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.     

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.     

May the Force (Field) Be with You: On the Importance of Conformational Searches in the Prediction of NMR Chemical Shifts

NMR data prediction is increasingly important in structure elucidation. The impact of force field selection was assessed, along with geometry and energy cutoffs. Based on the conclusions, we propose a new approach named mix-J-DP4, which provides a remarkable increase in the confidence level of complex stereochemical assignments—100% in our molecular test set—with a very modest increment in computational cost.     

Applying Local Measures of Spatial Heterogeneity to Landsat-TM Images for Predicting Wildfire Occurrence in Mediterranean Landscapes

In mountainous Mediterranean regions, land abandonment processes in past decades are hypothesized to trigger secondary vegetal succession and homogenization, which in recent years has increased the size of burned areas. We conducted an analysis of temporal changes in landscape vegetal spatial pattern over a 15-year period (1984–1998) in a rural area of 672.3 km² in Eastern Spain to investigate the relationship between local landscape heterogeneity and wildfire occurrence. Heterogeneity was analyzed from textural metrics derived from non-classified remote sensing data at several periods, and was related to wildfire history in the study area. Several neural network models found significant relationships between local spatial pattern and future fire occurrence. In this study, sensitivity analysis of the texture variables suggested that fire occurrence, estimated as probability of burning in the near future, increased where local homogeneity was higher.     

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.     

Diversity in White Lupin (<Emphasis Type="Italic">Lupinus albus</Emphasis> L.) Landraces from Northwest Iberianplateau

All the accessions (35) of white lupin (Lupinus albus L.) landraces collected from northwest Iberian plateau and maintained at the Spanish germplasm collection (CRF of INIA), were studied with the general objective of assessing the biodiversity of these landraces and to ascertain their value as genetic resources for the development of germplasm adapted to the areas where they were collected with long and chilly winters. The characterization study comprised 51 qualitative and 50 quantitative characters. Quantitative parameters were analyzed by Principal Components Analysis (PCA). The 2-dimensional plot (49.3% of cumulative variability) formed a main group of accessions and 4 outliers (#1, #9, #27 and #28) separated in the first PC. The characters with a higher contribution to the first PC were inflorescence length, leaf petiole length and leaf central foliole area. The presence of alkaloids, the percentage of plants killed by soil borne fungal diseases, the phenology, the yield per plant and yield components were also assessed. Twenty six accessions (the main group in the PCA plot) showed high homogeneity, with the following highlights: bitterness, indeterminate growth habit, early ripeness and spring sowing with the exception of #18, in which seeding date is unknown and it had a later phenology. By contrast accessions #9 and #27 were bitter of autumn sowing with determinate growing habit and they had the longest values of primary inflorescence length and a later phenology when they were sown in spring. Average values for yield and yield components showed a broad range of variation between accessions. The main yield component was the number of pods per plant. Accession #22 had the highest mean seed mass, although this had not significant influence in yield. Accession #17 showed the highest yield and #1 the lowest. This last also had the lowest values of inflorescence length, leaf petiole length and leaf central foliole area. No significant differences of resistance to soil-borne fungi were found between accessions. The year had a significant effect in the phenology, yield and yield components except for number of seeds per pod. The studied material might be of interest for the development of spring sowing germplasm adapted to north Iberian peninsula.     

Usefulness of amino acid composition to discriminate between honeydew and floral honeys. Application to honeys from a small geographic area

With the aim of finding methods that could constitute a solid alternative to melissopalynological and physicochemical analyses to determine the botanical origin (floral or honeydew) of honeys, the free amino acid content of 46 honey samples has been determined. The honeys were collected in a small geographic area of approximately 2000 km(2) in central Spain. Twenty-seven honey samples were classified as floral and 19 as honeydew according to their palynological and physicochemical analyses. The resulting data have been subjected to different multivariant analysis techniques. One hundred percent of honey samples have been correctly classified into either the floral or the honeydew groups, according to their content in glutamic acid and tryptophan. It is concluded that free amino acids are good indicators of the botanical origin of honeys, saving time compared with more tedious analyses.     

Antiradical activity of water soluble components in common diet vegetables

The antiradical activity of water-soluble components contained in mushrooms (Psalliota campestris), onions (Allium cepa), white cabbage (Brassica oleracea var. alba), and yellow bell peppers (Capsicum annuum) against hydroxyl radicals was tested in a chemical and biological system. The vegetable juices were obtained by centrifugation of a vegetable homogenate processed at 2 degrees C or heated at 102 degrees C. The chemical system consisted of a buffered reaction mixture composed of Fe(III)-EDTA, 2-deoxy-D-ribose, ascorbic acid, and H(2)O(2) generating the hydroxyl radical. The antiradical activity was expressed as an inhibition of deoxyribose degradation. The biological system consisted of IMR32 neuroblastoma cells exposed to H(2)O(2) in the presence or absence of the vegetable juices. Cells were pretreated for either 24 h or 1 h with the vegetable juices, and reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) was used as a cell viability assay. All vegetable juices inhibited the degradation of deoxyribose and increased the viability of H(2)O(2) treated cells. Raw mushroom juice proved to be the most active in both cases. Boiling significantly affected the activity of mushroom juice, but did not change significantly the effect on onions and yellow bell peppers, and partially increased the activity of white cabbage juice. Mushroom antiradical activity was also confirmed by a cytofluorimetric analysis.     

A new large‐scale and cost‐effective restoration method for cold‐water coral gardens

1. Gorgonians dwelling on the Mediterranean continental shelf are among the most frequent fishing bycatch taxa. These species display several traits, such as long lifespans and slow growth, which make them very vulnerable to the impacts caused by fishing activities with far‐reaching and long‐lasting effects.
2. Hence, restoration and mitigation actions are crucial to enhance and speed up the natural recovery of damaged cold‐water coral gardens. Given the growing concern to develop effective and affordable restoration actions, the present study aims to propose and technically validate a new large‐scale and cost‐effective restoration method.
3. This technique, named ‘badminton method', consists of attaching bycatch Eunicella cavolini colonies to cobble supports and returning them to the continental shelf by gently throwing the gorgonian transplants directly from a boat.
4. Two consecutive field experiments were conducted in order to find the best cobble type support and gorgonian size to be used: first, to evaluate the landing efficiency of gorgonian transplants at different depths (from 5 to 30 m) and second, to evaluate their capability to maintain a correct position over time.
5. Natural cobbles with large gorgonians attached were the best option. Field results and modelling approaches suggest that the transplants would correctly land on the continental shelf seabed in a predicted area of around 60 m². Moreover, they would successfully maintain an upright position ensuring gorgonian survival over time.
6. The success of this method highlights the feasibility of large‐scale and low‐cost restoration actions with promising results for the conservation and recovery of cold‐water coral gardens.