Isolation and characterization of pediatric canine bone marrow CD34+ cells

Historically, the dog has been a valuable model for bone marrow transplantation studies, with many of the advances achieved in the dog being directly transferable to human clinical bone marrow transplantation protocols. In addition, dogs are also a source of many well-characterized homologues of human genetic diseases, making them an ideal large animal model in which to evaluate gene therapy protocols. It is generally accepted that progenitor cells for many human hematopoietic cell lineages reside in the CD34+ fraction of cells from bone marrow, cord blood, or peripheral blood. In addition, CD34+ cells are the current targets for human gene therapy of diseases involving the hematopoietic system. In this study, we have isolated and characterized highly enriched populations of canine CD34+ cells isolated from dogs 1 week to 3 months of age. Bone marrow isolated from 2- to 3-week-old dogs contained up to 18% CD34+ cells and this high percentage dropped sharply with age. In in vitro 6-day liquid suspension cultures, CD34+ cells harvested from 3-week-old dogs expanded almost two times more than those from 3-month-old dogs and the cells from younger dogs were also more responsive to human Flt-3 ligand (Flt3L). In culture, the percent and number of CD34+ cells from both ages of dogs dropped sharply between 2 and 4 days, although the number of CD34+ cells at day 6 of culture was higher for cells harvested from the younger dogs. CD34+ cells harvested from both ages of dogs had similar enrichment and depletion values in CFU-GM methylcellulose assays. Canine CD34+/Rho123lo cells expressed c-kit mRNA while the CD34+/Rhohi cells did not. When transplanted to a sub-lethally irradiated recipient, CD34+ cells from 1- to 3-week-old dogs gave rise to both myeloid and lymphoid lineages in the periphery. This study demonstrates that canine CD34+ bone marrow cells have similar in vitro and in vivo characteristics as human CD34+ cells. In addition, ontogeny-related functional differences reported for human CD34+ cells appear to exist in the dog as well, suggesting pediatric CD34+ cells may be better targets for gene transfer than adult bone marrow. The demonstration of similarities between canine and human CD34+ cells enhances the dog as a large, preclinical model to evaluate strategies for improving bone marrow transplantation protocols, for gene therapy protocols that target CD34+ cells, and to study the engraftment potential of various cell populations that may contain hematopoietic progenitor cell activity.     

Human recombinant interferonalpha-2b for management of idiopathic recurrent superficial pyoderma in dogs: a pilot study

The oral use of human recombinant interferonalpha-2b at 1,000 IU/ml/day appeared to provide only a transient benefit as compared with placebo for management of idiopathic recurrent superficial pyoderma in dogs. Further investigation using a larger population of dogs is needed to determine whether interferon is effective for long-term treatment of this condition.     

Experimental evaluation of ophthalmic devices and solutions using rabbit models

OBJECTIVE: To analyze and compare the geometry of the anterior segment of rabbit and human eyes, with relevance for the evaluation of intraocular lenses, and to review rabbit models used in our laboratory for the evaluation of different ophthalmic devices and solutions. PROCEDURES: Fifteen rabbit and 15 human eyes (10 phakic and 5 pseudophakic/group) obtained postmortem were used. Anterior-posterior length, equatorial diameter, and white-to-white (corneal diameter) were measured with calipers. The eyes were then analyzed with a very high-frequency ultrasound (Artemis, Ultralink) for measurements of the anterior chamber depth, and anterior chamber and ciliary sulcus diameters. The capsular bag diameter was measured with calipers from a posterior view, and the diameter and thickness of the crystalline lenses were measured after their excision from the phakic eyes. RESULTS: Although the size of the rabbit eye is overall smaller than the size of the human eye, the dimensions of the anterior segment of rabbit eyes are generally larger. The differences between rabbit and human eyes were statistically significant (Wilcoxon rank sum test) in terms of anterior-posterior length, equatorial diameter, white-to-white measurements (P < 0.0001), anterior chamber diameter (P = 0.0004), ciliary sulcus diameter (P = 0.0012), and crystalline lens diameter and thickness (P = 0.0003). CONCLUSIONS: Experimental evaluation of design features of new phakic intraocular lenses in rabbit eyes may be inconclusive without adaptation of their size/design, contrary to the evaluation of new pseudophakic lenses by implantation in the capsular bag. The rabbit is a very valuable model for the experimental evaluation of different ophthalmic devices and solutions.     

Population genomics of early events in the ecological differentiation of bacteria

Genetic exchange is common among bacteria, but its effect on population diversity during ecological differentiation remains controversial. A fundamental question is whether advantageous mutations lead to selection of clonal genomes or, as in sexual eukaryotes, sweep through populations on their own. Here, we show that in two recently diverged populations of ocean bacteria, ecological differentiation has occurred akin to a sexual mechanism: A few genome regions have swept through subpopulations in a habitat-specific manner, accompanied by gradual separation of gene pools as evidenced by increased habitat specificity of the most recent recombinations. These findings reconcile previous, seemingly contradictory empirical observations of the genetic structure of bacterial populations and point to a more unified process of differentiation in bacteria and sexual eukaryotes than previously thought.     

Comparison of radiographic osteoarthritis scores in dogs less than 24 months or greater than 24 months following tibial plateau leveling osteotomy

Radiographic osteoarthritis scores were determined in 60 dogs up to 3 years following tibial plateau leveling osteotomy. Radiographs taken immediately following surgery and at long-term follow-up 1 to 3 years later were evaluated by 2 certified radiologists using a modified 32-point osteoarthritis scale. Changes in osteoarthritis scores were evaluated by paired t-tests and regression analysis. Sub-groups of dogs were formed to evaluate if osteoarthritis scores changed differently for follow-up periods of < 24 months compared with those > 24 months. There was a significant increase in osteoarthritis score from post-operative to follow-up evaluations for all dogs studied. Regression analysis of the relationship of months after surgery to change in osteoarthritis score was not significant. These results indicate that osteoarthritis did progress following tibial plateau leveling osteotomy, but not in a linear fashion over time.