Effects of recombinant canine granulocyte colony-stimulating factor on white blood cell production in clinically normal and neutropenic dogs.

Recombinant canine granulocyte colony-stimulating factor (rcG-CSF) was administered to clinically normal dogs, cyclic-hematopoietic dogs, and dogs undergoing autologous bone marrow transplantation, to determine whether rcG-CSF could be used to stimulate WBC production and function in normal and neutropenic dogs. To the normal dogs, rcG-CSF was administered by SC injection at rates of 1 microgram/kg of body weight, q 12 h; 2 micrograms/kg, q 12 h; or 5 micrograms/kg, q 12 h. A significant dose-dependent increase in the WBC count resulted from the stimulation of bone marrow progenitor cells. The increased WBC count was characterized by mature neutrophilia and monocytosis. Neutrophil myeloperoxidase and phagocytic activity were normal in rcG-CSF-treated normal dogs, demonstrating the production of normal functional neutrophils in response to rcG-CSF treatment. Recombinant canine G-CSF prevented neutropenia and associated clinical signs but did not completely eliminate the cycling of neutrophils in cyclic-hematopoietic dogs when it was administered at rates of 1 microgram/kg, q 12 h, and 2.5 micrograms/kg, q 12 h. The time to bone marrow reconstitution was not decreased in dogs treated with rcG-CSF at a rate of 2.5 micrograms/kg, q 12 h, for 13 days following autologous bone marrow transplantation. On the basis of our findings, we suggest that treatment with rcG-CSF is an effective way to stimulate myelopoiesis in dogs, but that the dose of rcG-CSF required to stimulate WBC production will vary depending on the cause of neutropenia. Recombinant canine G-CSF should be useful in stimulating production and maintaining function of WBC for treatment of clinical diseases seen commonly in veterinary practice.     

The effect of dose and concentration on D-xylose absorption in healthy,immature dogs

Nine combinations of dosages and concentrations of D-xylose were given orally to eight clinically normal, immature dogs. The concentrations and dosages of D-xylose consisted of 5%, 10%, and 20% at 250 mg/kg, 500 mg/kg, and 750 mg/kg. Serum samples were collected at 0, 30, 60, 90, 120, and 180 minutes. Serum xylose was quantitated using the phloroglucinol microassay technique. A peak in serum xylose concentration was seen for each treatment combination at 60 or 90 minutes after dosing. The dosage effect was important in influencing serum xylose values (P < 0.0001). As the test solution dosages increased from 250 mg/kg to 500 mg/kg and 750 mg/kg, serum xylose values (when dosage was analyzed over the length of the entire test) rose linearly (R(2) = 0.98). The treatment combinations of 5% and 20% xylose solutions dosed at 750 mg/kg produced the highest serum xylose values at the 60- and 90-minute peak intervals. The independent effect of concentration was significant (p < 0.001) but was overridden by the stronger dosage effect. Serum xylose concentrations varied little statistically (p > 0.05) when the 5%, 10%, and 20% solutions were compared at a specific dosage.     

Relationship between energy and protein intakes and laying characteristics in individually‐caged broiler breeder hens

1. Individually‐caged broiler hens, which had been reared on an advised rationing programme, were fed allowances of 1.88, 1.61, 1.32 or 1.13 MJ apparent metabolisable energy/bird d at four different protein intakes (27, 23, 19.5 or 16.5 g crude protein per bird d) from 21 to 60 weeks of age.

2. Age at first egg, body‐weight gain and egg production were affected by energy allowance. Birds on the lower energy allowances came into lay later than birds on the higher energy allowances and at a lower body weight.

3. Body‐weight gain decreased with decreasing energy allowance. The decrease in egg output in response to decreasing energy allowance resulted from more birds ceasing to lay and fewer birds laying on more than 3 d per week. Similar changes in the distribution of rates of lay were observed on each treatment as the flock aged.

4. The relationship between body‐weight gain and egg number on each treatment was negative from 21 to 36 weeks, but became less consistent with age.

5. Protein intake had little effect on body weight. At the lowest energy allowance, egg number and egg weight decreased with increasing protein allowance. This effect was not observed on the higher energy allowances.     

Effects of energy and protein allowances during lay on the reproductive performance of broiler breeder hens

1. Broiler parent stock were fed daily allowances of 1.88, 1.73 or 1.52 MJ apparent metabolisable energy (AME) per bird at two different daily protein intakes (27, 21.3 g crude protein (CP) per bird) or daily protein intakes of 24.6 and 19.4 g CP per bird at a daily energy intake of 1.88 MJ AME per bird from 21 to 64 weeks of age.

2. Body‐weight gain and carcass fat and water content increased and fertility decreased with increasing energy allowance. Maximum egg production occurred at an energy intake of 1.73 MJ AME/bird d.

3. Differences in egg weight and hatchability were related to differences in both energy and protein intake. The highest egg weight occurred at the highest allowance of energy and protein. Hatchability was depressed where the daily allowances of protein and energy were in a ratio of more than 15 g protein: 1 MJ AME.

4. Apart from egg size no significant effects on reproductive performance were observed when dietary protein intake was varied from 27 to 19.5 g/bird d.

5. Requirements of broiler breeder hens for protein during lay may be lower than previously thought. For the strain used a protein intake of 19.5 g/bird d appeared adequate provided essential amino acid concentrations were maintained.

6. The close relationships between body weight and energy allowance and the latter and egg production make body‐weight gain a useful guide to management. A body‐weight gain of about 1.1 kg from 21 to 36 weeks of age was associated with optimum performance in this study.     

Resistance to Leveillula taurica in the genus Capsicum

One hundred and sixty-two Capsicum genotypes were evaluated for powdery mildew (Leveillula taurica) resistance, following inoculations with a suspension of 5 × 10⁴ conidia mL⁻¹ on 10-leaved to 12-leaved plants. Genotypes were graded into five resistance classes, based on the areas under the disease progress curves calculated from disease incidence (percentage infected leaves per plant) and severity (total number of colonies per plant). Results revealed a continuum from resistance to susceptibility, with the majority (70%) of C. annuum materials being classified as moderately to highly susceptible to L. taurica. Conversely, C. baccatum, C. chinense and C. frutescens were most often resistant, indicating that resistance to L. taurica among Capsicum species is found mainly outside the C. annuum taxon. Nevertheless, some resistant C. annuum material was identified that may be useful for resistance breeding. Eight genotypes were identified as immune to the pathogen: H-V-12 and 4638 (previously reported), and CNPH 36, 38, 50, 52, 279 and 288. Only H-V-12 and 4638 are C. annuum, while all others belong to the C. baccatum taxon. Latent period of disease on a set of commercial sweet pepper genotypes varied, indicating diverse levels of polygenic resistance. The latent period progressively reduced with plant maturity, from 14·3 days in plants at the mid-vegetative stage to 8·6 days in plants at the fruiting stage. Young plants of all commercial genotypes tested at the early vegetative stage were immune, irrespective of the reaction of the genotype at later stages, demonstrating widespread juvenile resistance to L. taurica in the Capsicum germplasm. Inoculation of plants of different botanical taxa with a local isolate indicated a wide host range. Some hosts, including tomato (Lycopersicon esculentum), artichoke (Cynara scolymus) and poinsettia (Euphorbia pulcherrima), produced large amounts of secondary inoculum. Other hosts included okra (Abelmoschus esculentus), eggplant (Solanum melongena), cucumber (Cucumis sativus), Solanum gilo, Chenopodium ambrosioides and Nicandra physaloides.     

Monitoring response to aminobisphosphonate therapy in canine osteosarcoma using dual energy x‐ray absorptiometry (dexa)

Introduction: Palliative therapy is essential to improve the quality of life of dogs with osteosarcoma (OSA), when definitive therapy is not considered a valid option. Bisphosphonates, a novel class of antiosteoclastic drugs, are widely used in humans for several painful osteolytic conditions. Dual energy x‐ray absorptiometry (DEXA) is recognized as a reliable tool to measure bone mineral density (BMD), and to monitor treatment response to bisphosphonates in humans. A prospective evaluation of pamidronate, an injectable aminobisphosphonate, is ongoing in dogs with appendicular OSA. The potential value of DEXA for objective evaluation of BMD variations with palliative therapies is concurrently being assessed. Materials and Methods: Dogs with naturally occurring appendicular OSA treated with pamidronate constitute the patient population. A DEXA scan (QDR‐4500 W, Hologic, Bedford, MA) is performed on day 0 (baseline) and on every treatment day with pamidronate thereafter (every 28 days). For each dog, a whole body scan is performed, followed by a scan of the tumor, and contralateral normal bone. Three regions of interest are subsequently analyzed for BMD changes in tumor and normal bone. Statistical analysis was performed using Student t‐test and paired t‐tests, with significance being set at p < 0.05. Results: Nineteen dogs have been enrolled to date. Seven responders and 6 non‐responders have suitable data for analysis. A significant difference is observed (p = 0.04) between tumor BMD variations of responders and non‐responders at day 28 (mean variations +18.0% and ?4.6% respectively). The changes at day 28 are significant only in the responders (p = 0.038 vs p = 0.05 in non‐responders). When BMD of tumor and normal bone at day 84 is compared to day 0 in six responders, only tumor had a significant increase (p = 0.017 vs p = 0.279, respectively). Conclusions: Objective measurements of response to therapy are essential in pain palliation studies. Increased tumor bone BMD, as obtained by DEXA analysis, may correlate with subjective clinical improvement in pamidronate‐treated dogs with appendicular OSA.     

Frequent mutation of BAP1 in metastasizing uveal melanomas

Metastasis is a defining feature of malignant tumors and is the most common cause of cancer-related death, yet the genetics of metastasis are poorly understood. We used exome capture coupled with massively parallel sequencing to search for metastasis-related mutations in highly metastatic uveal melanomas of the eye. Inactivating somatic mutations were identified in the gene encoding BRCA1-associated protein 1 (BAP1) on chromosome 3p21.1 in 26 of 31 (84%) metastasizing tumors, including 15 mutations causing premature protein termination and 5 affecting its ubiquitin carboxyl-terminal hydrolase domain. One tumor harbored a frameshift mutation that was germline in origin, thus representing a susceptibility allele. These findings implicate loss of BAP1 in uveal melanoma metastasis and suggest that the BAP1 pathway may be a valuable therapeutic target.     

Recent Advances in the Synthesis of Marine-Derived Alkaloids via Enzymatic Reactions

Alkaloids are a large and structurally diverse group of marine-derived natural products. Most marine-derived alkaloids are biologically active and show promising applications in modern (agro)chemical, pharmaceutical, and fine chemical industries. Different approaches have been established to access these marine-derived alkaloids. Among these employed methods, biotechnological approaches, namely, (chemo)enzymatic synthesis, have significant potential for playing a central role in alkaloid production on an industrial scale. In this review, we discuss research progress on marine-derived alkaloid synthesis via enzymatic reactions and note the advantages and disadvantages of their applications for industrial production, as well as green chemistry for marine natural product research.