Isolation and characterization of bone marrow-derived equine mesenchymal stem cells

OBJECTIVE: To isolate and characterize bone marrow-derived equine mesenchymal stem cells (MSCs) for possible future therapeutic applications in horses. SAMPLE POPULATION: Equine MSCs were isolated from bone marrow aspirates obtained from the sternum of 30 donor horses. PROCEDURES: Cells were cultured in medium (alpha-minimum essential medium) with a fetal calf serum content of 20%. Equine MSC features were analyzed to determine selfrenewing and differentiation capacity. For potential therapeutic applications, the migratory potential of equine MSCs was determined. An adenoviral vector was used to determine the transduction rate of equine MSCs. RESULTS: Equine MSCs can be culture-expanded. Equine MSCs undergo cryopreservation in liquid nitrogen without altering morphologic characteristics. Furthermore, equine MSCs maintain their ability to proliferate and differentiate after thawing. Immunocytochemically, the expression of the stem cell marker CD90 can be detected on equine MSCs. The multilineage differentiation potential of equine MSCs was revealed by their ability to undergo adipogenic, osteogenic, and chondrogenic differentiation. CONCLUSIONS AND CLINICAL RELEVANCE: Our data indicate that bone marrow-derived stromal cells of horses can be characterized as MSCs. Equine MSCs have a high transduction rate and migratory potential and adapt to scaffold material in culture. As an autologous cell population, equine MSCs can be regarded as a promising cell population for tissue engineering in lesions of the musculoskeletal system in horses.     

Serum chemistry alterations in Alaskan sled dogs during five successive days of prolonged endurance exercise

OBJECTIVE: To determine the impact of successive days of endurance exercise on select serum chemistry values in conditioned Alaskan sled dogs. DESIGN: Prospective cohort study. ANIMALS: 10 conditioned Alaskan sled dogs. PROCEDURES: All dogs ran 160 km/d for 5 consecutive days. Serum was obtained prior to exercise and immediately after each exercise run; all samples were obtained before dogs were fed. Serum electrolyte, mineral, protein, total bilirubin, urea nitrogen, creatinine, and cardiac troponin-I concentrations and serum alkaline phosphatase, alanine aminotransferase, creatine kinase, and aspartate aminotransferase activities were measured. Data were analyzed by means of analysis of covariance for a randomized complete block design with dog as a blocking variable, time as a covariate, and distance run as the treatment of interest. Least square mean values were compared with values obtained prior to exercise, and linear and quadratic contrasts were examined. RESULTS: Serum globulin concentration was low prior to exercise (mean +/- SD, 2.2 +/- 0.3g/dL) and progressively decreased as exercise continued. Exercise was associated with increases in serum chloride, urea nitrogen, and cardiac troponin-I concentrations and serum alanine aminotransferase, creatine kinase, and aspartate aminotransferase activities and with progressive decreases in serum potassium, total protein, and albumin concentrations. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that multiple successive days of endurance exercise resulted in mild aberrations in serum chemistry variables in conditioned sled dogs. Changes likely reflected the metabolic stresses of prolonged endurance exercise as well as dietary composition. Hypoglobulinemia in resting, conditioned sled dogs may reflect the immunosuppressive or catabolic effects of intense endurance training.     

Effect of high-pressure hot-water washing treatment on fruit quality, insects, and disease in apples and pears: Part I. System description and the effect on fruit quality of ‘d’Anjou’ pears

A manually operated high-pressure hot-water washing system consisting of a boiler, hot-water mixing tank, contact loop, heat exchanger, spray mixing tank, high-pressure hot-water washing manifold, low-pressure fresh water rinse manifold, and pressure pump was constructed and installed in a packingline. The system developed 20–50 °C washing water at pressures up to 980 kPa. ‘d’Anjou’ pears (Pyrus communis L.), shortly after harvest, and after storage for 3 and 4 months in regular air (RA) or for 4, 7 and 8 months in controlled atmosphere (CA) at −1 °C were washed through the packingline with different wetting agents (0.1% Silwet, 0.01 and 0.1% Defoamer, and water), water pressures (regular and high-pressure (210–980 kPa)), water temperatures (control (tap water, 4–22 °C), 40 °C, and 50 °C), and brushes (soft and firm), respectively. The effect of the washing conditions on fruit quality was investigated after 1 month of storage at −1 °C to simulate shipping condition, and then again after 1 week at 20 °C to simulate marketing condition. Hot-water caused severe heat scald. When nozzle temperature was 50 °C, the incidence of heat scald increased to over 50% for the fruit stored in RA for 3 months. Combined with hot-water, 540 kPa high-pressure washing increased the incidence of friction discoloration. There were lower incidences of friction discoloration and heat scald for fruit stored in CA for 7 months, in comparison to that in RA for 3 months. However, those fruit did not ripen properly as indicated by a high extractable juice content. Fruit washed at harvest had minor incidences of friction discoloration regardless of different brushes, water pressures, and wetting agents. Fruit washed after storages in either 4 months RA or 4 or 8 months CA suffered a high incidence of friction discoloration including scuffing symptoms and pressure marking. The firm brushes caused a higher incidence of friction discoloration mainly because of scuffing symptoms. However, no differences were found between different water pressures and wetting agents with respect to friction discoloration. Fruit stored for 4 months RA suffered 26–28% friction discoloration in comparison to 16–18% in CA stored fruit with firm brush washing. Extended CA storage increased friction discoloration even with soft brush washing. The results suggest that a washing system with high-pressure spray, <30 °C warm water, wetting agent Defoamer and rotating soft brushes were significantly effective in removing surface pests and decay control without causing internal or external damage of fruit.     

A cytokinin perception mutant colonized by Rhizobium in the absence of nodule organogenesis

In legumes, Nod-factor signaling by rhizobia initiates the development of the nitrogen-fixing nodule symbiosis, but the direct cell division stimulus that brings about nodule primordia inception in the root cortex remains obscure. We showed that Lotus japonicus plants homozygous for a mutation in the HYPERINFECTED 1 (HIT1) locus exhibit abundant infection-thread formation but fail to initiate timely cortical cell divisions in response to rhizobial signaling. We demonstrated that the corresponding gene encodes a cytokinin receptor that is required for the activation of the nodule inception regulator Nin and nodule organogenesis.     

Molecular phylogenetics of mastodon and Tyrannosaurus rex

We report a molecular phylogeny for a nonavian dinosaur, extending our knowledge of trait evolution within nonavian dinosaurs into the macromolecular level of biological organization. Fragments of collagen alpha1(I) and alpha2(I) proteins extracted from fossil bones of Tyrannosaurus rex and Mammut americanum (mastodon) were analyzed with a variety of phylogenetic methods. Despite missing sequence data, the mastodon groups with elephant and the T. rex groups with birds, consistent with predictions based on genetic and morphological data for mastodon and on morphological data for T. rex. Our findings suggest that molecular data from long-extinct organisms may have the potential for resolving relationships at critical areas in the vertebrate evolutionary tree that have, so far, been phylogenetically intractable.     

Inactivation of TNF signaling by rationally designed dominant-negative TNF variants

Tumor necrosis factor (TNF) is a key regulator of inflammatory responses and has been implicated in many pathological conditions. We used structure-based design to engineer variant TNF proteins that rapidly form heterotrimers with native TNF to give complexes that neither bind to nor stimulate signaling through TNF receptors. Thus, TNF is inactivated by sequestration. Dominant-negative TNFs represent a possible approach to anti-inflammatory biotherapeutics, and experiments in animal models show that the strategy can attenuate TNF-mediated pathology. Similar rational design could be used to engineer inhibitors of additional TNF superfamily cytokines as well as other multimeric ligands.     

Island biology and ecosystem functioning in epiphytic soil communities

Although island attributes such as size and accessibility to colonizing organisms can influence community structure, the consequences of these for ecosystem functioning are little understood. A study of the suspended soils of spatially discrete epiphytes or treetop "islands" in the canopies of New Zealand rainforest trees revealed that different components of the decomposer community responded either positively or negatively to island size, as well as to the tree species that the islands occurred in. This in turn led to important differences between islands in the rates of ecosystem processes driven by the decomposer biota. This system serves as a model for better understanding how attributes of both real and habitat islands may affect key ecosystem functions through determining the community structure of organisms that drive these functions.     

Plant mycoplasmas: a cultivable spiroplasma causes corn stunt disease

A spiroplasma can be isolated and grown continuously in cell-free medium from stunted corn or from Drosophila injected with sap expressed from diseased corn. The organism is serologically related to, but not identical with, Spiroplasma citri, the causative agent of citrus stubborn disease. Leafhopers injected with cultured organisms induced typical symptoms of the corn stunt disease when placed on previously healthy corn plants.