Fractionated oral dosing and its effect on cyclophosphamide pharmacokinetics in dogs with high-grade multicentric lymphoma

Cyclophosphamide (CP) is an alkylating agent commonly included in multi-drug treatment protocols for canine cancer. As a prodrug, CP requires hepatic metabolism for activation to the intermediate compound 4-hydroxycyclophosphamide (4-OHCP) which then spontaneously forms alkylating phosphoramide mustard. CP is frequently administered in a fractionated manner, with the total dose given over multiple days. CP is reported to cause auto-induction of metabolism in humans, with faster CP clearance and relatively increased 4-OHCP formation following fractionated versus bolus dosing, however canine pharmacokinetic studies of CP dose fractionation are lacking. The study objective was to evaluate the pharmacokinetics of fractionated oral CP dosing at a dose of 200–250 mg/m² over 3 to 4 days in a prospectively identified population of cancer-bearing dogs. Plasma concentrations of CP and 4-OHCP were measured by ultra-high performance liquid chromatography tandem-mass spectrometry in eight dogs following the first and last doses to assess for auto-induction of CP metabolism. No significant difference in the rate of CP elimination between first and last doses were detected (0.73 ± 0.46 vs. 1.22 ± 0.5 h⁻¹; p = .125). Additionally, no significant difference in dose-normalized 4-OHCP exposure was identified between first and last doses (5.9 ± 2.1 vs. 7.9 ± 6.4 h × ng/ml; p = .936). These results suggest that fractionated dosing may not increase exposure to the active metabolite of CP in dogs as it does in humans. As such, standard bolus dosing and fractionated dosing may be equivalent in terms of bio-activation of CP in dogs administered a dose of 200–250 mg/m².     

Tracking change in a human‐dominated landscape: developing conservation guidelines using freshwater fish

• 1. To conserve biodiversity in a human‐dominated landscape, a science‐based inventory and monitoring plan is needed that quantifies existing resources, isolates drivers that maintain natural communities, determines harmful stressors, and links ecological drivers and human stressors. A tactical approach is proposed for conservation planning using freshwater fish at the Cape Cod National Seashore.
• 2. Freshwater fish are well studied and occur across environmental gradients. The lentic systems at the national park are relatively pristine yet are enveloped by a region of high population density. Using fish community data, three steps were taken for tracking anthropogenic impacts in a human dominated landscape. First, fish and potential drivers were sampled intensively along a gradient to determine which fish metrics reflect natural communities and which abiotic and biotic factors structure them. Second, emerging and existing regional human threats were identified. Third, these human threats were linked to the potential drivers that maintain natural communities to identify the most informative metrics to monitor and track change.
• 3. Fish communities, water quality, habitat, and food resources were sampled concurrently in 18 ponds in 1999 and 2000. Three common fish species explained 98% of variation in numbers across systems. Based on ecological relationships, pH, depth, vegetation, prey, and community complexity were determined to maintain biodiversity of freshwater fish communities.
• 4. The primary human threats here included: development‐related, land‐use changes; non‐point source pollution; eutrophication from septic systems; and introduced species that are a byproduct of high human visitation. These are common threats in many rapidly urbanized areas and are likely to have relevance to many sites.
• 5. To track the impact of emerging threats to freshwater ponds related to increased human population, monitoring changes in water quality, vegetated habitat, fish diversity, and trophic interactions are recommended.

Copyright © 2008 John Wiley & Sons, Ltd.     

Compositional and moisture content effects on the biodegradability of zein/ethylcellulose films

The effect of moisture content and film composition on biodegradability is the focus of this study. Flexible films were first characterized for the effect on water sorption isotherms of relative humidity, temperature, zein content, and the addition of the plasticizers stearic acid, poly(ethylene glycol), or etoxylated ricine oil. Zein/ethylcellulose (EC) mixture films had a behavior between that for pure zein and EC films, which had the lowest water sorption. For films with plasticizer, the lowest water sorption at 25 degrees C was observed for those with stearic acid. Biodegradability of zein/EC films, evaluated using bacterial cultures selected for their zein proteolytic activity and isolated from a local solid waste landfill and a lagoon, showed no plasticizer effect even though its effect on moisture content was significant. Large differences were observed at different film zein concentration with the highest biodegradability for 100% zein. However, biodegradability did not mimic the water sorption behavior of zein/EC mixture films.     

Safety of a modified-live combination vaccine against respiratory and reproductive diseases in pregnant cows

A combination vaccine (Bovi-Shield FP4 + L5, Pfizer Animal Health) containing modified-live virus (MLV) components against bovine herpesvirus-1 (BHV-1), bovine viral diarrhea virus BVDV), parainfluenza virus-3 (PI3), bovine respiratory syncytial virus (BRSV), and inactivated cultures of Leptospira canicola, grippotyphosa, hardjo, icterohaemorrhagiae, and pomona was evaluated for safety in pregnant beef and dairy animals. Heifers vaccinated prebreeding with the minimum immunizing dose (lowest antigen level initiating immunizing effects) of the vaccine's MLV BHV-1 or BVDV components and during pregnancy (approximately 200 days of gestation) with vaccine containing 10x doses of the same BHV-1 and BVDV components delivered live, healthy calves that were determined to be serologically negative (titer less than 1:2) for neutralizing antibodies to BHV-1 and BVDV prior to nursing. Additionally, in three field safety studies, previously vaccinated cows and heifers that received a field dose (vaccine containing antigen levels required for commercial sale of the MLV combination vaccine during either the first, second, or third trimester of pregnancy had abortion rates similar to those of pregnant cows and heifers vaccinated during the same stage of pregnancy with sterile water diluent.     

Comparison of radiographic assessments of the tibial plateau slope in dogs

OBJECTIVES: To evaluate the accuracy of 2 radiographic methods used to assess tibial plateau slope (TPS) in dogs and evaluate effects of film digitization and radiographic beam placement on TPS measurements. SAMPLE POPULATION: 16 hind limbs from dog cadavers weighing > 20 kg. PROCEDURES: Radiographs of tibiae were made with the radiographic beam centered over the stifle joint and midshaft of the tibia. Tibiae were collected, the femorotibial contact area was determined, and slope of the medial tibial condyle in relation to the tibial shaft was measured. Radiographs were digitized. Slope of the medial tibial condyle was measured on printed and digitized radiographs read in random order by 6 examiners unaware of anatomic measurements. Three examiners used a conventional measuring technique, and 3 examiners used an alternative measuring technique. RESULTS: Anatomic measurements were significantly higher than radiographic measurements made by use of the conventional interpretation method but did not differ from radiographic measurements made by use of the alternate method. Measurements from printed radiographs were lower than measurements from digitized radiographs for the 4 most experienced examiners. CONCLUSIONS AND CLINICAL RELEVANCE: Measurements made by use of a line tangential to the cranial, linear portion of the medial tibial condyle at the femorotibial contact point were accurate measurements of the anatomic TPS. Measurements made by use of the conventional TPS measurement method underestimated the anatomic TPS. Measurements made on digitized radiographs were typically more accurate than measurements made on printed radiographs.     

Demographic, clinical, and radiographic features of bronchiectasis in dogs: 316 cases (1988-2000)

OBJECTIVE: To determine demographic, clinical, and radiographic features of bronchiectasis in dogs. DESIGN: Retrospective study. ANIMALS: 289 dogs identified through the Veterinary Medical Database (VMDB) and 27 dogs examined at the North Carolina State University Veterinary Teaching Hospital. PROCEDURE: Demographic characteristics of dogs identified through the VMDB were compared with characteristics of the entire population of dogs entered in the VMDB. Medical records of dogs examined at the teaching hospital were reviewed; the diagnosis was confirmed through review of thoracic radiographs. RESULTS: Analysis of data from the VMDB indicated that American Cocker Spaniels, West Highland White Terriers, Miniature Poodles, Siberian Huskies, English Springer Spaniels, and dogs > 10 years old had an increased risk of bronchiectasis. Among dogs examined at the teaching hospital, coughing was the most common clinical sign. There was evidence for excessive airway mucus but not hemorrhage. A variety of bacterial organisms were isolated from tracheal wash and bronchoalveolar lavage samples. On thoracic radiographs, cylindrical bronchiectasis, generalized disease, and right cranial lung lobe involvement were most common. Seven of 14 dogs for which follow-up radiographs were available did not have any progression of radiographic lesions. Median duration of clinical signs prior to diagnosis of bronchiectasis was 9 months (range, 1 day to 10 years). Median survival time was 16 months (range, 2 days to 72 months). CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that despite substantial clinical abnormalities, dogs with bronchiectasis may survive for years. Certain purebred dogs and older dogs may have an increased risk of developing bronchiectasis.     

Real-time polymerase chain reaction testing for the detection of Mycobacterium genavense and Mycobacterium avium complex species in avian samples

Diagnosis of avian mycobacteriosis, caused by Mycobacterium genavense or species belonging to the Mycobacterium avium complex (MAC), is problematic. Polymerase chain reaction (PCR) offers rapid and sensitive detection of minute quantities of DNA, and conventional protocols have been used for evaluating avian specimens. The recent development of real-time PCR offers several advantages over conventional PCR. In attempts to improve diagnosing avian mycobacteriosis, a real-time TaqMan PCR assay was developed targeting the 65-kD heat shock protein gene of M. genavense and MAC spp. Nineteen reference isolates, 16 clinical isolates, and 32 avian tissue samples were used to evaluate the assay. When sufficient amplicons were produced, the species of mycobacteria was determined by standard sequencing of TaqMan PCR products and compared with results from commercial mycobacteriology laboratories and/or standard sequencing of conventional PCR products. The TaqMan PCR detected DNA from reference isolates of M. genavense, MAC spp., and Mycobacterium tuberculosis complex spp. Of the clinical isolates, the TaqMan PCR detected DNA from 10 of 12 Mycobacterium avium avium isolates and two of three Mycobacterium avium intracellulare isolates. For the tissue samples, the TaqMan PCR amplified DNA in six of nine samples that were identified by sequencing of conventional PCR products and/or by commercial mycobacteriology laboratories as being MAC spp. positive and three of four samples that were positive for M. genavense. There was some disagreement between speciation results from the TaqMan PCR and those from commercial mycobacteriology laboratories or conventional PCR or both. This disagreement was suspected to be because of relatively small numbers of base pairs in the TaqMan PCR products. The TaqMan PCR may provide a useful tool for evaluating clinical samples for DNA from mycobacteria species that most commonly infect birds; however, further refinement is needed in order to improve sensitivity and provide more accurate speciation.     

Use of the distomedial-proximolateral oblique radiographic view of the elbow joint for examination of the medial coronoid process in dogs

OBJECTIVE: To describe and evaluate a new radiographic view of the elbow joint in dogs that would potentially enhance observation of the medial coronoid process (MCP). SAMPLE POPULATION: 20 cadaver limbs from 10 dogs and clinical examination of 100 elbow joints of 53 dogs. PROCEDURE: Twenty elbow joints from 10 cadavers were imaged by use of mediolateral, flexed mediolateral, craniocaudal, craniolateral-caudomedial oblique (Cr15L-CdMO), and distomedial-proximolateral oblique (Di35M-PrLO) radiographic views before and after placement of 3 lead pellets placed on the cranial, medial, and craniodistal aspect of the MCP. Three examiners independently reviewed these radiographs. One hundred elbow joints of 53 dogs with forelimb lameness and signs of pain elicited on palpation of the elbow joint were examined. These joints were radiographed and treated by use of arthroscopy. Three examiners independently graded the radiographs. RESULTS: The MCP was identified on all Di35M-PrLO views made during the anatomic study. The Di35M-PrLO view had the largest area under the receiving operating characteristic (ROC) curve for detection of abnormalities of the MCP. Fractured and nonfractured MCP could only be significantly differentiated on Di35M-PrLO and mediolateral views. The Di35M-PrLO view had a higher agreement between examiners than other radiographic views for detection of fractures of the MCP. CONCLUSION AND CLINICAL RELEVANCE: The Di35M-PrLO view enhances the identification of anomalies and fragmentation of the MCP in dogs, compared with other radiographic views. The Di35M-PrLO view may be of benefit for early screening of dogs potentially affected with elbow dysplasia.     

Protein microanalysis of animal tears

Sub-microlitre volumes of normal koala, mouse, dog, rat and cat tears were fractionated using size exclusion-high performance liquid chromatography (SE - HPLC), giving reproducible profiles which were different for each species. Microlitre volumes of tears were also fractionated using sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS - PAGE), resulting in good separation of individual tear proteins with a species specific distribution. Tears from koalas with conjunctivitis and mice with keratitis were similarly examined and showed mostly quantitative changes. These simple, rapid techniques gave reproducible results and, in contrast to conventional separation techniques, used easily obtainable volumes (as little as 0.75 microl) of tears. Their expansion could allow isola tion, identification and quantitation of individual tear components, enabling effective investigation of changes occurring in disease.