Enhancement of bone healing using non-glycosylated rhBMP-2 released from a fibrin matrix in dogs and cats

OBJECTIVES: To test a non-glycosylated recombinant human bone morphogenetic protein-2 (ngly-rhBMP-2)/fibrin composite, which has been shown experimentally to enhance healing of bone defects in rodents, in a clinical case series of dogs and cats undergoing treatment for fracture non-unions and arthrodesis. METHODS: A ngly-rhBMP-2/fibrin composite was applied in 41 sites in 38 dogs and cats for which a cancellous bone autograft was indicated, replacing the graft. RESULTS: Bridging of the bone defect with functional bone healing was achieved in 90 per cent of the arthrodesis and fracture nonunions treated in this manner. CLINICAL SIGNIFICANCE: This prospective clinical study demonstrates the beneficial effects of ngly-rhBMP-2 in a specially designed fibrin matrix on the treatment of bone defects, and validates the use of this composite as an alternative to bone autografts in dogs and cats.     

Colorectal polyps of the dog: a clinicopathologic study of 17 cases.

Seventeen cases of colorectal polyps in the dog were seen at the Ontario Veterinary College over a 12-year period. The mean age of occurrence was 6.9 years, and males and females were almost equally affected. The Collie was the most frequently affected breed (4/17). The clinical signs were typically dyschezia, periodic intermittent diarrhea and melena, and rectal prolapse of the polyp. The histopatholic classifcation were hyperplastic polyp (1), papillary adenoma (1), tubular adenoma (4), papillotubular adenoma (10), and unclassified (1). Severe epithelial atypia, likely carcinoma in situ, was apparent in 5 of the papillotubular adenomas. Retrospective analysis of survival data after polypectomy suggested a good prognosis, but the results were inconclusive and it is possible that some of the polyps represented premalignant lesions. Large polyps (greater than 1 cm in diameter) frequently had marked epithelial atypia and were considered more likely to recur or to become malignant.     

The non-diseased reactor: considerations on the interpretation of screening test results

The mathematical relationships between the sensitivity and specificity of screening tests and the proportion of animals which are positive to the test, but which have no visible lesions (NVL) at subsequent examination or necropsy, are outlined briefly. NVL reactors may be defined as a proportion of the total number of reactors to the test (reactor NVL per cent) or of the total number of animals tested (herd NVL per cent). Neither the herd nor reactor NVL per cent can provide valid estimates of the efficacy of any particular test, unless the precise prevalence of disease in the tested herd is determined. However under conditions of low or moderate prevalence specificity is approximately equal in magnitude to 100-herd NVL per cent but sensitivity cannot be reliably estimated from either the herd or reactor NVL per cent.     

On the weight-bearing function of the medial coronoid process in dogs

The shape of and proportions between the surface areas of the medial coronoid process (MCP) and the fovea of the radial head were determined in 88 juvenile dogs and 146 adult dogs grouped as giant, large, mid-sized, chondrodystrophic, or small dogs. Thereby, the longitudinal (length) and transverse (width) extension of the MCP and fovea of the radial head have been measured. Original values were used to describe changes of the parameters attributed to growth. Normalized values (i.e. values expected in case of a width of the fovea of the radial head of 20 mm) were used to determine potential differences between constitutional types. All original values increased during growth (P < 0.05) except for the width and length of the MCP in chondrodystrophic and small breeds. Normalized values revealed a proportional decrease in width and length of the MCP during growth (P < 0.05) compared with the radial head. In adults, the normalized MCP was widest in giant dogs followed by large, mid-sized, small, and chondrodystrophic breeds. The MCP was also longest in giant dogs but shortest in large and chondrodystrophic dogs with those of large dogs being significantly (P < 0.05) shorter than those in giant, mid-sized and small dogs. Present results suggest that a deficiency in length-growth of the MCP--which has been present especially in large dogs--results in smaller humeral contact areas and decreased weight-bearing capacity of the MCP. Because loading forces acting on the MCP increase with body weight, the condition noted in large dogs might increase the risk of fragmentation of the MCP in these.     

Oil concentration and fatty-acid profile of naturalized <Emphasis Type="Italic">Helianthus annuus</Emphasis> populations from Australia

Although wild annual sunflower (Helianthus annuus L.) is indigenous to North America, it has been both inadvertently and intentionally introduced into other countries, such as Australia, where it has become naturalized. Prior to this research, there were no collections of naturalized H. annuus from Australia in the USDA National Plant Germplasm System gene bank. The possibility exists that naturalized Australian populations may contain distinct traits, such as oil content and fatty-acid composition, differing from their North American progenitors due to the different environments, diseases, and insect-pest complexes. The objective of this study was to collect naturalized H. annuus populations from the five states of Australia and to analyze them for oil content and fatty-acid composition. Fifty-six populations were collected throughout the broad distributional range of this species and then analyzed for oil content and fatty-acid composition. The mean oil content of the H. annuus populations was 28%, ranging from a low of 19% to a high of 31%, similar to populations of this species in its native North American habitats. Fatty-acid profiles were also similar for North American wild populations, Australian naturalized populations, and improved cultivars. Oil concentration of hybrids can be rapidly increased to acceptable levels by backcrossing with cultivated sunflower. Based on this fact, there should be little concern about the relatively low oil content of these naturalized populations when they are used as a source of other unique traits for cultivated sunflower improvement.     

Functional and structural assessment of retinal sheet allograft transplantation in feline hereditary retinal degeneration

Purpose  To investigate whether sheets of fetal retinal allografts can integrate into the dystrophic Abyssinian cat retina with progressive rod cone degeneration.
Methods  Fetal retinal sheets (cat gestational day 42), incubated with BDNF microspheres, were transplanted to the subretinal space of four cats at an early disease stage. Cats were studied by fundus examinations, bilateral full-field flash ERGs, and indocyanine green and fluorescein angiograms up to 4 months following surgery. E42 donor and transplanted eyes were analyzed by histology and immunohistochemistry for retinal markers.
Results  Funduscopy and angiography showed good integration of the transplants in two of four cats, including extension of host blood vessels into the transplant and some scarring in the host. In these two, transplants were found in the subretinal space with laminated areas, with photoreceptor outer segments in normal contacts with the host retinal pigment epithelium. In some areas, transplants appeared to be well-integrated within the host neural retina. Neither of these two cats showed functional improvement in ERGs. In the other two cats, only remnants of donor tissue were left. Transplants stained for all investigated cellular markers. No PKC immunoreactivity was detected in the fetal donor retina at E42, but developed in the 4-month-old grafts.
Conclusions  Fetal sheet transplants can integrate well within a degenerating cat retina and develop good lamination of photoreceptors. Functional improvement was not demonstrated by ERG in cats with well-laminated grafts. Transplants need to be further evaluated in cat host retinas with a more advanced retinal degeneration using longer follow-up times.     

Short-term impacts of nutrient manipulations on leaf gas exchange and biomass partitioning in contrasting 2-year-old Pinus taeda clones during seedling establishment

We conducted a 1-year greenhouse experiment to assess the impact of nutrient manipulations on seedling growth, biomass partitioning, and leaf gas exchange between two fast growing Pinus taeda clones that differed in growth efficiency. After 1 year we observed significant treatment and treatment by clone effects on growth, biomass partitioning, and gas exchange parameters. Fertilization increased total seedling biomass 18% primarily through an increase in foliage and coarse-roots. Clones did not differ in total seedling biomass, however, clone 85 produced more stem than clone 93 leading to 37% greater stem:leaf, while clone 93 maintained more branch biomass. The logging residue treatment increased stem:leaf by 30%, but had no effect on total biomass or partitioning. Differences in leaf morphology resulted in significantly greater canopy leaf area in clone 93 than clone 85. Increased foliar N concentration from fertilization had only minor effects on specific photosynthesis under saturating light (A_Sat), but lowered stomatal conductance (g_s), transpiration (E), and internal to external CO₂ concentration ratio (C_i/C_a) as well as improved water use efficiency (WUE) independently of genotype. When gas exchange data was scaled to the canopy level both genotypes achieved similar canopy level CO₂ assimilation rates, but our data suggests they did this by different means. Although we did see a small effect of nutrient limitations in total canopy photosynthesis under saturating light (A_Canopy), A_Sat, and total leaf area (TLA), our foliar N concentration ([N]) indicated that our level of logging residue incorporation did not cause [N] to decrease below sufficiency limits. From a practical standpoint, a better understanding of strategies for capturing and partition C may lead to better selection of clonal material, thereby, optimizing productivity.     

The impact of extraction methodologies on the toxicity of sediments in the zebrafish (Danio rerio) embryo test

Purpose

Traditionally, methods for sediment extractions are characterised using chemical analyses. However, in order to evaluate sediment extracts with regard to biological effects and, thus, bioaccessibility, extraction methods have to be compared to effect data obtained from experiments with in situ exposure scenarios, i.e., sediment contact tests. This study compares four extraction methods and sediment contact test data from a previous project with respect to predictive power in the fish embryo test with zebrafish (Danio rerio).

Materials and methods

A natural and an artificial sediment spiked with a mixture of six organic pollutants (2,4-dinitrophenol, diuron, fluoranthene, nonylphenol, parathion and pentachlorophenol) were extracted using (a) membrane dialysis extraction (MDE), (b) a Soxhlet procedure, (c) hydroxypropyl-??-cyclodextrin (HPCD) or (d) Tenax^®-TA. Whereas the former two are regarded being exhaustive with respect to non-covalently bound contaminants, the latter two are considered to predict bioaccessibility. Resulting extracts were tested in the fish embryo assay with D. rerio for embryotoxic and teratogenic potential.

Results and discussion

Mortalities caused by organic extracts from Soxhlet extraction and MDE were high. However, HPCD extracts turned out to be at least as effective as extracts obtained with these two methods. One possible reason might be short ageing time of the spiked sediments. Only Tenax^®-TA extracts gave results comparable to the sediment contact assay for natural sediment, but revealed low reproducibility. Significant differences between natural and artificial sediment were found for extracts obtained with techniques using native (i.e., non-freeze-dried) sediments, i.e., HPCD and Tenax^®-TA. In contrast, MDE and Soxhlet extracts used freeze-dried sediment and did not differentiate between natural and artificial sediment. Therefore, freeze-drying has likely altered and equalised sediment properties that influence accessibility, such as composition of bacterial communities and organic matter quality.

Conclusions

Four extraction methods were successfully characterised with respect to their stringency and predictiveness for bioaccessibility. MDE was confirmed as an alternative to Soxhlet extraction. High mortalities induced by HPCD extracts underline the need to include ageing into consideration when assessing sediments. Although Tenax^®-TA may basically be used to predict bioaccessibility in the fish embryo test, the high variability observed warrants further investigation of the relation between effect and extractability. Apparently, freeze-drying can severely affect sediment properties, potentially eliminating individual properties of natural sediments.