of colonial areas in epiphyseal cultures. These cells did not secrete small granular particles and stained positive for factor VIII. Evaluation of cultures by scanning and transmission electron microscopy further supported the presence of 2 cell types. With scanning electron microscopy, the smaller polygonal cell was characterized by varying sizes of blebs (0.03 to 0.1 micron diameter) associated with the cell membrane and small cytoplasmic processes projecting from the cell's surface.(ABSTRACT TRUNCATED AT 250 WORDS)

Equine endothelial cells in vitro.

Certain in vitro culture conditions were determined for equine endothelial cells obtained from the aorta and pulmonary arteries. Cells were enzymatically isolated from the vessel lumen, using clostridial collagenase (2.5 mg/ml of Hanks's balanced salt solution) incubated at 37 C for 30 minutes. Cells were cultured in alpha minimum essential medium supplemented with plasma-derived and nonplasma-derived bovine fetal sera, endothelial cell-growth supplement, heparin, and antibiotics. Smooth muscle cell growth was not inhibited with nonplasma-derived animal sera, plasma-derived equine serum, or heparin. Heparin and a serum replacement were toxic to the cells used in the present study. Statistically significant differences were not found between the various media supplements.

Electromyographic, myomechanical, and intraluminal pressure changes associated with acute extraluminal obstruction of the jejunum in conscious ponies.

Bipolar electrodes, strain gauge force transducers, intraluminal pressure recording catheters, and extraluminal intestinal obstructors were surgically implanted in 4 ponies to record myoelectrical and mechanical activity of the distal portion of the jejunum and ileum. After determining normal intestinal activity and pressures, the distal portion of the jejunum was obstructed with an extraluminal obstructor. Myoelectrical and mechanical activity recorded from jejunal segments proximal to the obstruction increased significantly (P less than 0.01), whereas activity distal to the obstruction remained unchanged. Intraluminal pressure increases were recorded during periods of intestinal spasm. Obstruction pressures remained unchanged from preobstruction pressures.

Endotoxin-induced eicosanoid production by equine vascular endothelial cells and neutrophils.

Dispersed equine vascular endothelial cells grown in tissue culture, and freshly isolated neutrophils were used to determine direct effects of endotoxin on cyclooxygenase and lipoxygenase products. Endothelial cells (10(7)/ml) or neutrophils (2 X 10(6)/ml) were incubated with (a) buffer, (b) endotoxin (10 micrograms/ml), (c) endotoxin + flunixin meglumine (10 micrograms/ml), or (d) calcium ionophore, A23187 (10 micrograms/ml). Thromboxane (TxB2), prostacyclin (6-keto-PGF1 alpha), and leukotriene C4 (LTC4) were determined in the incubation fluid by radioimmunoassay. Thromboxane and prostacyclin levels increased in endothelial cells incubated with endotoxin. Treatment with flunixin meglumine prevented the endotoxin-induced release of these cyclooxygenase products to levels below those observed in control cells. Leukotriene production was increased in endothelial cells incubated with endotoxin plus flunixin meglumine. Endotoxin as well as endotoxin plus flunixin meglumine increased the production of prostacyclin and LTC4 by freshly isolated neutrophils. Cells exposed to endotoxin plus flunixin meglumine produced more LTC4 than cells exposed to endotoxin. The data revealed that endotoxin has a direct effect on arachidonic acid metabolism in endothelial cells and neutrophils. Flunixin meglumine reduced the level of cyclooxygenase products but increased the level of lipoxygenase products. Therefore, the well-established beneficial effects of cyclooxygenase inhibitors during endotoxemia may be improved even more if they are used in conjunction with lipoxygenase inhibitors or a combined cyclooxygenase-lipoxygenase inhibitor.

Motility of the distal portion of the jejunum and pelvic flexure in ponies: effects of six drugs.

Bipolar stainless steel electrodes were surgically implanted in 4 ponies to record myoelectrical and mechanical activity of the distal portion of the jejunum and pelvic flexure. After determining normal activity, the effects of neostigmine, xylazine, flunixin meglumine, dipyrone, panthenol, and atropine sulfate were determined. Flunixin meglumine, dipyrone, and panthenol had no effect on the motility of the jejunum or pelvic flexure. Xylazine and atropine sulfate decreased motility of the distal portion of the jejunum and pelvic flexure, with atropine sulfate having a greater effect and lasting longer. Neostigmine stimulated propulsive motility in the pelvic flexure only.