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Objective-To evaluate the biological activity of dihydroartemisinin on canine osteosarcoma cell lines in vitro. Sample Population-4 canine osteosarcoma cell lines. Procedures-Cell viability assays were performed on canine osteosarcoma cell lines OSCA2, OSCA16, OSCA50, and D17 after 24, 48, and 72 hours of treatment with dihydroartemisinin at concentrations of 0.1 to 100muM. Apoptosis was assessed by use of an ELISA for free nuclosomal DNA fragmentation and by western blot analysis for cleavage of caspase 3. Cell cycle analysis was performed by use of staining with propidium iodide and flow cytometry. Detection of reactive oxygen species (ROS) was conducted in the D17 cell line by use of 6-carboxy-2′,7′-dihydrofluorescein diacetate and flow cytometry. Results-The concentration of dihydroartemisinin required for 50% inhibition of cell viability (IC(50)) was achieved in all 4 canine osteosarcoma cell lines and ranged from 8.7 to 43.6muM. Induction of apoptosis was evident as an increase in nucleosomal DNA fragmentation, cleavage of caspase 3, and an increase in the population in the sub G(0)/G(1) phase of the cell cycle detected by flow cytometry. Exposure to dihydroartemisinin also resulted in a decrease in the G(0)/G(1) population. Iron-dependent generation of ROS was detected in dihydroartemisinin-treated D17 cells; ROS generation increased in a dose-dependent manner. Conclusions and Clinical Relevance-Incubation with dihydroartemisinin resulted in biological activity against canine osteosarcoma cell lines, which included induction of apoptosis and arrest of the cell cycle. Clinical trials of dihydroartemisinin in dogs with osteosarcoma should be conducted.
Hosoya K Murahari S Laio A London CA Couto CG Kisseberth WC
Department of Veterinary Clinical Sciences, College of Veterinary Medicine, The Ohio State University, Columbus, OH 43210.
2008.69.1-4
Objective-To investigate sacroiliac luxation repair with positional screw insertion from the ventral surface of the sacral wing via a ventral abdominal approach in cats. Animals-18 European shorthair cats. Procedures-All cats underwent clinical examination including orthopedic and neurologic examination and assessment of lameness and pain before and immediately after surgery and 6 and 16 weeks after surgery. All sacroiliac luxations were stabilized with a single positional 2.4-mm cortical titanium self-tapping screw. The pelvic floor was also repaired in selected cats. Screw entry points and angles determined in a prior study of cadavers were used. Radiographs were taken before surgery and during follow-up evaluations to assess postoperative sacroiliac luxation reduction, implant placement, and repair stability. Results-All implants were placed correctly. Iatrogenic sciatic nerve injuries occurred in 2 cats. Median time to ambulation was 1.5 days for cats with sacroiliac luxation as the sole injury. Radiographic outcome of sacroiliac luxation repair was excellent in 15 of 17 repairs, good in 1 of 17 repairs, and poor in 1 of 17 repairs. Clinical outcome was excellent in 11 of 15 cats and good in 4 of 15 cats. Conclusions and Clinical Relevance-Insertion of a positional screw across the sacroiliac joint via a ventral abdominal approached can be an alternative to conventional techniques of sacroiliac luxation repair in cats. This novel technique allowed repair of bilateral sacroiliac luxation, repair of pelvic floor fractures, and treatment of soft tissue injuries of the abdominal cavity or abdominal organs with a single approach.
Borer LR Voss K Montavon PM
Department of Small Animal Surgery, Vetsuisse Faculty-Zurich, University of Zurich, Winterthurerstrasse 260, 8057 Zurich, Switzerland.
Objective-To investigate a technique for repair of sacroiliac luxation with positional screw insertion from the ventral surface of the sacral wing via a ventral abdominal approach. Sample Population-Hemipelvis specimens from cadavers of 5 small- to large-breed dogs and 9 European shorthair cats. Procedures-An optimal entry point and a safe drill corridor for implant placement were determined (4 hemipelvis specimens). Anatomic landmarks were identified, and the surgical technique for a ventral abdominal approach was described. Single positional screw placement was performed across the sacroiliac joint in 23 hemipelvis specimens. Screws were aimed at 25 degrees (n = 2), 35 degrees (2), and 45 degrees (19) angles to the vertical axis in a transverse plane (alpha angles) and at a 90 degrees angle to the longitudinal axis in a dorsal plane (beta angle). Implant placement was assessed by radiographic evaluation of the cadavers and of the hemipelvis specimens devoid of soft tissue. Results-By use of alpha angles of 35 degrees and 45 degrees , 20 of 21 implants were placed adequately; screws crossed the sacroiliac joint and penetrated the wing of the ilium without damaging adjacent nerves. The measured median alpha angle was 38 degrees , and the median beta angle was 88 degrees . One complication was recorded. Conclusions and Clinical Relevance-Cortical positional screw placement from the ventral aspect of the sacral wing by use of a ventral abdominal approach could be an alternative to conventional techniques. This novel technique may be useful for repair of bilateral sacroiliac luxation, treatment of concomitant soft tissue injuries of the caudal portion of the abdominal cavity or abdominal wall, and repair of pelvic floor fractures in a single approach.
Borer LR Voss K Montavon PM
Department of Small Animal Surgery, Vetsuisse Faculty-Zurich, University of Zurich, Winterthurerstrasse 260, 8057 Zurich, Switzerland.
Objective-To evaluate the use of endoscopy in conjunction with a gastropexy technique in dogs as a potential means to aid prevention of gastric dilatation-volvulus. Animals-12 healthy adult medium- and large-breed dogs. Procedures-12 adult research dogs that had no abnormal physical examination findings each underwent an endoscopically assisted gastropexy procedure. On completion of the procedure, the dogs were euthanized and exploratory laparotomies were performed to evaluate the surgical site. Data recorded included anatomic location of the gastropexy, gastropexy length, and duration of procedure as well as any complications. Results-Mean +/- SD gastropexy length was 3.3 +/- 0.25 cm, and mean duration of surgery was 18 +/- 7 minutes. In each dog, the stomach was located in its normal anatomic position and all gastropexies were sutured to the abdominal wall at the level of the pyloric antrum. The only complications during the procedure were needle bending and breakage at the time of stay suture placement. Conclusions and Clinical Relevance-On the basis of these findings, it appears that endoscopically assisted gastropexy is a simple, fast, safe, and reliable method of performing a prophylactic gastropexy in dogs when undertaken by a person who is skilled in endoscopy. Such a procedure maximizes the benefits of decreased morbidity and shorter duration of anesthesia associated with minimally invasive surgery. Further clinical studies are warranted to evaluate the long-term efficacy of this procedure in dogs at risk for development of gastric dilatation-volvulus.
Dujowich M Reimer SB
Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Iowa State University, Ames, IA 50011-1250.
Objective-To characterize the pharmacokinetics of remifentanil in conscious cats and cats anesthetized with isoflurane. Animals-6 cats. Procedures-Remifentanil (1 mug/kg/min for 5 minutes) was administered IV in conscious cats or cats anesthetized with 1.63% isoflurane in oxygen in a randomized crossover design. Blood samples were obtained immediately prior to remifentanil administration and every minute for 10 minutes, every 2 minutes for 10 minutes, and every 5 minutes for 10 minutes after the beginning of the infusion. Blood was immediately transferred to tubes containing citric acid, flash frozen in liquid nitrogen, and stored at -80 degrees C until analysis. Blood remifentanil concentration was determined by use of liquid chromatography-mass spectrometry. Remifentanil concentration-time data were fitted to compartment models. Results-A 2-compartment model (with zero-order input because of study design) best described the disposition of remifentanil in awake and isoflurane-anesthetized cats. The apparent volume of distribution of the central compartment, the apparent volume of distribution at steady state, the clearance, and the terminal half-life (median [range]) were 1,596 (1,164 to 2,111) and 567 (278 to 641) mL/kg, 7,632 (2,284 to 76,039) and 1,651 (446 to 29,229) mL/kg, 766 (408 to 1,473) and 371 (197 to 472) mL/min/kg, and 17.4 (5.5 to 920.3) and 15.7 (3.8 to 410.3) minutes in conscious and anesthetized cats, respectively. Conclusions and Clinical Relevance-The disposition of remifentanil in cats was characterized by a high clearance. Isoflurane anesthesia significantly decreased the volume of the central compartment, likely by decreasing blood flow to vessel-rich organs.
Pypendop BH Brosnan RJ Siao KT Stanley SD
Department of Surgical and Radiological Sciences, School of Veterinary Medicine, University of California, Davis, CA 95616.
Objective-To determine the effects of body position on intraocular pressure (IOP) in dogs without glaucoma. Animals-24 healthy dogs with no evidence of glaucoma. Procedures-Dogs underwent ophthalmic examinations to ensure that no IOP-affecting ocular diseases were present. Each dog was sequentially placed in dorsal recumbency, sternal recumbency, and sitting position. For each of the 3 positions, IOP in the right eye was measured by use of an applanation tonometer immediately after positioning (0 minutes) and after 3 and 5 minutes had elapsed. The initial body position was randomly assigned; each position followed the other positions an equal number of times, and IOP measurements were initiated immediately after moving from one body position to the next. Proparacaine hydrochloride (0.5%) was applied to the right eye immediately prior to IOP measurements. Results-Intraocular pressure was affected by body position. During the 5-minute examination, IOP decreased significantly in dogs that were dorsally recumbent or sitting but did not change significantly in dogs that were sternally recumbent. For the 3 positions, overall mean IOP differed significantly at each time point (0, 3, and 5 minutes). Mean IOP in dorsal recumbency was significantly higher than that in sternal recumbency at 0 and at 3 minutes; although the former was also higher than that in sitting position at 3 minutes, that difference was not significant. Conclusions and Clinical Relevance-Body position affects IOP in dogs. When IOP is measured in dogs, body position should be recorded and consistent among repeat evaluations.
Broadwater JJ Schorling JJ Herring IP Elvinger F
Department of Small Animal Clinical Sciences, Virginia-Maryland Regional College of Veterinary Medicine, Blacksburg, VA 24061.
Objective-To determine the effects of intestinal ischemia and reperfusion on the expression of tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 mRNAs in the jejunum, liver, and lungs of dogs. Animals-8 healthy adult Beagles. Procedures-In each dog, the cranial mesenteric artery was occluded for 0 (control group; n = 4) or 60 (I-R group; 4) minutes, followed by reperfusion for 480 minutes; serum TNF-alpha and IL-6 activities and expression levels of TNF-alpha and IL-6 mRNAs in jejunal, hepatic, and lung tissues were measured before and at the end of the ischemic period and at intervals during reperfusion. For each variable, values were compared between the control and I-R groups at each time point. Results-Compared with the control group, serum IL-6 activity increased significantly after 180 minutes of reperfusion in the I-R group; also, jejunal TNF-alpha mRNA expression increased significantly after 60 (peak) and 180 minutes of reperfusion. In the I-R group, expressions of IL-6 mRNA in the liver and TNF-alpha and IL-6 mRNAs in the lungs increased significantly at 480 minutes of reperfusion, compared with the control group. Serum TNF-alpha activity, expression of IL-6 mRNA in the jejunum, and expression of TNF-alpha mRNA in the liver in the control and I-R groups did not differ. Conclusions and Clinical Relevance-Results indicated that the liver, lungs, and jejunum contributed to the production of TNF-alpha and IL-6 after intestinal ischemia and reperfusion in dogs, suggesting that intestinal ischemia and reperfusion induce a systemic proinflammatory cytokine response in dogs.
Nezu Y Nezu Y Shigihara K Harada Y Yogo T Hara Y Tagawa M
Division of Veterinary Surgery, Nippon Veterinary and Life Science University, 1-7-1 Kyonan-cho, Musashino-shi, Tokyo, 180-8602 Japan.
The promise of cellular therapy lies in the repair of damaged organs and tissues in vivo as well as generating tissue constructs in vitro for subsequent transplantation. Unfortunately, the lack of available donor cell sources limits its ultimate clinical applicability. Stem cells are a natural choice for cell therapy due to their pluripotent nature and self-renewal capacity. Creating reserves of undifferentiated stem cells and subsequently driving their differentiation to a lineage of choice in an efficient and scalable manner is critical for the ultimate clinical success of cellular therapeutics. In recent years, a variety of biomaterials have been incorporated in stem cell cultures, primarily to provide a conducive microenvironment for their growth and differentiation and to ultimately mimic the stem cell niche. In this review, we examine applications of natural and synthetic materials, their modifications as well as various culture conditions for maintenance and lineage-specific differentiation of embryonic and adult stem cells.
Eileen Dawsona Gazell Mapilia Kathryn Ericksona Sabia Taqvia Krishnendu RoyaEmail:kroy@mail.utexas.edu
[a]Department of Biomedical Engineering, The University of Texas at Austin, Austin, TX 78712, USA
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Objective-To investigate the effects of sterile fine dust aerosol inhalation on antibody responses and lung tissue changes induced by Mucor ramosissimus or Trichoderma viride spores following intratracheal inoculation in goats. Animals-36 weanling Boer-Spanish goats. Procedures-6 goats were allocated to each of 2 M ramosissimus-inoculated groups, 2 T viride-inoculated groups, and 2 control (tent or pen) groups. One of each pair of sporetreated groups and the tent control group were exposed 7 times to sterilized fine feedyard dust (mean +/- SD particle diameter,<7.72 +/- 0.69 mum) for 4 hours in a specially constructed tent. Goats in the 4 fungal treatment groups were inoculated intratracheally 5 times with a fungal spore preparation (30 mL), whereas tent control goats were intratracheally inoculated with physiologic saline (0.9% NaCl) solution (30 mL). Pen control goats were not inoculated or exposed to dust. Goats received an IV challenge with equine RBCs to assess antibody responses to foreign antigens. Postmortem examinations were performed at study completion (day 68) to evaluate lung tissue lesions. Results-5 of 7 deaths occurred between days 18 and 45 and were attributed to fine dust exposures prior to fungal treatments. Fine dust inhalation induced similar lung lesions and precipitating antibodies among spore-treated goats. Following spore inoculations, dust-exposed goats had significantly more spores per gram of consolidated lung tissue than did their nonexposed counterparts. Conclusions and Clinical Relevance-Fine dust inhalation appeared to decrease the ability of goats to successfully clear fungal spores from the lungs following intratracheal inoculation.
Purdy CW Layton RC Straus DC Ayers JR
USDA, Agricultural Research Service, Conservation and Production Research Laboratory, PO Drawer 10, Bushland, TX 79012.
Objective-To examine whether in vitro treatment with trans-10, cis-12 conjugated linoleic acid (t10c12-CLA) restores the phagocytic capacity and oxidative burst activity (OBA) of canine polymorphonuclear neutrophilic leukocytes (PMNs) exposed to methylprednisolone sodium succinate (MPSS). Sample Population-Peripheral blood PMNs obtained from 12 healthy Beagles. Procedures-The experimental design involved administration of a high dose of MPSS, which is the recommended protocol for dogs with acute spinal cord injury. To evaluate PMN function, blood samples were collected from dogs before IV injections of doses of MPSS or saline (0.9% NaCl) solution (time 0) and 2, 12, and 24 hours after injections ceased. Polymorphonuclear neutrophilic leukocytes were isolated from blood samples and incubated with t10c12-CLA alone or t10c12-CLA in combination with N-acetylcysteine (an antioxidant agent). Phagocytic capacity and OBA were measured simultaneously by use of flow cytometry. Results-The phagocytic capacity and OBA of PMNs were suppressed by IV injection of MPSS and restored 12 hours after injection ceased. In vitro treatment with t10c12-CLA enhanced the phagocytic capacity and OBA of PMNs, regardless of whether dogs had been treated with MPSS. Effects of t10c12-CLA on OBA were detected only when phagocytosis was stimulated by microspheres. Use of N-acetylcysteine attenuated the stimulatory effects of t10c12-CLA. Conclusions and Clinical Relevance-Exposure to t10c12-CLA enhanced the phagocytic capacity and OBA of canine PMNs, and this effect may have involved t10c12-CLA-induced generation of reactive oxygen species.
Kang JH Yang MP
Laboratory of Veterinary Internal Medicine, Department of Veterinary Medicine, College of Veterinary Medicine, Chungbuk National University, Cheongju, Chungbuk 361-763, Republic of Korea.