1-O-Alkylglycerol Ethers from the Marine Sponge Guitarra abbotti and Their Cytotoxic Activity

The cytotoxicity-bioassay-guided fractionation of the ethanol extract from the marine sponge Guitarra abbotti, whose 1-O-alkyl-sn-glycerol ethers (AGEs) have not been investigated so far, led to the isolation of a complex lipid fraction containing, along with previously known compounds, six new lipids of the AGE type. The composition of the AGE fraction as well as the structures of 6 new and 22 previously known compounds were established using ¹H and ¹³C NMR, GC/MS, and chemical conversion methods. The new AGEs were identified as: 1-O-(Z-docos-15-enyl)-sn-glycerol (1), 1-O-(Z-docos-17-enyl)-sn-glycerol (2), 1-O-(Z-tricos-15-enyl)-sn-glycerol (3), 1-O-(Z-tricos-16-enyl)-sn-glycerol (4), 1-O-(Z-tricos-17-enyl)-sn-glycerol (5), and 1-O-(Z-tetracos-15-enyl)-sn-glycerol (6). The isolated AGEs show weak cytotoxic activity in THP-1, HL-60, HeLa, DLD-1, SNU C4, SK-MEL-28, and MDA-MB-231 human cancer cells. A further cytotoxicity analysis in JB6 P⁺ Cl41 cells bearing mutated MAP kinase genes revealed that ERK2 and JNK1 play a cytoprotective role in the cellular response to the AGE-induced cytotoxic effects.     

接种茄病镰刀菌(Fusarium solani f.sp.glycines)对不同基因型大豆植株和须根的影响

摘要：茄病镰刀菌（Fusarium solani f．sp．glydne）是一种土生细菌,通过侵染大豆根系引发猝死综合症。利用温室盆栽试验和须根培养试验研究了接种茄孢镰刀菌对13个不同基因型大豆的影响。结果表明：接种后所有盆栽大豆主根都有明显深褐色的侵染病斑;移植后21d测定了盆栽植株叶部发病程度,Peking表现最为严重,然后依次为Spencer,Ripley,P3981,Williams82,Essex,Forrest,Iroquois,PI520733,Hartwig,PI567650B,Jack,和PI567374。叶部发病程度与冠高（r=-0．422,P=0．0018）、冠重（r=-0．857,P〈0．0001）和根重（r=-0．732,P〈0．0001）呈显著负相关。主根病斑长度与叶部发病程度没有相关性,表明大豆对病原菌的抗性不能仅通过根系得到充分控制。对培养的大豆须根接种茄病镰刀菌菌丝体10d后,不同基因型大豆的菌落直径的变化范围为17—40mm,差异显著（P=0．05）,其中Spencer和Peking须根上的菌落直径显著（P=0．05）大于PI567374和PI520733。对Spencer和PI567374的须根接种10灿茄病镰刀菌常量成分悬浮液,10d后菌落直径分别为50和38mm,差异显著（P=0．05）。通常,不同基因型大豆间茄病镰刀菌在培养须根上的生长与整株的症状间有一定的相关性,但不总是这样,这是因为即使根系对毒素产生抗性来减少叶部病害症状,但并不是所有的大豆都表现出明显的根系抗性。     

Giant cell tumor of the bone in a scarlet macaw (Ara macao)

A 6-mo-old female scarlet macaw (Ara macao) was presented after a 2-mo period of anorexia and weakness. The bird was reluctant to fly 1 wk before referral due to a painful left wing. Physical examination revealed a firm swelling around the left shoulder. On radiographs, the diaphysis and proximal metaphysis of the left scapula were radiolucent. Computer tomography revealed an osteolytic process, suggestive of a bone tumor, affecting the left scapula. Cytology of a fine needle aspiration biopsy of the mass showed erythrocytes, a proliferation of spindle-shaped mesenchymal cells, and multinucleated giant cells (osteoclasts) suggestive of a giant cell tumor. The left wing, including the scapula, was amputated. The bird showed a fast recovery but died 1 hr later. Findings during the pathological examination were compatible with shock due to blood loss. The shoulder process was characterized as a giant cell tumor. To our knowledge, this is the first complete report of a giant cell tumor of the bone in a bird.     

Diagnostic and public health investigation of Mycobacterium tuberculosis infection in a dog in Ontario,Canada

A 4-y-old, female mixed-breed dog was presented to the Ontario Veterinary College for further evaluation of multiple pulmonary and hepatic masses, intrathoracic lymphadenitis, and recent development of a pyogranulomatous pleural effusion. Along with other comprehensive tests, a thoracic lymph node biopsy was performed, and Mycobacterium tuberculosis complex infection was confirmed by real-time PCR. The dog’s condition declined post-operatively, and euthanasia was elected. Postmortem examination confirmed severe granulomatous pneumonia, hepatitis, intrathoracic and intraabdominal lymphadenitis, omentitis, and nephritis. Line-probe assays performed on samples collected postmortem confirmed the species as M. tuberculosis. 24-loci MIRU-VNTR genotyping, spoligotyping, and whole-genome sequencing revealed relations to known human isolates, but no epidemiologic link to these cases was investigated. Given the concern for potential human exposure during this animal’s disease course, a public health investigation was initiated; 45 individuals were tested for M. tuberculosis exposure, and no subsequent human infections related to this animal were identified. Our case highlights the need for more readily available, minimally invasive testing for the diagnosis of canine mycobacteriosis, and highlights the ability of canid species to act as potential contributors to the epidemiology of M. tuberculosis infections.     

Influence of the Structural Features of Carrageenans from Red Algae of the Far Eastern Seas on Their Antiviral Properties

The structural diversity and unique physicochemical properties of sulphated polysaccharides of red algae carrageenans (CRGs), to a great extent, determine the wide range of their antiviral properties. This work aimed to compare the antiviral activities of different structural types of CRGs: against herpes simplex virus type 1 (HSV-1) and enterovirus (ECHO-1). We found that CRGs significantly increased the resistance of Vero cells to virus infection (preventive effect), directly affected virus particles (virucidal effect), inhibited the attachment and penetration of virus to cells, and were more effective against HSV-1. CRG1 showed the highest virucidal effect on HSV-1 particles with a selective index (SI) of 100. CRG2 exhibited the highest antiviral activity by inhibiting HSV-1 and ECHO-1 plaque formation, with a SI of 110 and 59, respectively, when it was added before virus infection. CRG2 also significantly reduced the attachment of HSV-1 and ECHO-1 to cells compared to other CRGs. It was shown by molecular docking that tetrasaccharides—CRGs are able to bind with the HSV-1 surface glycoprotein, gD, to prevent virus–cell interactions. The revealed differences in the effect of CRGs on different stages of the lifecycle of the viruses are apparently related to the structural features of the investigated compounds.     

Evaluation of single and comparative intradermal tuberculin tests for tuberculosis eradication in caprine flocks in Castilla y León (Spain)

Goats can act as reservoirs for tuberculosis (TB) infection. The main etiological agents of TB in goats are Mycobacterium caprae and Mycobacterium bovis and they infect also a wide range of domestic and wild animals and humans. Control programmes based mainly on the application of single and comparative intradermal tuberculin (SIT and SCIT respectively) tests are being implemented in certain regions of Spain with a high density of caprine flocks as Castilla y León, including goats with epidemiological relationship with cattle. The aim of this study was to evaluate the performance of the intradermal tests in naturally TB-infected caprine flocks from this region. The study was performed using data from 17,450 goats in 54 different flocks that were classified as TB-infected in the control programmes executed in 2010 and 2011. Data from 1237 goats from 7 dairy flocks depopulated after the first intradermal testing were used to estimate the sensitivity (Se) using bacteriology as the gold-standard. Overall Se of the SIT test using the severe interpretation was 43.9% (CI 95%, 40.4–47.4) and decreased to 38.8% (CI 95%, 35.5–42.3) using the standard interpretation. Overall Se of the SCIT test ranged between 21.3% (CI 95%, 17.6–25.4) and 7% (CI 95%, 4.9–9.8) depending of the interpretation criteria. A significant weak positive correlation was found between age and skin fold thickness (Spearman’s test p < 0.05). Results from this study yielded, in general, low Se values probably due the systematic detection and slaughter of reactors as a consequence of the eradication programme in previous years or the presence of factors that may interfere in the diagnosis. Therefore, these results suggest the necessity of including ancillary diagnostic tools and/or strict interpretation criteria to maximize detection of positive animals in infected settings.     

Morphological characteristics of blood cells in monitor lizards: is erythrocyte size linked to actual body size?

Blood cell morphology and count are not uniform across species. Recently, between‐species comparisons revealed that the size of red blood cells is associated with body size in some lizard taxa, and this finding was interpreted in the context of the metabolic theory. In the present study, we examined the numbers and the size of blood cells in 2 species of monitor lizards, the mangrove‐dwelling monitor (Varanus indicus) and the savannah monitor (V. exanthematicus), and we compared these traits in individuals of different body size. The results revealed that during the course of ontogeny, the size of red blood cells increases with body mass. Because the mass‐specific metabolic rate decreases with body size and the cell volume‐to‐surface ratio decreases with the cell size, changes in the erythrocyte size might be the result of oxygen transport adjustment.     

The influence of topography and fire in controlling landscape composition and structure in Sierra de Gredos (Central Spain)

Mediterranean landscapes are dynamic systems that undergo temporal changes in composition and structure in response to disturbances, such as fire. Neither landscape patterns nor driving factors that affect them are evenly distributed in space. Accordingly, disturbances and biophysical factors interact in space through time. The aim of this paper is to assess the relative influence of topography and fire on the landscape patterns of a large forested area located in Sierra de Gredos (Central Spain) through time. A series of Landsat MSS images from 1975 to 1990, and a digital elevation model (DEM) were used to map fires, assess topographical complexity and evaluate changes in landscape composition and structure. Functional regions across the entire landscape were identified using different classification criteria (i.e., percentage burned area and topographic properties) to model topographic and fire impacts at regional scales. A canonical variance partition method, with a time series split-plot design, quantified the relative influence and co-variation of topography and fire on land cover patterns through time. Main results indicated that analyzing portions of the landscape under similar environmental conditions and fire histories, the effects of different fire regimes on the spatio-temporal dynamics of main land covers can be highlighted. However, the impact of fire on landscape patterns was high variable among regions due to the different regeneration abilities of main land covers, the topographic constraints and the fire histories of each region. Hence, broad patterns of fire related variance and co-variation with topography emerged across the entire area due to the different conditions of each landscape portion in which this large Mediterranean landscape was divided. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Trichoderma Biocontrol of Colletotrichum acutatum and Botrytis cinerea and Survival in Strawberry

Trichoderma isolates are known for their ability to control plant pathogens. It has been shown that various isolates of Trichoderma, including T. harzianum isolate T-39 from the commercial biological control product TRICHODEX, were effective in controlling anthracnose (Colletotrichum acutatum) and grey mould (Botrytis cinerea) in strawberry, under controlled and greenhouse conditions. Three selected Trichoderma strains, namely T-39, T-161 and T-166, were evaluated in large-scale experiments using different timing application and dosage rates for reduction of strawberry anthracnose and grey mould. All possible combinations of single, double or triple mixtures of Trichoderma strains, applied at 0.4% and 0.8% concentrations, and at 7 or 10 day intervals, resulted in reduction of anthracnose severity; the higher concentration (0.8%) was superior in control whether used with single isolates or as a result of combined application of two isolates, each at 0.4%. Only a few treatments resulted in significant control of grey mould. Isolates T-39 applied at 0.4% at 2 day intervals, T-166 at 0.4%, or T-161 combined with T-39 at 0.4% were as effective as the chemical fungicide fenhexamide. The survival dynamics of populations of the Trichoderma isolates (T-39, T-105, T-161 and T-166) applied separately was determined by dilution plating and isolates in the mixtures calculated according to the polymerase chain reaction (PCR) using repeat motif primers. The biocontrol isolates were identified to the respective species T. harzianum (T-39), T. hamatum (T-105), T. atroviride (T-161) and T. longibrachiatum (T-166), according to internal transcribed spacer sequence analysis.     

Identification of the molecular make-up of the Potato virus Y strain PVY(Z): genetic typing of PVY(Z)-NTN

Potato virus Y (PVY) strains were originally defined by interactions with different resistance genes in standard potato cultivars. Five distinct strain groups are defined that cause local or systemic hypersensitive responses (HRs) in genetic background with a corresponding N gene: PVY(O), PVY(N), PVY(C), PVY(Z), and PVY(E). The nucleotide sequences of multiple isolates of PVY(O) and PVY(N) differ from each other by ≈8% along their genomes. Additionally, complete genome sequences of multiple recombinant isolates are composed of segments of parental PVY(O) and PVY(N) sequences. Here, we report that recombinant isolate PVY-L26 induces an HR in potato 'Maris Bard' carrying the putative Nz gene, and is not recognized by two other resistance genes, Nc and Ny(tbr). These genetic responses in potato, combined with the inability of PVY-L26 to induce vein necrosis in tobacco, clearly define it as an isolate from the PVY(Z) strain group and provide the first information on genome structure and sequence of PVY(Z). The genome of PVY-L26 displays typical features of European NTN-type isolates with three recombinant junctions (PVY(EU-NTN)), and the PVY-L26 is named PVY(Z)-NTN. Three typical PVY(NTN) isolates and two PVY(N) isolates, all inducing vein necrosis in tobacco, were compared with PVY-L26. One PVY(NTN) isolate elicited HR reactions in Maris Bard, similar to PVY-L26, while two induced a severe systemic HR-like reaction quite different from the quasi-symptomless reaction induced by two PVY(N) isolates. 'Yukon Gold' potato from North America produced HR against several PVY(NTN) isolates, including PVY-L26, but only late and limited systemic necrosis against one PVY(N) isolate. Consequently, according to symptoms in potato indicators, both PVY(Z) and PVY(NTN) isolates appeared biologically very close and clearly distinct from PVY(O) and PVY(N) strain groups.