Development of AFLP and RAPD markers linked to a locus associated with twisted growth in corkscrew willow (Salix matsudana 'Tortuosa')

Salix matsudana Koidz. cultivar 'Tortuosa' (corkscrew willow) is characterized by extensive stem bending and curling of leaves. To investigate the genetic basis of this trait, controlled crosses were made between a corkscrew female (S. matsudana 'Tortuosa') and a straight-stemmed, wild-type male (Salix alba L. Clone 99010). Seventy-seven seedlings from this family (ID 99270) were grown in the field for phenotypic observation. Among the progeny, 39 had straight stems and leaves and 38 had bent stems and curled leaves, suggesting that a dominant allele at a single locus controls this phenotype. As a first step in characterizing the locus, we searched for amplified fragment length polymorphism (AFLP) and randomly amplified polymorphic DNA (RAPD) markers linked to the tortuosa allele using bulked segregant analysis. Samples of DNA from 10 corkscrew individuals were combined to produce a corkscrew pool, and DNA from 10 straight progeny was combined to make a wild-type pool. Sixty-four AFLP primer combinations and 640 RAPD primers were screened to identify marker bands amplified from the corkscrew parent and progeny pool, but not from the wild-type parent or progeny pool. An AFLP marker and a RAPD marker linked to and flanking the tortuosa locus were placed on a preliminary linkage map constructed based on segregation among the 77 progeny. Sectioning and analysis of shoot tips revealed that the corkscrew phenotype is associated with vascular cell collapse, smaller cell size in regions near the cambium and less developed phloem fibers than in wild-type progeny. Identification of a gene associated with this trait could lead to greater understanding of the control of normal stem development in woody plants.     

Mechanical measures of uncooked beef longissimus muscle can predict sensory panel tenderness and Warner-Bratzler shear force of cooked steaks

Two experiments were conducted to investigate mechanical measures of tenderness on uncooked USDA Select longissimus muscle as a means to predict Warner-Bratzler shear force (WBSF) and trained sensory panel tenderness (SPT) of cooked steaks. In Exp. 1, strip loins (n = 24) were aged 14 d postmortem and fabricated into steaks (2.54 cm). Medial, center, and lateral locations within uncooked steaks were evaluated by a plumb bob device and correlated with WBSF and SPT of cooked steaks. In Exp. 2, 24 strip loins were used to evaluate how well plumb bob and needle probe devices used on uncooked steaks predicted WBSF and SPT of cooked steaks. At 2 d postmortem, two steaks were fabricated from the anterior end. One uncooked steak (2.54 cm) was assigned to the plumb bob treatment and the other uncooked steak (5.08 cm) was assigned to needle probe treatment. At 14 d postmortem, one uncooked steak (5.08 cm) was assigned to needle probe treatment, a second uncooked steak (2.54 cm) was assigned to plumb bob treatment, whereas the remaining steaks (2.54 cm) were cooked and evaluated by a trained sensory panel and WBSF device. In Exp. 1, average plumb bob values were negatively correlated (P < 0.05) to average SPT scores (r = -0.48). However, correlations between WBSF and plumb bob values for medial, lateral, and average of all sections were not significant (P > 0.05). In Exp. 2, regression models to predict SPT from needle probe and plumb bob measurements individually taken at 2 d postmortem had R2 of 0.54 and 0.51, respectively. Combining needle probe and plumb bob measurements resulted in an R2 of 0.76; when quadratic terms for both variables were in the model, the R2 was 0.80. Regressing needle probe and plumb bob measurements at 2 d postmortem with WBSF produced R2 values of 0.51 and 0.45, respectively. If linear terms of both probes were combined to predict WBSF, the R2 increased to 0.77. An equation to predict WBSF, including both the linear and quadratic terms of needle probe and plumb bob measurements, resulted in an R2 of 0.84. Using plumb bob and needle probe devices on uncooked longissimus muscle at 2 d postmortem can predict cooked WBSF and SPT of USDA Select Grade steaks at 14 d postmortem.     

Negative-pressure pulmonary edema complicated by acute respiratory distress syndrome in an orangutan (Pongo pygmaeus abelii).

A 22-yr-old, 86-kg, morbidly obese female orangutan (Pongo pygmaeus abelii) was immobilized and transported to the Denver Zoological Gardens hospital for a routine physical examination. Immediately after arriving at the hospital, cyanosis and apparent inadequate ventilatory efforts were noted. Clinically significant hypoxia occurred despite attempts to ventilate the orangutan through face mask, and attempts to place an endotracheal tube began. A large volume of pink-tinged frothy fluid flowed from the trachea when the laryngoscope was inserted into the oropharynx. Severe pulmonary edema due to negative-pressure pulmonary edema, precipitating life-threatening hypoxia was suspected. The orangutan was maintained on a mechanical ventilator using the neuromuscular blocking agent cisatracurium besylate and sedation with periodic doses of isoflurane and midazolam for 48 hr. Positive end-expiratory pressure was used while the orangutan was ventilated mechanically to improve respiratory function. The edema and hypoxia improved, but respiratory arrest ensued 30 min after extubation, when the orangutan was removed from mechanical ventilation. Necropsy and histopathology demonstrated that serious lung injury had led to acute respiratory distress syndrome.     

Effects of cromolyn in horses with chronic obstructive pulmonary disease

The effect of disodium cromoglycate (cromolyn) in preventing the pulmonary dysfunction caused by the inhalation of barn and hay dust was studied in 5 horses with confirmed chronic obstructive pulmonary disease (COPD). The horses were studied before (Con) and after exposure to hay and dust allergens (Expos) and after pretreatment with cromolyn followed by exposure (Cr-Expos).There was a significant reduction in PaO₂ from 86.8±8.3 to 73.1±8.8 when the horses were exposed to hay and dust allergens. The PaO₂ after pretreatment with cromolyn and exposures was 78.1±5.5. There were no significant changes in PaCO₂, FRC, pH and A-aDO₂ when the Con, Expos, and Cr-Expos periods were compared.There were significant increases in V_E from a control value of 77.9±18.2 to 128.7±55.1 and 133.7±17.1 L/min during the exposures, which was due primarily to increases in respiratory frequency. Respiratory dead space (V_D/V_T) increased from 0.55±0.10 to 0.71±0.07 and 0.65±0.05, and alveolar ventilation (V_E) remained constant. Pulmonary resistance (R_L) and transpulmonary pressure (P_tr) increased from a control of 0.77±0.28 cm H₂O/L/sec and 7.73±3.38 cm H₂O to 2.93±1.01 and 20.17±4.81 during the Expos period and tidal volume (V_T) fell from 7.5±1.0 to 5.7±1.3 L. The pre-treatment with cromolyn before exposures significantly reduced the increase in R_L and P_tr and returned V_T to Con levels.Supported by Fisons, Ltd, Pharmaceutical Division and the Pennsylvania State Horse Racing Commission     

Dietary fibre in the diet of the growing pig: effects on apparent digestibility and resultant implications for its use in reducing the incidence of oesophagogastric parakeratosis.

The effects on apparent digestibility of increasing dietary fibre concentrations in barley-based diets by substituting for barley either bran, oatmeal by-product, guar gum or pectin were studied using 72 pigs. Increasing substitutions of bran up to a maximum of 300 g kg-1 gave progressive decreases in apparent digestibility of dry matter, ether extract and gross energy fractions and in digestible and metabolisable energy contents, progressive increases in apparent digestibility of modified acid detergent fibre but had no consistent effect on either nitrogen or extract free of nitrogen fractions. Increasing substitution of oatmeal by-product up to the same maximum gave significant linear decreases in apparent digestibility of dry matter, extract free of nitrogen and gross energy, significant depressions in apparent digestibility of modified acid detergent fibre and significant increases in ether extract. Guar gum and pectin, both substituted to a maximum of 50 g kg-1 had no significant effects on apparent digestibility. The results are discussed in the context of the concentrations of dietary fibre needed to reduce the incidence of oesophagogastric parakeratosis.     

The Effect of Temperature on Production of Gonads by the Sea Urchin Strongylocentrotus franciscanus Fed Natural and Prepared Diets

Strongylocentrotus franciscanus (248 ± 76 g wet body weight, mean and standard deviation) were maintained in the laboratory at 12.9 and 16.1 C and fed either Nereocystis luetkeana or a prepared feed for 4 mo (July through October). The size and histological condition of the gonads of an initial sample, the experimental animals, and a sample from the commercial fishery were evaluated. The gonad index of experimental S. franciscanus increased from an initial 3.4%± 1.4 to 19.2%± 3.3 (mean and standard deviation) for the four treatments. The feeding rate was greater for individuals fed N. luetkeana and for both foods at the higher temperature. Gonad production was not significantly different among the experimental treatments, indicating the prepared feed was nutritionally superior to the algal diet and an increase in catabolism at the higher temperature balanced the increase in food intake. Histological preparations determined the urchin gonads developed from an initial spent condition to the premature stage for both experimental and fishery samples, indicating neither food or temperature affected gametogenesis.     

Randomized, controlled study of inhaled fluticasone propionate, oral administration of prednisone, and environmental management of horses with recurrent airway obstruction

OBJECTIVE: To determine whether administration of glucocorticoids provides additional benefits to environmental management of horses with recurrent airway obstruction (RAO). ANIMALS: 28 horses with RAO. PROCEDURE: Horses were classified as having mild, moderate, or severe RAO. Within each category, horses were randomly assigned to receive inhaled fluticasone propionate, inhaled control substance, or oral administration of prednisone. During the 4-week study, horses were maintained outdoors and fed a pelleted feed. Clinical scores, pulmonary function, results of cytologic examination of bronchoalveolar lavage fluid (BALF), and adrenal gland function were determined before and 2 and 4 weeks after initiation of treatment. RESULTS: Clinical score and pulmonary function of all RAO-affected horses improved during the treatment period. After 4 weeks, clinical scores and pulmonary function of horses treated with a glucocorticoid were not different from those for the control treatment. In horses with severe RAO, treatment with fluticasone for 2 weeks resulted in significantly greater improvement in pulmonary function, compared with pulmonary function after treatment with prednisone or the control substance. Treatment with a glucocorticoid for 4 weeks and a low-dust environment did not have any effect on cellular content of BALF Treatment with prednisone for 2 weeks resulted in a significant decrease in serum cortisol concentration, compared with concentrations after administration of fluticasone or the control substance. CONCLUSIONS AND CLINICAL RELEVANCE: Environmental management is the most important factor in the treatment of horses with RAO. Early treatment with inhaled fluticasone can help accelerate recovery of horses with severe RAO.     

In vivo expression of GFP transgene delivered via a replicating feline leukemia virus

We previously described replication-competent feline leukemia virus (FeLV) vectors with high-level and stable expression of a green fluorescent protein (GFP) or a suicide transgene in cell cultures in vitro. Considering that FeLV might potentially be used to deliver therapeutic genes in vivo, we first evaluated the expression of the GFP gene introduced in cats by the FeLV, Rickard subgroup A (FRA) construct. Eight newborn kittens were either inoculated with pFRA-GFP plasmid DNA intradermally, or challenged intraperitoneally with FRA-GFP-infected feline fibroblasts. During a 12-week observation period, five cats were shown to be progressively viremic. Quantitative PCR and RT-PCR analyses of plasma and tissue samples from these cats showed that GFP was retained in FeLV DNA or RNA to a variable degree, ranging from 0.002 to 27.890%. Tissue DNA samples were analyzed by PCR for the status of GFP and the env-transgene complex. While the proviruses carrying the GFP transgene were shown to be minor species, all tissues, however, retained the full-length GFP transgene. Despite the occurrence of predominant species with various deletions in the viral genome, approximately 1–3% of the total cell population was GFP-positive in the lymphoid tissues as visualized by laser confocal microscopy. Co-localization of immunofluorescent cells indicated that CD3-positive T cells, dendritic cells and macrophages were the major targets for GFP expression. These findings on the detectable in vivo expression of GFP for as long as a period of 3 months could be viewed positively for contemplating a therapeutic strategy for control of FeLV infection in the cats.