Reciprocal regulation between TOC1 and LHY/CCA1 within the Arabidopsis circadian clock.

The interactive regulation between clock genes is central for oscillator function. Here, we show interactions between the Arabidopsis clock genes LATE ELONGATED HYPOCOTYL (LHY), CIRCADIAN CLOCK ASSOCIATED 1 (CCA1), and TIMING OF CAB EXPRESSION 1 (TOC1). The MYB transcription factors LHY and CCA1 negatively regulate TOC1 expression. We show that both proteins bind to a region in the TOC1 promoter that is critical for its clock regulation. Conversely, TOC1 appears to participate in the positive regulation of LHY and CCA1 expression. Our results indicate that these interactions form a loop critical for clock function in Arabidopsis.

Photoreceptor degeneration preceding clinical scrapie encephalopathy in hamsters.

Hamsters experimentally inoculated with scrapie had histologic and ultrastructural changes in their retinas before clinical signs of central nervous system disease became evident at 50 days postinoculation. The retinal changes were limited to the photoreceptors and began with degenerative changes in the inner segments as early as 8 days postinoculation. As the inner segment lesions became more prominent, associated degenerative changes within outer segments were seen (38 days postinoculation). An influx of macrophages was associated with increasing degenerative changes; these cells engulfed cellular debris did not appear to contribute directly to the degenerative process. Retinas having the greatest damage to the inner segment area also had moderate reduction in outer nuclear layer density and outer segment length, and in numbers of outer segments. All of the retinal changes described occurred almost exclusively in the central to mid-region of retina with relative sparing of peripheral retina.

Retinal degeneration during clinical scrapie encephalopathy in hamsters.

Weanling hamsters were inoculated intracerebrally with brain suspensions from normal or scrapie-infected hamsters. A third group of uninoculated animal was fed cuprizone. Histologic and electron microscopic examination of the neural retinas and retinal pigment epithelium was done. At 50 days postinoculation, when scrapie-infected animals began to show clinical signs of encephalopathy, there was a variable degree of photoreceptor degeneration. By the time animals were moribund, at 74 days postinoculation, photoreceptor degeneration was severe, as demonstrated by loss of most outer and inner segments and cell bodies. The outer plexiform, inner nuclear, and inner plexiform layers were reduced in thickness. Some retinal pigment epithelial cells contained lipoidal inclusions. The neural retinas and retinal pigment epithelium of noninfected and cuprizone-treated animals were normal. We have previously shown that the scrapie agent accumulates in the retina; that together with our present work, we conclude that the scrapie agent is the cause of photoreceptor degeneration in experimentally inoculated hamsters.

A Monte Carlo dosimetry-based evaluation of the 7Li(p,n)7Be reaction near threshold for accelerator boron neutron capture therapy.

Advanced methods of boron neutron capture therapy (BNCT) use an epithermal neutron beam in conjunction with tumor-targeting boron compounds for irradiation of glioblastomas and metastatic melanomas. A common neutron-producing reaction considered for accelerator-based BNCT is 7Li(p,n)7Be, whose cross section increases very rapidly within several tens of keV of the reaction threshold at 1.88 MeV. Operation in the proton energy region near threshold will have an appreciable thick target neutron yield, but the neutrons produced will have relatively low energies that require little moderation to reach the epithermal range desirable for BNCT. Because of its relatively low projected accelerator cost and the portability of the neutron source/target assembly, BNCT based on the near-threshold technique is considered an attractive candidate for widespread hospital use. A systematic Monte Carlo N-Particle (MCNP) investigation of the dosimetric properties of near-threshold neutron beams has been performed. Results of these studies indicate that accelerator proton energies between 1.93 and 1.99 MeV, using 5 cm of H2O moderator followed by thin 6Li and Pb shields, can provide therapeutically useful beams with treatment times less than one hour and accelerator currents less than 5 mA.

Donut-shaped "miniparticles" in nuclei of human and rat cells.

Donut-shaped "miniparticles" were extracted from nuclei of various types of human and rat cells. Electron-microscopic investigation showed these particles were predominantly in sucrose density gradient fractions that had an approximate sedimentation coefficient of 21S. These particles were 113 +/- 8A in diameter and had an electron dense center of 29 +/- 6A. They appeared to be composed of 8 subunits. Quantitative analysis of the number of these particles by electron-micrographic field counting showed nuclei of tumor samples had a larger amount of the particles than the cytosol. However, normal cell cytosol had a larger number of particles than the nuclei. A group of proteins in the 25,000-33,000 molecular weight range was shown to be the main protein component by two dimensional gel electrophoresis.

Retrospective evaluation of equine prepurchase examinations performed 1991-2000.

REASONS FOR PERFORMING STUDY: Prepurchase examinations of horses are used increasingly as a means of evaluating future soundness. Data is lacking in the literature of the radiographic findings and results of the lameness examination of comprehensive prepurchase examinations. OBJECTIVE: To summarise the clinical and radiographic findings of prepurchase examinations and determine if radiographic findings correlated with the lameness examination and sale outcome. METHODS: Records of 510 cases were reviewed, radiographs evaluated and grades assigned the navicular bone, distal phalanx, and tarsus. Follow-up information on the horse status was obtained by telephone interviews for 173 horses. RESULTS: Thoroughbred geldings represented the most common breed and sex, mean age 8 years, mean asking price 12,439.40 dollars, and 52.8% were lame. Radiography was the most common diagnostic procedure performed (61.6%), with views of the front feet requested most often (86.6%) followed by the tarsi (68.1%). Grade 1 was most common for the navicular bone while Grade 2 predominated for the distal phalanx. The number of sound horses decreased as grades became more severe. For the tarsi, Grades 0 and 1 were most common for the proximal intertarsal and distal intertarsal/metatarsal joints, respectively. Horses with significant tarsal changes were still able to compete at their expected level. With respect to the radiographic examination, the mean +/- s.d. grade of the horses which were not lame at follow-up was 1.2 +/- 0.9 for the navicular bone and 15 +/- 0.8 for the third phalanx. The mean +/- s.d. grade of sound horses for the distal intertarsal joint was 0.7 +/- 0.6 and 1.14 +/- 0.8 for the tarsometatarsal joint. Horses for which owner follow-up was available and which had a Grade 3 score were also evaluated. For the navicular bone, 17/31 with a Grade 3 remained in active use at follow-up and for the distal phalanx 21/27 were in active use. For the distal intertarsal and tarsometatarsal joints, 20/21 with a Grade 3 were still in active use. CONCLUSIONS: Prepurchase examinations can have a significant effect on the outcome of the sale. For the navicular bone and distal phalanx, higher grades were associated with lameness. In contrast, higher grades in the tarsus were less likely to be associated with lameness. Warmbloods tended to have more extensive changes in the navicular bone and distal phalanx relative to Thoroughbreds but were not as lame. POTENTIAL RELEVANCE: Radiographic changes detected in the navicular bone, distal phalanx and tarsus should be interpreted with consideration to the clinical examination.

In vitro investigation of the interaction between nitric oxide and cyclo-oxygenase activity in equine ventral colon smooth muscle.

The objective of this study was to determine if a correlation exists between the presence of nitric oxide and prostaglandin release in the equine ventral colon smooth muscle, since this relationship may accentuate the inflammatory process during intestinal injury. Tissue was collected from the ventral colon, cut into muscle strips oriented along the circular, longitudinal and taenial layers, and mounted in a tissue bath system. Samples of the bath fluid were collected before, following electrical field stimulation (EFS), and following EFS in the presence of L-NAME, a nitric oxide synthase inhibitor. Muscle strips were also obtained following systemic administration of a cyclo-oxygnease inhibitor and samples were collected using the previously described protocol. Concentrations of prostaglandins were determined in the fluid samples using an ELISA. Electrical field stimulated release of nitric oxide produced a significant increase in prostaglandin production which did not occur in the presence of L-NAME. Systemic administration of flunixin meglumine reduced prostaglandin levels at all sampling periods, although a small increase was present following EFS. The results of this study support the hypothesis that there is a correlation between the release of nitric oxide and the production of prostaglandins in the smooth muscle of the large colon. This association between nitric oxide and prostaglandin may act as an important regulatory mechanism for various physiological mechanisms, such as vascular smooth muscle tone, and may contribute to amplified tissue injury when the induced forms of both enzymes are activated during an inflammatory insult. This suggests that the use and development of COX2 and iNOS inhibitors may help attenuate the inflammatory response following intestinal injury.

Microvascular circulation of the descending colon in horses.

The microvascular circulation of the descending colon was studied in 5 adult horses, using microangiography and light microscopy combined with gross studies and scanning electron microscopy of vascular replicas. After heparinization, horses were euthanatized, and 3 segments of the descending colon and its mesentery containing 1 vascular arcade were removed from each horse. The fecal balls were gently massaged from the lumen, and the blood was flushed free of the circulation with isotonic NaCl. In 5 segments, the vascular system was injected with a modified radiopaque medium and evaluated radiographically. Specimens examined radiographically also were prepared for histologic examination, using standard methods. Ten segments were injected with 1 of 2 types of plastics and studied grossly or by scanning electron microscopy. Arcuate arteries gave rise to a descending colonic rete that surrounded the vein and supplied numerous descending colonic lymph nodes. The rete also supplied the mesocolon and the descending colonic tissue. Short filamentous vessels arising from the rete directly penetrated the mesenteric tenia to supply an intermuscular plexus between the longitudinal and circular muscle layers of the muscularis externa. Larger vessels arising from either side of the rete divided into the long- and short-terminal arteries that supplied an extensive submucosal plexus, which was continuous around the circumference. The submucosal plexus supplied the mucosa, the tunica muscularis, and the serosa. Vessels running centrifugally from the submucosal plexus formed an intermuscular plexus between the longitudinal and circular muscle layers of the muscularis externa. The intermuscular plexus at the mesenteric angle also was supplied by vessels branching from the short-terminal arteries as they penetrated the muscularis externa.(ABSTRACT TRUNCATED AT 250 WORDS)

Jejunal microvasculature of the llama and alpaca.

The vasculature of the jejunum was studied in 6 llamas and 1 alpaca, using a combination of microangiography, standard light microscopy, and vascular cast imaging. The casts were examined by use of scanning electron microscopy and low-power dissecting microscopy. After administration of 40,000 IU of heparin, all animals were euthanatized by administration of an overdose of sodium pentobarbital. Three sections of jejunum and their respective arcuate vessels were isolated from each animal. One section was immediately placed in formalin for later H&E staining. The second and third sections were placed in warm saline solution, and the vasculature was flushed free of all blood by repeated infusions of the solution. Once flushed of all blood, one section was infused with a radio-opaque medium and subsequently evaluated by microangiography, and the remaining section was perfused with a methylmethacrylate polymer for creation of vascular casts. The arcuate vessels branched into extensive primary and secondary arcades prior to giving rise to the marginal rete. Muscular arteries and small veins left the marginal rete and penetrated the tunica serosa and tunica muscularis to provide nutrients or drain the mesenteric angle, respectively, or entered into the circumferential submucosal network. The primary penetrating vessels in the submucosa formed an extensive submucosal plexus that supplied the tunica serosa, tunica muscularis, and tunica mucosa. The primary penetrating vessels anastomosed with vessels from oral and aboral sections and with their counterparts from the opposite side at the antimesenteric border.(ABSTRACT TRUNCATED AT 250 WORDS)

In vitro investigation of the effect of prostaglandins and nonsteroidal anti-inflammatory drugs on contractile activity of the equine smooth muscle of the dorsal colon, ventral colon, and pelvic flexure.

OBJECTIVES: To determine the in vitro effect of prostaglandin E2 (PGE2), PGF2alpha, PGI2; and nonsteroidal anti-inflammatory drugs (NSAID; ie, flunixin meglumine, ketoprofen, carprofen, and phenylbutazone) on contractile activity of the equine dorsal colon, ventral colon, and pelvic flexure circular and longitudinal smooth muscle. ANIMALS: 26 healthy horses. PROCEDURE: Tissue collected from the ventral colon, dorsal colon, and pelvic flexure was cut into strips and mounted in a tissue bath system where contractile strength was determined. Incremental doses of PGE2, PGF2alpha,, PGI2, flunixin meglumine, carprofen, ketoprofen, and phenylbutazone were added to the baths, and the contractile activity was recorded for each location and orientation of smooth muscle. RESULTS: In substance P-stimulated tissues, PGE2 and PGF2alpha enhanced contractility in the longitudinal smooth muscle with a decrease or no effect on circular smooth muscle activity. Prostaglandin I2 inhibited the circular smooth muscle response with no effect on the longitudinal muscle. The activity of NSAID was predominantly inhibitory regardless of location or muscle orientation. CONCLUSIONS AND CLINICAL RELEVANCE: In the equine large intestine, exogenous prostaglandins had a variable effect on contractile activity, depending on the location in the colon and orientation of the smooth muscle. The administration of NSAID inhibited contractility, with flunixin meglumine generally inducing the most profound inhibition relative to the other NSAID evaluated in substance P-stimulated smooth muscle of the large intestine. The results of this study indicate that prolonged use of NSAID may potentially predispose horses to develop gastrointestinal tract stasis and subsequent impaction.

Role of nitric oxide in in vitro contractile activity of the third compartment of the stomach in llamas.

OBJECTIVE: To determine the role of nitric oxide and an apamin-sensitive nonadrenergic-noncholinergic inhibitory transmitter in in vitro contractile activity of the third compartment in llamas. SAMPLE POPULATION: Isolated strips of third compartment of the stomach from 5 llamas. PROCEDURE: Strips were mounted in tissue baths containing oxygenated Kreb's buffer solution and connected to a polygraph chart recorder to measure contractile activity. Atropine, guanethidine, and indomethacin were added to tissue baths to inhibit muscarinic receptors, adrenoreceptors, and prostaglandin synthesis. Responses to electrical field stimulation following addition of the nitric oxide antagonist Nwo-nitro-L-arginine methyl ester (L-NAME) and apamin were evaluated. RESULTS: Electrical field stimulation (EFS) resulted in a reduction in the amplitude and frequency of contractile activity, followed by rebound contraction when EFS was stopped. Addition of L-NAME resulted in a significant reduction in inhibition of contractile activity. Addition of apamin also resulted in a significant reduction in inhibitory contractile activity at most stimulation frequencies. The combination of L-NAME and apamin resulted in a significant reduction in inhibition at all frequencies. CONCLUSION: Nitric oxide and a transmitter acting via an apamin-sensitive mechanism appear to be involved in inhibition of contractile activity of the third compartment in llamas. CLINICAL RELEVANCE: Results suggest that nitric oxide plays an important role in mediating contractile activity of the third compartment in llamas. Use of nitric oxide synthase inhibitors may have a role in the therapeutic management of llamas with lesions of the third compartment.

In vitro effects of nonsteroidal anti-inflammatory agents and prostaglandins I2, E2, and F2alpha on contractility of taenia of the large colon of horses.

OBJECTIVES: To determine the in vitro effect of various prostaglandins (PG) and nonsteroidal anti-inflammatory drugs (NSAID) on contractile activity of the large-colon taenia of horses. ANIMALS: 14 healthy horses. PROCEDURE: The taenia was collected from the ventral colon, cut into strips (2 X 10 mm), and mounted in a tissue bath system (20-ml capacity) that contained oxygenated Krebs buffer solution warmed to 37.5+/-0.5 C. After equilibration, incremental doses of PGE2, PGF2alpha, PGl2, flunixin meglumine, carprofen, ketoprofen, and phenylbutazone were added to the baths, and contractile activity was recorded. Magnitude of the response was calculated by comparing contractile activity before and after administration of the PG or NSAID to the tissue baths. RESULTS: PGE2 and PGF2alpha, caused a significant increase in contractile activity, whereas PGI2 induced an inhibitory response. Activity of NSAID on contraction was predominantly inhibitory. At low concentrations, ketoprofen induced an excitatory effect, which then became inhibitory at high concentrations. Compared with the other NSAID, carprofen significantly reduced contractile activity at lower concentrations. CONCLUSIONS: PGE2 and PGF2alpha appear to enhance contractility of large-colon taenia of horses, whereas PGI2 was inhibitory in the in vitro model. Administration of NSAID also inhibited contractility, with carprofen having the most potent effect. CLINICAL RELEVANCE: Administration of NSAID in combination with liberation of endogenous PG may predispose horses to development of intestinal stasis and subsequent impaction.

Effect of a leukocyte-depleting filter in an extracorporeal circuit used for low-flow ischemia and reperfusion of equine jejunum.

OBJECTIVE: To determine effect of leukocyte depletion on hematologic, morphologic, and metabolic variables of equine jejunum after induction of arterial low-flow ischemia and reperfusion by use of an extracorporeal circuit. ANIMALS: 14 healthy adult horses. PROCEDURE: A segment of jejunum was surgically removed and maintained in an isolated circuit for 3 hours (control group), arterial flow was reduced to 20% of baseline for 40 minutes followed by 1 hour of reperfusion (low-flow group), or leukocyte depletion was filter-induced, and low-flow ischemia and reperfusion were conducted as in the low-flow control group (filter-treated group). Various metabolic, hemodynamic, and histomorphologic variables were evaluated, including effects of electrical field stimulation and L-N-nitro-arginine-methyl-ester (L-NAME) on contractile activity. RESULTS: The extracorporeal circuit appeared to maintain the jejunum within physiologic limits for an extended period. Low-flow ischemia with reperfusion induced significant differences in various measurements, compared with control specimens. Significant differences were not detected between the low-flow and filter-treated groups. Myeloperoxidase activity was greater in the low-flow group than the control group, whereas a difference was not detected between control and filter-treated groups. CONCLUSIONS AND CLINICAL RELEVANCE: The extracorporeal circuit maintained intestine for 3 hours in a physiologic state and may be used for simulation of tissue injury. Leukocyte depletion generally did not attenuate the effects of low-flow ischemia and reperfusion on equine small intestine.

Use of an extracorporeal circuit to evaluate effects of ischemia and reperfusion of the equine large colon.

OBJECTIVE: To determine efficacy of an extracorporeal circuit to maintain a segment of equine large colon for 3.5 hours and to evaluate the effect of low arterial flow on histologic and metabolic variables. SAMPLE POPULATION: Segments of large colon from 15 healthy adult horses. PROCEDURE: The pelvic flexure was surgically removed and maintained in an isolated circuit. In the control group, tissue was evaluated for 3.5 hours, whereas in the low-flow group, arterial flow was reduced to 20% of baseline for 40 minutes followed by 2 hours of reperfusion. Various metabolic and hemodynamic variables were evaluated at 30-minute intervals. Effects of nitric oxide (NO) and L-N-nitro-arginine-methyl-ester (L-NAME) on contractile activity were determined, and histomorphologic evaluation was performed at the completion of the study. RESULTS: Low-flow ischemia with reperfusion caused significant histomorphologic differences, compared with the control group. In the low-flow group, significant differences included reduction in PaCO2, reduction in bicarbonate concentrations, increase in PaO2, and an increase in base deficit in arterial and venous blood samples. Other significant differences included increases in PCV, protein concentration, total WBC count, and albumin clearance for the low-flow group. Differences were not detected in inhibitory activity of the low-flow group relative to the control tissue with or without addition of NO and L-NAME. CONCLUSION: The extracorporeal circuit maintained a segment of equine intestine for 3.5 hours and can be used to simulate ischemic injury. The extracorporeal circuit provides the potential to investigate pharmaceutic agents that can minimize intestinal injury.

In vitro evaluation of a customized solution for use in attenuating effects of ischemia and reperfusion in the equine small intestine.

OBJECTIVE: To determine whether a customized solution could attenuate the effects of low-flow ischemia and reperfusion injury of the equine jejunum. SAMPLE POPULATION: A segment of jejunum obtained from 21 healthy adult horses. PROCEDURE: A segment of jejunum was maintained in an isolated extracorporeal circuit, and arterial flow was reduced to 20% of baseline for 40 minutes (ischemia) followed by 60 minutes of reperfusion. In 1 group, a customized solution was infused at a rate of 1 ml/min during low-flow ischemia and 3 ml/min during reperfusion. In a second group, the solution was infused at the same rate during low-flow ischemia, but it was infused at a rate of 7 ml/min during reperfusion. Control groups received lactated Ringer's solution administered at the same rates as for the customized solution. Various metabolic, hemodynamic, histologic, and permeability variables were recorded. RESULTS: A lower flow rate during reperfusion (3 ml/min) had a beneficial effect, compared with lactated Ringer's solution or the higher flow rate (7 ml/min). Use of the solution at this rate resulted in less histomorphologic injury and reduced mucosal permeability to albumin. CONCLUSIONS AND CLINICAL RELEVANCE: Use of a customized solution at a lower flow rate during repurfusion appeared to have a protective effect on equine jejunum when administered IV during low-flow ischemia and reperfusion.

Evaluation of nitric oxide as an inhibitory neurotransmitter in the equine ventral colon.

OBJECTIVE: To determine the role of nitric oxide and an apamin-sensitive nonadrenergic noncholingeric inhibitory transmitter on contractility of the ventral colon of horses. SAMPLE POPULATION: Strips of the circular and longitudinal muscle layers and taenia of the ventral colon from 14 horses. PROCEDURE: Muscle strips were suspended in tissue baths and attached to force transducers. Contractile activity of circular, longitudinal, and taenia muscle strips in response to electrical field stimulation was measured after addition of apamin and a nitric oxide inhibitor, N-nitro-L-arginine methyl ester (L-NAME). RESULTS: Electrical field stimulation reduced contractile activity in the circular muscle layer and taenia but not the longitudinal muscle layer. Addition of L-NAME significantly reduced inhibitory contractile activity at all frequencies for the circular muscle layer, whereas a significant effect was evident for the taenia only at the highest frequency. The combination of L-NAME and apamin resulted in a significant reduction in inhibition of the taenia at all frequencies but for circular muscle only at lower frequencies. CONCLUSIONS AND CLINICAL RELEVANCE: Nitric oxide and an apamin-sensitive neurotransmitter appear to mediate a component of inhibitory transmission in the circular muscle and taenia, but not the longitudinal muscle layer, of the equine ventral colon. Nitric oxide has a role in regulating contractile activity of the equine ventral colon, and nitric oxide synthase inhibitors may be useful in horses with ileus of the large colon.

Morphologic changes and xanthine oxidase activity in the equine jejunum during low flow ischemia and reperfusion.

OBJECTIVE: To determine whether xanthine oxidase and dehydrogenase activities are altered during low flow ischemia and reperfusion of the small intestine of horses. ANIMALS: 5 clinically normal horses without histories of abdominal problems. PROCEDURE: With the horse under general anesthesia, a laparotomy was performed and blood flow to a segment of the distal jejunum was reduced to 20% of baseline for 120 minutes and was then reperfused for 120 minutes. Biopsy specimens were obtained before, during, and after ischemia for determination of xanthine oxidase and dehydrogenase activities, and for histologic and morphometric analyses. RESULTS: Percentage of xanthine oxidase activity (as a percentage of xanthine oxidase and dehydrogenase activity) was not altered during ischemia and reperfusion. An inflammatory response developed and progressed during ischemia and reperfusion. Mucosal lesions increased in severity after ischemia and reperfusion. Mucosal surface area and volume decreased during ischemia and continued to decrease during reperfusion. Submucosal volume increased slightly during ischemia, and continued to increase during reperfusion. CONCLUSIONS AND CLINICAL RELEVANCE: Evidence for conversion of xanthine dehydrogenase to xanthine oxidase during ischemia was not found. Factors other than production of reactive oxygen metabolites may be responsible for progressive epithelial loss, decrease in mucosal surface area and volume, and increase in submucosal volume observed in this study. Other methods of determining xanthine oxidase activity that detect the enzyme in sloughed epithelial cells should be used to better define the importance of this pathway in jejunal reperfusion injury in horses.

Ultrastructural mucosal injury after experimental ischemia of the ascending colon in horses.

The ultrastructural injury that develops sequentially in the ascending colon during experimentally induced ischemia was examined in 6 halothane-anesthetized horses. Colonic ischemia was created by 2 types of vascular occlusion 24 cm proximal and distal to the pelvic flexure. In all horses, transmural vascular compression was created. The colonic venous circulation was obstructed in 3 horses, whereas in the other 3 horses, arterial and venous circulation was obstructed. Two additional horses were anesthetized as controls for determination of any morphologic alterations associated with the experimental protocol. Full-thickness colonic biopsy specimens were obtained from the antimesenteric border of the pelvic flexure at 0, 0.25, 0.5, 1, 1.5, 1.75, 2, 2.25, 2.5, 3, 3.5, 4, 4.5, and 5 hours during occlusion, and were studied by light and transmission electron microscopy. Morphologic alterations did not develop in the colon of control horses. Mucosal congestion was observed by light microscopy in the colon of horses with experimentally induced ischemia, but congestion developed early in those with obstructed colonic venous circulation, compared with those having arterial and venous obstruction. Inter- and intracellular vacuolation and loss of staining initially resulted in groups of 3 to 5 superficial luminal epithelial cells. Alterations in the glandular epithelium lagged behind those in the superficial epithelium, but were observed in both groups by 2 hours of obstruction. These changes progressed to 100% sloughing of all epithelium by 4.5 to 5 hours. The initial cellular alterations, which were observed by transmission electron microscopy, developed at 0.25 hour in horses with colonic venous obstruction and was characterized by inter- and intracellular edema.(ABSTRACT TRUNCATED AT 250 WORDS)

In vitro biomechanical comparison of the strength of the linea alba of the llama, using two suture patterns.

OBJECTIVE: To compare the strength of the sutured linea alba, in vitro, using 2 suture patterns. ANIMALS: 12 clinically normal llamas. PROCEDURE: 2 incisions in the linea alba of 12 llamas were closed with a simple continuous or inverted cruciate pattern, and tissue was harvested after 10 days. In 6 llamas the simple continuous line was intact; the inverted cruciate specimens contained 6 sutures. In 6 llamas, 1 knot was excised in the simple continuous pattern to simulate a failed line; the cruciate pattern contained 5 knots. Tissue sections were taken from cranial, between, and caudal to the linea alba incisions to compare fascial thickness. The sutured specimens were mounted in a mechanical testing system and tested to failure. A mixed-model ANOVA was used to evaluate the effects of suture pattern and incisional position on mechanical properties. RESULTS: Significant differences were not found between suture patterns or between location for yield force, failure force, or yield strain, whereas failure strain was lower for the intact simple continuous pattern than the inverted cruciate pattern (P = 0.003). From histomorphometric analysis, the caudal tissue specimens were significantly thinner than the middle tissue specimen cranial to the umbilicus (P = 0.006). CONCLUSION: There was no significant difference in monotonic breaking strength of the linea alba sutured with the simple continuous or inverted cruciate pattern. CLINICAL RELEVANCE: These results justify the use of the simple continuous pattern over the cruciate pattern for ventral midline closure in llamas because of the ease of placement and speed.

Microvascular permeability and endothelial cell morphology associated with low-flow ischemia/reperfusion injury in the equine jejunum.

Microvascular permeability of the jejunum of clinically normal equids and microvascular permeability associated with 60 minutes of ischemia (25% baseline blood flow) and subsequent reperfusion were investigated. Eight adult horses were randomly allotted to 2 equal groups: normal and ischemic/reperfusion injury. Lymphatic flow rates, mesenteric blood flow, and lymph and plasma protein concentrations were determined at 15-minute intervals throughout the study. Microvascular permeability was determined by estimates of the osmotic reflection coefficient, which was determined when the ratio of lymphatic protein to plasma protein concentration reached a constant minimal value as lymph flow rate increased (filtration-independent lymph flow rate), which occurred at venous pressure of 30 mm of Hg. Full-thickness jejunal biopsy specimens were obtained at the beginning and end of each experiment, and were prepared for light microscopy to estimate tissue volume (edema) and for transmission electron microscopy to evaluate capillary endothelial cell morphology. The osmotic reflection coefficient for normal equine jejunum was 0.19 +/- 0.06, and increased significantly (P < or = 0.0001) to 0.48 +/- 0.05 after the ischemia/reperfusion period. Microscopic evaluation revealed a significant increase (P < or = 0.0001) in submucosal and serosal volume and capillary endothelial cell damage in horses that underwent ischemia/reperfusion injury. Results indicate that ischemia/reperfusion of the equine jejunum caused a significant increase in microvascular permeability.