Dereplication Strategies for Targeted Isolation of New Antitrypanosomal Actinosporins A and B from a Marine Sponge Associated-Actinokineospora sp. EG49

High resolution Fourier transform mass spectrometry (HRFTMS) and nuclear magnetic resonance (NMR) spectroscopy were employed as complementary metabolomic tools to dereplicate the chemical profile of the new and antitrypanosomally active sponge-associated bacterium Actinokineospora sp. EG49 extract. Principal Component (PCA), hierarchical clustering (HCA), and orthogonal partial least square-discriminant analysis (OPLS-DA) were used to evaluate the HRFTMS and NMR data of crude extracts from four different fermentation approaches. Statistical analysis identified the best culture one-strain-many-compounds (OSMAC) condition and extraction procedure, which was used for the isolation of novel bioactive metabolites. As a result, two new O-glycosylated angucyclines, named actinosporins A (1) and B (2), were isolated from the broth culture of Actinokineospora sp. strain EG49, which was cultivated from the Red Sea sponge Spheciospongia vagabunda. The structures of actinosporins A and B were determined by 1D- and 2D-NMR techniques, as well as high resolution tandem mass spectrometry. Testing for antiparasitic properties showed that actinosporin A exhibited activity against Trypanosoma brucei brucei with an IC₅₀ value of 15 µM; however no activity was detected against Leishmania major and Plasmodium falciparum, therefore suggesting its selectivity against the parasite Trypanosoma brucei brucei; the causative agent of sleeping sickness.     

Hemodynamic signals correlate tightly with synchronized gamma oscillations

Functional imaging methods monitor neural activity by measuring hemodynamic signals. These are more closely related to local field potentials (LFPs) than to action potentials. We simultaneously recorded electrical and hemodynamic responses in the cat visual cortex. Increasing stimulus strength enhanced spiking activity, high-frequency LFP oscillations, and hemodynamic responses. With constant stimulus intensity, the hemodynamic response fluctuated; these fluctuations were only loosely related to action potential frequency but tightly correlated to the power of LFP oscillations in the gamma range. These oscillations increase with the synchrony of synaptic events, which suggests a close correlation between hemodynamic responses and neuronal synchronization.     

Development during storage of spraing symptoms in potato tubers infected with tobacco rattle virus

Summary In trials with potato tubers infected with tobacco rattle virus (TRV), symptoms of spraing in cvs Bellona, King Edward, Maris Bard, Matilda, Sv 82146 and Sv 82149 increased during storage when the tubers were cut. Storage of intact tubers at a constant temperature of 9°C or at fluctuating temperatures (2 weeks at 18°C, 2 weeks at 9°C and 2 weeks at 18°C) did not increase the frequency of symptoms.     

Rapid and widely disseminated acute phase protein response after experimental bacterial infection of pigs

The acute phase protein response is a well-described generalized early host response to tissue injury, inflammation and infection, observed as pronounced changes in the concentrations of a number of circulating serum proteins. The biological function of this response and its interplay with other parts of innate host defence reactions remain somewhat elusive. In order to gain new insight into this early host defence response in the context of bacterial infection we studied gene expression changes in peripheral lymphoid tissues as compared to hepatic expression changes, 14–18 h after lung infection in pigs. The lung infection was established with the pig specific respiratory pathogen Actinobacillus pleuropneumoniae. Quantitative real-time PCR based expression analysis were performed on samples from liver, tracheobronchial lymph node, tonsils, spleen and on blood leukocytes, supplemented with measurements of interleukin-6 and selected acute phase proteins in serum. C-reactive protein and serum amyloid A were clearly induced 14–18 h after infection. Extrahepatic expression of acute phase proteins was found to be dramatically altered as a result of the lung infection with an extrahepatic acute phase protein response occurring concomitantly with the hepatic response. This suggests that the acute phase protein response is a more disseminated systemic response than previously thought. The current study provides to our knowledge the first example of porcine extrahepatic expression and regulation of C-reactive protein, haptoglobin, fibrinogen, pig major acute phase protein, and transferrin in peripheral lymphoid tissues.     

Large-scale biotechnological production of the antileukemic marine natural product sorbicillactone A

In the search for novel bioactive compounds from sponge-derived microorganisms, we have recently identified two structurally and biosynthetically unprecedented fungal metabolites, the novel-type alkaloids sorbicillactone A and sorbicillactone B. Sorbicillactone A is active against leukemia cells without showing notable cytotoxicity. Therefore, we have developed an efficient process for its biotechnological production and isolation on a large scale supplying sufficient material for the ongoing preclinical investigations and structure-activity relationship (SAR) studies.     

Tentative Avenanthramide‐Modifying Enzyme in Oats

A decrease of the concentration of the synthetic avenanthramide N‐(4′‐hydroxy‐(E)‐cinnamoyl)‐5‐hydroxyanthranilic acid in a buffered slurry of milled oat groats (Avena sativa L.) was temperature and pH‐dependent, with a maximum rate at 30°C and pH 9. The reaction was inhibited in the presence of 2‐mercaptoethanol, acetic acid and at high temperature; suggestive of enzymatically catalyzed nature. Among eight different synthetic avenanthramides tested, the tentative enzyme had highest affinity for avenanthramides comprising caffeic and p‐coumaric acids and lowest for those comprising sinapic and ferulic acids. The activity was found in samples from several oat cultivars and was equally pronounced in both bran and endosperm flour of oats. Steeping of oat grains did not influence the reaction.     

Standardization of canine meningioma grading: Validation of new guidelines for reproducible histopathologic criteria

Canine meningiomas are currently graded using the human grading system. Recently published guidelines have adapted the human grading system for use in dogs. The goal of this study was to validate the new guidelines for canine meningiomas. To evaluate the inter-observer agreement, 5 veterinary surgical pathologists graded 158 canine meningiomas following the human grading system alone or with the new guidelines. The inter-observer agreement for histologic grade and each of the grading criteria (mitotic grade, invasion, spontaneous necrosis, macronucleoli, small cells, hypercellularity, pattern loss and anaplasia) was evaluated using the Fleiss kappa index. The diagnostic accuracy (sensitivity and specificity) was assessed by comparing the diagnoses obtained with the 2 grading systems with a consensus grade (considered the reference classification). The consensus histologic grade was obtained by agreement between 4 experienced veterinary neuropathologists following the guidelines. Compared with the human grading alone, the canine-specific guidelines increased the inter-observer agreement for: histologic grade (κ = 0.52); invasion (κ = 0.67); necrosis (κ = 0.62); small cells (κ = 0.36); pattern loss (κ = 0.49) and anaplasia (κ = 0.55). Mitotic grade agreement remained substantial (κ = 0.63). The guidelines improved the sensitivity in identifying grade 1 (95.6%) and the specificity in identifying grade 2 (96.2%) meningiomas. In conclusion, the new grading guidelines for canine meningiomas are associated with an overall improvement in the inter-observer agreement and higher diagnostic accuracy in diagnosing grade 1 and grade 2 meningiomas.     

Stability of Oat Avenanthramides

The three main oat avenanthramides, N‐(4′‐hydroxy)‐(E)‐cinnamoyl‐5‐hydroxyanthranilic acid ( Bp ), N‐(4′‐hydroxy‐3′‐methoxy)‐(E)‐cinnamoyl‐5‐hydroxyanthranilic acid ( Bf ), and N‐(3′,4′‐dihydroxy)‐(E)‐cinnamoyl‐5‐hydroxyanthranilic acid ( Bc ), and their corresponding cinnamic acids, p‐coumaric ( P ), ferulic ( F ), and caffeic ( C ), were investigated for stability to pH, temperature, and UV‐light treatment. The retention of the avenanthramides after processing of oat‐based food products was also analyzed. The avenanthramide Bc and the cinnamic acid C were sensitive to alkali and neutral conditions, especially in combination with heat treatment, whereas the other compounds studied were more stable. The cinnamic acids but not the avenanthramides were isomerized when irradiated with UV‐light. The avenanthramides were restored after processing of oat‐based products.