A class of potent antimalarials and their specific accumulation in infected erythrocytes

During asexual development within erythrocytes, malaria parasites synthesize considerable amounts of membrane. This activity provides an attractive target for chemotherapy because it is absent from mature erythrocytes. We found that compounds that inhibit phosphatidylcholine biosynthesis de novo from choline were potent antimalarial drugs. The lead compound, G25, potently inhibited in vitro growth of the human malaria parasites Plasmodium falciparum and P. vivax and was 1000-fold less toxic to mammalian cell lines. A radioactive derivative specifically accumulated in infected erythrocytes to levels several hundredfold higher than in the surrounding medium, and very low dose G25 therapy completely cured monkeys infected with P. falciparum and P. cynomolgi.     

Reproductive characteristics in female Swedish moose (Alces alces), with emphasis on puberty,timing of oestrus,and mating

Background

The moose (Alces alces) is an intensively managed keystone species in Fennoscandia. Several aspects of reproduction in moose have not been fully elucidated, including puberty, timing of mating and oestrus, and the length of the oestrus period. These aspects are relevant for an adaptive management of moose with respect to harvest, population size, demography and environmental conditions. Therefore, an investigation of female moose reproduction was conducted during the moose-hunting period in southern Sweden from 2008 to 2011.

Results

A total of 250 reproductive organs and information on carcass weight and age was collected from four different hunting areas (provinces of Öland, Småland, Södermanland, and Västergötland) in southern Sweden. The results showed that puberty in female moose varied with carcass weight, age, and time of season. The period for oestrous/mating lasted from about mid September to the beginning of November.

Conclusions

The oestrus period (predominantly for heifers) is longer than previously reported and was not finished when the hunting period started. Sampling the uterine cervix to detect spermatozoa was a useful method to determine if mating had occurred. To avoid hunting of moose during oestrus, we suggest that the hunting period should be postponed by at least 14 days in southern Sweden.     

Surveys on Coxiella burnetii infections in Swedish cattle,sheep, goats and moose

Background

Q fever is a zoonotic disease caused by the bacterium Coxiella burnetii. Prevalence data in ruminant species are important to support risk assessments regarding public and animal health. The aim was to investigate the presence of or exposure to C. burnetii in cattle, sheep, goats and moose, and to compare two enzyme-linked immunosorbent assays (ELISAs). National surveys of antibodies against C. burnetii were performed for dairy cattle (n=1537), dairy goats (n=58) and sheep (n=518). Bovine samples consisted of bulk milk, caprine of pooled milk, and ovine of pooled serum. Antibodies were investigated in moose samples (n=99) from three regions. A one-year regional cattle bulk milk survey was performed on the Isle of Gotland (n=119, four occasions). Cattle, sheep and goat samples were analysed with indirect ELISA and moose samples with complement fixation test. For the sheep, goat, and parts of the cattle survey, samples were run in parallel by ELISAs based on antigens from infected ruminants and ticks. Bulk milk samples from the regional cattle survey and vaginal swabs from a subset of the sheep herds (n=80) were analysed for the agent by polymerase chain reaction. Spatial clustering was investigated in the national cattle survey.

Results

The prevalence of antibodies in dairy herds was 8.2% with large regional differences. High risk clusters were identified in the southern regions. The prevalence among dairy herds on the Isle of Gotland varied from 55.9% to 64.6% and 46.4% to 58.9.0% for antibodies and agent, respectively, overall agreement between agent and antibodies was 85.2%. The prevalence of antibodies in sheep was 0.6%, the agent was not detected the vaginal swabs. Antibodies were not detected in goats or moose, although parts of the moose samples were collected in an area with high prevalence in cattle. The overall agreement between the two ELISAs was 90.4%.

Conclusions

The prevalence of antibodies against C. burnetii in dairy cattle in Sweden shows large regional differences. The results suggest that C. burnetii is a rare pathogen among Swedish moose, dairy goat and sheep. ELISAs based on ruminant and tick antigen performed in a similar manner under Swedish conditions.     

Isolation of horse mononuclear cells,especially of monocytes,on Isopaque-Ficoll neutral density gradient

Horse mononuclear cells were separated from whole blood using neutral density gradient centrifugation on Isopaque-Ficoll. The resulting cell suspension was comparable in composition with similarly prepared human and bovine mononuclear cell preparations. The relative concentration of monocytes was increased by the use of a gradient with density lower than that originally proposed by Böyum (Böyum, A. 1968. Scand. J. Clin. Lab. Investig. 21 supple. 97:77–89). Contamination by neutrophils was limited either by using a gradient medium of lower density or by replacing Isopaque-Ficoll by Percoll-0.9% NaCl. Although the density of the Isopaque-Ficoll appears to be the main determinant in the isolation method of Böyum, the mechanism of separation of the cell population is complex and a substantial variability of the results can be expected.     

Isozyme analysis of genetic diversity in wild Sicilian populations of Brassica sect. Brassica in view of genetic resources management

In Sicily and in the small surrounding islands the section Brassica of the genus Brassica comprises five species, B. insularis Moris, B. incana Ten., B. macrocarpa Guss., B. rupestris Raf. and B. villosa Biv. These taxa represent a genetic resource as relatives of kale crops but some populations are endangered or threatened, thus isozyme analyses were performed to assess the genetic diversity degree at population and species levels in order to assist the design of conservation management programs.Eleven loci from five enzyme systems (aconitase, leucine aminopeptidase, 6-phosphogluconate dehydrogenase, phosphoglucoisomerase phosphoglucomutase) were analyzed in sixteen natural population (fifteen from Sicily, one from Calabria). Mean within-population genetic diversity was moderate (P = 41%, A = 1.54, H = 0.16). In some cases a great number of heterozygous individuals were detected, in other cases fixation index (F) deviated significantly from Hardy-Weinberg genotypic expectations.A total of 37 alleles was recognized, six of which resulted exclusive to single populations. The among-population component of the total genetic diversity (Gst mean values) for each species was 0.30–0.37, indicating genetic differentiation among populations.Among B. villosa and B. rupestris populations genetic distance values resulted rather low and they resulted high with B. incana and B. macrocarpa populations.The results are discussed with regard to the distribution of the genetic diversity level and the genetic resources management.     

Effects of garlic powders with varying alliin contents on hepatic drug metabolizing enzymes in rats

The anticarcinogenic effect of garlic has been demonstrated in both epidemiologic and experimental studies. In this study, possible mechanisms involved in the anticarcinogenic effect of garlic consumption were assessed by determining its capacity to alter drug metabolizing enzymes, in relation with its alliin content. Rats were fed a diet for 2 weeks containing 5% garlic powders produced from bulbs grown on soils with different levels of sulfate fertilization and therefore containing differing amounts of alliin. Activities of several hepatic enzymes, which are important in carcinogen metabolism such cytochromes P450 (CYP) and phase II enzymes, were determined. Garlic consumption slightly increased ethoxyresorufin O-deethylase and CYP 1A2 levels. In contrast, garlic consumption decreased CYP 2E1 activity and the level of the corresponding isoform. UDP glucuronosyl transferase and glutathion S-transferase activities were increased by garlic powders. The alliin content of the garlic powders was positively correlated with UGT activity although not with other activities. Effects produced by garlic consumption were qualitatively similar to that of diallyl disulfide, a sulfur compound that has been extensively studied. These data could partially explain the chemoprotective effect of garlic.     

Intraobserver and interobserver repeatability of ocular biometric measurements obtained by means of B-mode ultrasonography in dogs

OBJECTIVE: To assess intra- and interobserver repeatability of ocular biometric measurements obtained by means of high-resolution B-mode ultrasonography in dogs. Animals-6 Beagles without ocular abnormalities. PROCEDURES: B-mode ultrasonography was performed bilaterally with a 10.5-MHz broadband compact linear array transducer. All measurements were made on 2 different occasions by 2 observers. The Bland-Altman method was used to assess agreement between measurements obtained by the 2 observers and between the 2 sets of measurements obtained by each observer. RESULTS: Intra- and interobserver repeatability was highest for larger measurements, such as depth of the eye and depth of the anterior chamber. When repeatability was examined, bias was significantly different from 0 for only a few measurements, but the percentage difference between observations was as high as 180% for some measurements. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that most measurements of intraocular distances and structures obtained by means of high-resolution B-mode ultrasonography have acceptable intra- and interobserver repeatability. However, the percentage difference between observations can be high for smaller measurements.     

Low-Molecular-Weight Fucoidan Induces Endothelial Cell Migration via the PI3K/AKT Pathway and Modulates the Transcription of Genes Involved in Angiogenesis

Low-molecular-weight fucoidan (LMWF) is a sulfated polysaccharide extracted from brown seaweed that presents antithrombotic and pro-angiogenic properties. However, its mechanism of action is not well-characterized. Here, we studied the effects of LMWF on cell signaling and whole genome expression in human umbilical vein endothelial cells and endothelial colony forming cells. We observed that LMWF and vascular endothelial growth factor had synergistic effects on cell signaling, and more interestingly that LMWF by itself, in the absence of other growth factors, was able to trigger the activation of the PI3K/AKT pathway, which plays a crucial role in angiogenesis and vasculogenesis. We also observed that the effects of LMWF on cell migration were PI3K/AKT-dependent and that LMWF modulated the expression of genes involved at different levels of the neovessel formation process, such as cell migration and cytoskeleton organization, cell mobilization and homing. This provides a better understanding of LMWF’s mechanism of action and confirms that it could be an interesting therapeutic approach for vascular repair.