Proline and Glutamine Improve in vitro Callus Induction and Subsequent Shooting in Rice

This study was conducted to evaluate the effects of proline and glutamine on in vitro callus induction and subsequent regeneration and to develop a reproducible and highly efficient plant regeneration protocol in four rice genotypes, viz. Pawana, Jaya, Indrayani and Ambemohar. Considerable variation in response to plant growth regulators and amino acid supplements used was observed in all the four genotypes. Medium supplemented with proline and glutamine was shown to be superior to medium without proline and glutamine. The best callusing from mature embryo was observed on Murashige and Skoog(MS) medium supplemented with 2.0 mg/L 2,4-dichlorophenoxyacetic acid(2,4-D), 500 mg/L proline and 500 mg/L glutamine. Shoot induction was higher in the callus obtained from medium supplemented with 500 mg/L proline and 500 mg/L glutamine. The highest shoot regeneration frequency(83.2%) was observed on MS medium with 2.0 mg/L benzylaminopurine, 0.5 mg/L 1-naphthaleneacetic acid, 500 mg/L proline, and 500 mg/L glutamine in the callus obtained from MS medium supplemented with 2.0 mg/L 2,4-D, 500 mg/L proline and 500 mg/L glutamine. Among the four genotypes, Pawana has the highest regeneration efficiency(83.2%), whereas the regeneration efficiency of the rest three rice genotypes was in the range of 32.0% to 72.3%. This optimized regeneration protocol can be efficiently used for Agrobacterium mediated genetic transformation in rice.     

Glomerular filtration rate after tramadol, parecoxib and pindolol following anaesthesia and analgesia in comparison with morphine in dogs

ObjectiveTo compare the effects of morphine, parecoxib, tramadol and a combination of parecoxib, tramadol and pindolol on nociceptive thresholds in awake animals and their effect on glomerular filtration rate (GFR) in dogs subjected to 30 minutes of anesthesia.AnimalsEight adult mixed breed experimental dogs.Study designRandomized, controlled trial.MethodsDogs received 0.05 mg kg^?1 acepromazine subcutaneously (SC) as anaesthetic pre-medication. Thirty to sixty minutes later, they received either tramadol 3 mg kg^?1 intravenously, (IV), parecoxib (1 mg kg^?1 IV), a combination of tramadol 3 mg kg^?1 (IV), parecoxib 1 mg kg^?1 (IV) and pindolol 5 μg kg^?1 (SC), morphine (0.1 mg kg^?1 (IV) or 0.9% saline (2 mL). Anaesthesia was then induced with IV propofol to effect (2.9 ± 0.8 mg kg^?1) and maintained with halothane in oxygen for 30 minutes. Systolic arterial blood pressure was maintained above 90 mmHg with IV fluids and by adjusting the inspired halothane concentration. Post-treatment nociceptive thresholds to mechanical stimuli, expressed as percent of pre-treatment values, were compared between the treatments to assess the analgesic efficacy of the drugs. Plasma iohexol clearance (ICL), a measure of GFR, was estimated both before and 24 hours after induction of anaesthesia to study the drugs’ effects on renal perfusion. Nociceptive threshold and GFR data were compared using mixed model analysis in sas^®9.1.ResultsBoth tramadol and parecoxib produced similar analgesia, which was less than that of morphine. Their combination with pindolol produced analgesia comparable with morphine. None of the test drugs, either alone or in combination, reduced GFR.ConclusionTramadol and parecoxib (either alone or in combination) can increase nociceptive thresholds in awake dogs and have minimal effects on renal perfusion in normotensive dogs subjected to anaesthesia.     

Effects of tramadol on the minimum alveolar concentration of sevoflurane in dogs

ObjectiveTo evaluate the effect of tramadol on sevoflurane minimum alveolar concentration (MAC_SEVO) in dogs. It was hypothesized that tramadol would dose-dependently decrease MAC_SEVO.Study designRandomized crossover experimental study.AnimalsSix healthy, adult female mixed-breed dogs (24.2 ± 2.6 kg).MethodsEach dog was studied on two occasions with a 7-day washout period. Anesthesia was induced using sevoflurane delivered via a mask. Baseline MAC (MAC_B) was determined starting 45 minutes after tracheal intubation. A noxious stimulus (50 V, 50 Hz, 10 ms) was applied subcutaneously over the mid-humeral area. If purposeful movement occurred, the end-tidal sevoflurane was increased by 0.1%; otherwise, it was decreased by 0.1%, and the stimulus was re-applied after a 20-minute equilibration. After MAC_B determination, dogs randomly received a tramadol loading dose of either 1.5 mg kg^?1 followed by a continuous rate infusion (CRI) of 1.3 mg kg^?1 hour^?1 (T1) or 3 mg kg^?1 followed by a 2.6 mg kg^?1 hour^?1 CRI (T2). Post-treatment MAC determination (MAC_T) began 45 minutes after starting the CRI. Data were analyzed using a mixed model anova to determine the effect of treatment on percentage change in baseline MAC_SEVO (p < 0.05).ResultsThe MAC_B values were 1.80 ± 0.3 and 1.75 ± 0.2 for T1 and T2, respectively, and did not differ significantly. MAC_T decreased by 26 ± 8% for T1 and 36 ± 12% for T2. However, there was no statistically significant difference in the decrease between the two treatments.Conclusion and clinical relevanceTramadol significantly reduced MAC_SEVO but this was not dose dependent at the doses studied.     

Anesthetic management of an off-pump open-heart surgery in a dog

ObservationsA 9 year-old, 40 kg, female spayed Bouvier des Flandres was anesthetized for surgical removal of an intra-cardiac mass. Pre-anesthetic work-up included thoracic radiographs, which revealed moderate pleural effusion, and cardiac ultrasound, which identified a mass attached to the wall of the right ventricular outflow tract (RVOT). The mass caused dynamic obstruction of the RVOT during systole. The dog was pre-medicated with intravenous (IV) hydromorphone (0.05 mg kg^?1). Following pre-oxygenation, anesthesia was induced with ketamine (3.75 mg kg^?1, IV) and diazepam (0.18 mg kg^?1, IV). Anesthesia was maintained with isoflurane in oxygen, an intravenous constant rate infusion (CRI) of fentanyl (10–30 μg kg^?1 hour^?1) and a CRI of lidocaine (50–200 μg kg^?1 minute^?1). A right lateral thoracotomy was performed. The heart was stopped transiently with a cold cardioplegic solution for 7.83 minutes to allow the removal of the mass through an open-heart procedure. No cardiopulmonary bypass was used. The heart was successfully restarted after cardiopulmonary resuscitation with internal cardiac massage and internal defibrillation. The dog recovered uneventfully from anesthesia without any apparent neurological sequelae. Post-operative analgesia consisted of intercostal nerve blocks with bupivacaine, CRIs of fentanyl (2–5 μg kg^?1 hour^?1) and lidocaine (40 μg kg^?1 minute^?1) and with oral meloxicam (0.1 mg kg^?1). Five days following surgery, the dog was discharged from the hospital. Histopathology and immunohistochemistry of the mass identified an ectopic thyroid carcinoma.ConclusionsThis case showed the feasibility of whole body hypothermia and using a cold cardioplegic solution to induce cardiac arrest for a short open-heart procedure.     

Laparoscopic Mesh Incisional Hernioplasty in Five Horses

Objective— To report a technique for incisional hernioplasty in horses using laparoscopic placement of a prosthetic mesh.
Study Design— Case series.
Animals— Horses (n=5) with ventral median abdominal incisional hernia.
Methods— A telescope and 2 instrument portals were established bilaterally, lateral to and distant from the hernia margins. After exposure of the internal rectus sheath by removal of retroperitoneal fat with endoscopic scissors and monopolar cautery, a prosthetic mesh was introduced into the abdomen and secured intraperitoneally using transfascial sutures with or without supplemental endoscopic hernia fixation devices.
Results— Successful placement of the prostheses was achieved without major intra- or postoperative complications. Repairs were intact in all horses (follow-up range: 6–23 months) without evidence of adhesion formation. Cosmetic results compared favorably with those typically achieved using conventional, open hernioplasty techniques.
Conclusion— Incisional hernia repair in horses can be successfully achieved with a laparoscopic intraperitoneal mesh onlay technique.
Clinical Relevance— Laparoscopic mesh hernioplasty has promise as a safe and effective method for repair of incisional hernias in horses.     

Minimally Invasive Inguinal Approach for Tube Cystostomy

Objective— To report a technique for tube cystostomy placement via a minimally invasive inguinal approach and outcome in 9 dogs and 6 cats with urinary tract obstruction or detrusor atony.
Study Design— Case series.
Animals— Dogs (n=9) and cats (6).
Methods— Medical records (January 2004–January 2008) of dogs and cats that had tube cystostomy via an inguinal approach were reviewed. Retrieved data included signalment, diagnosis, surgical technique, and complications. Access to the bladder was through a muscle splitting approach in the inguinal region with the cystostomy tube placed through a skin incision made several centimeters proximal to this incision and secured in the bladder by a purse string suture. Cystopexy during closure of the muscle layers ensured secure closure and minimized the likelihood of uroabdomen if tube dislodgment occurred.
Results— Cystostomy tubes were placed in 5 cats as an emergency procedure for treatment of acute urinary tract obstruction or urethral rupture, and as an elective procedure in 9 dogs and 1 cat. No complications occurred during cystostomy tube placement. Postprocedural complications were minor (peristomal irritation in 2 dogs with latex catheters, catheter laceration, premature removal) and only occurred when tubes were retained for >4 weeks. Urinary tract infection at catheter removal in 6 dogs resolved with antibiotic administration.
Conclusions— An inguinal approach for cystostomy tube placement facilitated rapid catheter placement into the bladder with minimal soft tissue dissection. Cystopexy during abdominal wall closure provided peritoneal protection should premature dislodgement of the cystostomy tube occur.
Clinical Relevance— An inguinal approach should be considered for rapid tube cystostomy particularly in metabolically compromised animals.     

Biomechanical Comparison of Two Alternative Tibial Plateau Leveling Osteotomy Plates with the Original Standard in an Axially Loaded Gap Model: An In Vitro Study

Objective— To compare the axial compression stiffness of osteotomized canine tibiae stabilized with Slocum, Securos, or Synthes plates after a tibial plateau leveling osteotomy (TPLO) procedure. Study Design— In vitro, paired comparison of cadaveric tibial constructs subjected to mechanical testing under an axial load. Sample Population— Canine tibiae (n=16 pairs) from skeletally mature male and female dogs of various breeds (18–55 kg). Methods— Tibial pairs (n=16) were randomly assigned to 1 of 2 study cohorts (n=8 pairs/cohort): cohort 1, tibial osteotomy stabilization with a Slocum or a Securos plate, or cohort 2, tibial osteotomy stabilization with a Slocum or a Synthes plate. One tibia from each pair was stabilized with 1 of each plate design assigned to the cohort after TPLO. A 3.2 mm osteotomy gap was maintained during plate application in all constructs. Load and axial displacement were recorded while constructs were loaded to 2000 N in axial compression. Failure loads were not reported because no distinct yield point or failure point was evident within the load range for many specimens. Failure modes were recorded for each construct, and photographs of typical failures were obtained. Stiffness (N/mm) was calculated from load–displacement curves. Paired comparisons of mean stiffness were performed within study groups using a paired t‐test. Significance was set at P<.05. Results— The mean construct stiffnesses for the Slocum (383±183 N/mm) and Securos (258±64.1 N/mm) constructs were not significantly different (P=.164; power=0.566). The mean construct stiffness for the Synthes constructs (486±91.0 N/mm) was significantly greater than that of the Slocum constructs (400±117 N/mm); P=.0468. Modes of failure for the Slocum (16/16) and Securos (8/8) constructs included plastic deformation of the implant with valgus deformity combined with fibular luxation (2/16 Slocum; 1/8 Securos) or fibular fracture (2/16 Slocum; 4/8 Securos). Most Synthes constructs underwent elastic deformation (7/8). One Synthes construct fractured in the saggital plane through the tibial plateau depression at the point of load application. Conclusions— The Slocum and Securos plate/tibia construct have similar stiffness, whereas the Synthes/tibia constructs are significantly stiffer than the Slocum/tibia constructs. Modes of fixation failure observed in this model were consistent with TPLO fixation failures observed clinically. Clinical relevance— Construct stiffness in axial load varies with implant type. Implants that confer higher stiffness to the construct may result in greater fixation stability in tibial metaphyseal osteotomies.     

Effect of Prosthesis Number and Position on Rima Glottidis Area in Equine Laryngeal Specimens

Objective— To assess the effect of 1 or 2 laryngeal prosthetic sutures on rima glottidis areas in equine laryngeal specimens. Study Design— Experimental, randomized design. Animals— Cadaveric equine larynges (n=16). Methods— Larynges were collected from 10 horses; 2 sutures each were preplaced in the right and left sides of each larynx. A dorsal suture (DS) was placed through the caudal rim of the dorsal midline of the cricoid cartilage, under the cricopharyngeus muscle and through the proximal and rostral aspect of the muscular process. A lateral suture (LS) was placed 1.5 cm lateral to the DS and through the muscular process more distal and caudal to the 1st suture. Larynges were positioned in a customized stand and the rima glottidis photographed after each suture (LS or DS) or suture combination (CS) was tied in random sequence. An additional 6 larynxes were used to determine whether the tension applied to the sutures was repeatable. Sutures were preplaced in both the right and left side of each larynx as described above and each suture and CS was tied and released 3 times in each larynx. Photographs were taken of the rima glottidis after each suture or CS was tied generating 3 replicates for each suture configuration on each side of the 6 larynges. Results— Mean rima glottidis area was not different between DS and LS when tied alone (P=.85); however, mean area after CS (DS+LS) was greater than DS (P<.001) and LS (P<.001) alone. The coefficient of variation for the 6 suture patterns were low (1–7%) and the intraclass correlation coefficient estimates were very high (0.997–0.998) demonstrating excellent repeatability between replicates for each of the 3 suture configurations. Conclusion— Our results suggest that laryngoplasty using 2 prostheses; 1 placed dorsally in the cricoid and through the rostral and proximal muscular process and 1 placed 1.5 cm lateral to the 1st and more caudal and distal in the muscular process results in a greater cross sectional area of the rima glottidis than either suture used alone. Clinical Relevance— Seemingly prosthetic sutures contribute independently to each other in determining the contour of the rima glottidis. Use of 2 prosthetic sutures improves crosssectional area of the rima glottidis compared with each suture alone and may improve surgical outcome in laryngoplasty.     

Minimum Inhibitory Concentration and Postantibiotic Effect of Amikacin for Equine Isolates of Methicillin-Resistant Staphylococcus aureus In Vitro

Objective— To report the minimum inhibitory concentration (MIC) of amikacin sulfate for equine clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) and characterize the initial kill and duration of the postantibiotic effect (PAE) for selected strains.
Study Design— Experimental study.
Methods— Isolates of MRSA (n=35) had their amikacin MIC determined using the E-test agar diffusion method. Two isolates with MICs>256 μg/mL limit were further characterized using broth macrodilution. Six distinct isolates with amikacin MICs of 32, 48, 128 (2 isolates) and 500 (2 isolates) μg/mL had PAE determinations made over a range of amikacin concentrations from 31.25–1000 μg/mL using standard culture-based techniques.
Results— Median MIC of the 35 isolates was 32 μg/mL (range 2 to >256 μg/mL). Mean PAE of selected MRSA strains had an overall mean (all amikacin doses) of 3.43 hours (range 0.10–9.57 hours). PAE for MRSA exposed to amikacin at 1000 μg/mL was 6.18 hours (range 3.30–9.57 hours), significantly longer than that for all other concentrations ( P <.0001). There was no statistically significant effect of isolate MIC on PAE.
Conclusions— Isolates had a wide range of MIC; however, growth of all 6 selected strains were inhibited within the range of concentrations tested, including 2 strains with MICs of 500 μg/mL. PAE duration was not influenced by the MIC of amikacin but was significantly longer with treatment at 1000 μg/mL than at lower concentrations.
Clinical Relevance— Clinical isolates of MRSA are susceptible to amikacin at concentrations achieved by regional perfusion: however, the modest duration of PAE observed suggest that further laboratory and in vivo evaluation be conducted before recommending the technique for clinical use.     

Management of Tibial Fractures Using a Circular External Fixator in Two Calves

Objectives: To report the repair of tibial diaphyseal fractures in 2 calves using a circular external skeletal fixator (CEF). Study Design: Clinical report. Animals: Crossbred calves (n=2; age: 6 months; weight: 55 and 60 kg). Methods: Mid‐diaphyseal tibial fractures were repaired by the use of a 4‐ring CEF (made of aluminum rings with 2 mm K‐wires) alone in 1 calf and in combination with hemicerclage wiring in 1 calf. Results: Both calves had good weight bearing with moderate lameness postoperatively. Fracture healing occurred by day 60 in 1 calf and by day 30 in calf 2. The CEF was well maintained and tolerated by both calves through fracture healing. Joint mobility and limb usage improved gradually after CEF removal. Conclusions: CEF provided a stable fixation of tibial fractures and healing within 60 days and functional recovery within 90 days. Clinical Relevance: CEF can be safely and successfully used for the management of selected tibial fractures in calves.