The second-generation curcumin analogue RL71 elicits G2/M cell cycle arrest and apoptosis in canine osteosarcoma cells

Canine osteosarcoma is an aggressive cancer, comprising 85% of canine bone neoplasms. Current treatment practices of surgery and chemotherapy increase 1-year survival by only 45%. The curcumin analogue RL71, has demonstrated potent in vitro and in vivo efficacy in several models of human breast cancer through increased apoptosis and cell cycle arrest. Thus, the present study aimed to investigate efficacy of curcumin analogues in two canine osteosarcoma cell lines. Osteosarcoma cell viability was assessed using the sulforhodamine B assay and mechanisms of action were determined by analysing the levels of cell cycle and apoptotic regulatory proteins via Western blotting. Further evidence was obtained using flow cytometry to detect cell cycle distribution and the number of apoptotic cells. RL71 was the most potent curcumin analogue with EC₅₀ values of 0.64 ± 0.04 and 0.38 ± 0.009 μM (n = 3) in D-17 (commercial) and Gracie canine osteosarcoma cells, respectively. RL71 significantly increased the ratio of cleaved-caspase 3 to pro-caspase 3 and the level of apoptotic cells at the 2× and 5× EC₅₀ concentration (p < 0.001, n = 3). Furthermore, at the same concentration, RL71 significantly increased the number of cells in the G2/M phase. In conclusion, RL71 has potent cytotoxic activity in canine osteosarcoma cells triggering G2/M arrest and apoptosis at concentrations achievable in vivo. Future research should further investigate molecular mechanisms for these changes in other canine osteosarcoma cell lines prior to in vivo investigation.     

A comparative description of two farmer-managed irrigation systems in Nepal

The organization of two farmer-managed irrigation systems in the western hills of Nepal is described by examining the ways in which the activities of water allocation, water distribution, maintenance, and resource mobilization are performed. Due to the topography and environment, these two organizations are structured primarily to mobilize the large amount of labor required for maintenance of the intake and canal. Both organizations precisely define each member's water allocation. In one system, water is allocated in proportion to the area of an individual's land holding, while in the other water allocation is by purchased shares. These two cases were used to analyze the importance of the principle of water allocation for expansion of area irrigated and equity of access to irrigation. Evidence from the two systems shows that in this hill environment water allocation by purchased shares provides the individual incentive and an organizational mechanism for efficient development of irrigation resources. Expansion of area irrigated and equity of access to irrigation were found to be greater in the system which allocates water by purchased shares than where water was allocated in proportion to area irrigated.     

Potential farmer contributions to the design process: Indications from Indonesia

The paper argues that conventional irrigation design processes rely heavily on predetermined design criteria, often without allowing for interactions with farmers and revising of criteria during implementation. Six cases from a tertiary development project in North Sulawesi, Indonesia are described, where farmers altered what had been designed and constructed by contracting and agency engineers. Through field inspections and farmer interviews, the farmers' alternative set of design criteria was elicited to explain why the farmers changed what had been constructed. A total of 27 cases are analyzed to show the relative importance of:

Serologic survey of domestic felids in the Petén region of Guatemala.

Blood samples were analyzed from 30 domestic cats (Felis domesticus) from the Petén region of Guatemala to determine the seroprevalence of common pathogens that may pose a potential risk to native wild felids. Eight of the cats had been vaccinated previously; however, owners were unable to fully describe the type of vaccine and date of administration. In addition, blood samples were obtained from two captive margays (Leopardus wiedii). Samples were tested for antibodies to feline immunodeficiency virus, Dirofilaria immitis, feline panleukopenia virus, feline herpesvirus, feline coronavirus, canine distemper virus, and Toxoplasma gondii and for feline leukemia virus (FeLV) antigen. Fifty percent or more of the cats sampled were seropositive for feline herpesvirus (22 of 30), feline panleukopenia (15 of 30), and T. gondii (16 of 30). Five cats were positive for FeLV antigen. Both margays were seropositive for feline coronavirus and one was strongly seropositive to T. gondii. All animals were seronegative for D. immitis. This survey provides preliminary information about feline diseases endemic to the Petén region.     

Myelodysplastic syndrome with sideroblastic differentiation in a dog

A dog with myelodysplastic syndrome had microcytic hypochromic anemia, with siderocytes, poikilocytosis, hypersegmented neutrophils and giant platelets in peripheral blood. In bone marrow, the erythroid series showed immaturity, asynchronous maturation and sideroblasts. Dysgranulopoiesis and dysthrombopoiesis also were present, and hemosiderin was increased. Serum iron concentration was high, and both iron deficiency and lead toxicity were excluded as the cause of dyscrasia. This case represents a unique variant of myelodysplastic syndrome, best described as sideroblastic myelodysplasia. We propose the terms dyserythropoiesis, sideroblastic dyserythropoiesis, dyserythropoiesis with excess blasts, myelodysplasia and sideroblastic myelodysplasia to describe and categorize myelodysplastic syndromes in dogs.     

Forest fragmentation in Loudoun County,Virginia, USA evaluated with multitemporal Landsat imagery

In order to study forest fragmentation in the Virginia, USA Piedmont, a series of Landsat images from 1973, 1987, and 1999 covering a rapidly developing area (Loudoun County) was used to classify forest from non-forest. The classified images were analyzed using a geographic information system (GIS) to determine the spatial and temporal patterns of fragmentation, and to relate these patterns to infrared radiance provided by Landsat Enhanced Thematic Mapper Plus (ETM+) band 6. The analysis was concentrated on eleven major watersheds of Loudoun County. The relationship between urbanized area per watershed and mean fragment size showed a strong negative decay form (r ²=0.757, p<0.0001). Analysis of four landscape metrics showed increasing fragmentation of forest cover, particularly from 1987 to 1999, as well as an increase in forest edge and shape complexity. Of the landscape metrics used, the perimeter-to-area (P/A) ratio showed the strongest relationship with mean radiance of forest patches. In addition, there was a negative, linear relationship between distance from major roads and band 6 radiance of forested pixels. Overall, the study shows that landscape metrics can convey meaningful information on biophysical changes associated with forest fragmentation at broad scales. These changes suggest that ambient temperature increases associated with urban sprawl may have important, long-term implications for ecophysiological processes.     

The bovine neutrophil: Structure and function in blood and milk

Migration of polymorphonuclear neutrophil leukocytes (PMN) into the mammary gland provide the first line of defense against invading mastitis pathogens. Bacteria release potent toxins that activate white blood cells and epithelial cells in the mammary gland to secrete cytokines that recruit PMN that function as phagocytes at the site of infection. While freshly migrated PMN are active phagocytes, continued exposure of PMN to inhibitory factors in milk such as fat globules and casein, leads to altered PMN morphology and reduced phagocytosis. In the course of phagocytosing and destroying invading pathogens, PMN release chemicals that not only kill the pathogens but that also cause injury to the delicate lining of the mammary gland. This will result in permanent scarring and reduced numbers of milk secretory cells. The life span of PMN is limited by the onset of apoptosis. To minimize damage to mammary tissue, PMN undergo a specialized process of programmed cell death known as apoptosis. Macrophages quickly engulf and phagocytose apoptotic PMN, thereby minimizing the release of PMN granular contents that are damaging to tissue. The PMN possess an array of cell surface receptors that allow them to adhere and migrate through endothelium and to recognize and phagocytose bacteria. One receptor found on phagocytes that is receiving considerable attention in the control of infections by Gram-negative bacteria is CD14. Binding of lipopolysaccharide (LPS) to membrane bound CD14 causes release of tumor necrosis factor-alpha and sepsis. Binding of LPS to soluble CD14 shed from CD14-bearing cells results in neutralization of LPS and rapid recruitment of PMN to the site of infection. Recent advances in the fields of genomics and proteomics should greatly enhance our understanding of the PMN role in controlling intramammary infections in ruminants. Further, manipulation of PMN, through either recombinant proteins such as soluble CD14 that enhance PMN response or agents that mediate PMN apoptosis, may serve as novel therapeutics for the treatment of mastitis.