Analysis of Mycosporine-Like Amino Acids in Selected Algae and Cyanobacteria by Hydrophilic Interaction Liquid Chromatography and a Novel MAA from the Red Alga Catenella repens

Mycosporine-like amino acids (MAAs), a group of small secondary metabolites found in algae, cyanobacteria, lichens and fungi, have become ecologically and pharmacologically relevant because of their pronounced UV-absorbing and photo-protective potential. Their analytical characterization is generally achieved by reversed phase HPLC and the compounds are often quantified based on molar extinction coefficients. As an alternative approach, in our study a fully validated hydrophilic interaction liquid chromatography (HILIC) method is presented. It enables the precise quantification of several analytes with adequate retention times in a single run, and can be coupled directly to MS. Excellent linear correlation coefficients (R² > 0.9991) were obtained, with limit of detection (LOD) values ranging from 0.16 to 0.43 µg/mL. Furthermore, the assay was found to be accurate (recovery rates from 89.8% to 104.1%) and precise (intra-day precision: 5.6%, inter-day precision ≤6.6%). Several algae were assayed for their content of known MAAs like porphyra-334, shinorine, and palythine. Liquid chromatography-mass spectrometry (LC-MS) data indicated a novel compound in some of them, which could be isolated from the marine species Catenella repens and structurally elucidated by nuclear magnetic resonance spectroscopy (NMR) as (E)-3-hydroxy-2-((5-hydroxy-5-(hydroxymethyl)-2-methoxy-3-((2-sulfoethyl)amino)cyclohex-2-en-1-ylidene)amino) propanoic acid, a novel MAA called catenelline.     

The porcine immune system--differences compared to man and mouse and possible consequences for infections by Salmonella serovars

The porcine immune system differs in many aspects from that of humans and mice. Morphological differences in the lymphatic system (e.g. lymph nodes, Peyer's patches), and phenotypic differences in immune cells have been observed as well as functional differences in immune cell populations. Indeed, even the prenatal development of the embryonic piglet proceeds in a principally different way. However, it remains unclear to what extent these differences might contribute to the predisposition to and outcomes of bacterial infections, in particular those with zoonotic potential. In the following article we will review some of the peculiarities of the porcine immune system and consider possible implications for the course of infections in swine, with an emphasis on Salmonella serovars.     

Schiff-Sherrington syndrome in a 7-year-old zebra mare (Equus quagga)

A 7-year-old plains zebra (Equus quagga) mare was presented to the Clinic for Horses, University of Veterinary Medicine Hannover, Foundation, Germany as an emergency. The owner reported that the mare was unable to stand after a suspected traumatic episode at the home yard. The mare was transported in lateral recumbency in a trailer. Clinical examination revealed tachycardia (up to 102 beats/min), tachypnoea (28 breaths/min), mild fever (38.7°C) and clinical dehydration with elastic skin and dry and red mucous membranes. Neurological examination showed paraplegia with complete loss of nociception of the mid-thoracic region and caudal to it. In contrast, examination of the thoracic limbs revealed marked extensor rigidity and hyperaesthesia resulting in an enhanced acute response to minimal touch of the neck and shoulder region. Lateral radiographs of the thoracic vertebral column led to the diagnosis of a complete fracture of the vertebral body of the 7th thoracic (T) vertebra with suspected traumatic compression of the spinal cord. Post-mortem examination confirmed the diagnosis. Gross pathology revealed a complete comminuted fracture of T7 with marked compression of the spinal cord. This is the first report describing the clinical, radiological and pathological findings of a fracture of a thoracic vertebra leading to a Schiff-Sherrington syndrome in a zebra (Equus quagga).     

Stabilization mechanisms of organic matter in four temperate soils: Development and application of a conceptual model

Based on recent findings in the literature, we developed a process‐oriented conceptual model that integrates all three process groups of organic matter (OM) stabilization in soils namely (1) selective preservation of recalcitrant compounds, (2) spatial inaccessibility to decomposer organisms, and (3) interactions of OM with minerals and metal ions. The model concept relates the diverse stabilization mechanisms to active, intermediate, and passive pools. The formation of the passive pool is regarded as hierarchical structured co‐action of various processes that are active under specific pedogenetic conditions. To evaluate the model, we used data of pool sizes and turnover times of soil OM fractions from horizons of two acid forest and two agricultural soils. Selective preservation of recalcitrant compounds is relevant in the active pool and particularly in soil horizons with high C contents. Biogenic aggregation preserves OM in the intermediate pool and is limited to topsoil horizons. Spatial inaccessibility due to the occlusion of OM in clay microstructures and due to the formation of hydrophobic surfaces stabilizes OM in the passive pool. If present, charcoal contributes to the passive pool mainly in topsoil horizons. The importance of organo‐mineral interactions for OM stabilization in the passive pool is well‐known and increases with soil depth. Hydrophobicity is particularly relevant in acid soils and in soils with considerable inputs of charcoal. We conclude that the stabilization potentials of soils are site‐ and horizon‐specific. Furthermore, management affects key stabilization mechanisms. Tillage increases the importance of organo‐mineral interactions for OM stabilization, and in Ap horizons with high microbial activity and C turnover, organo‐mineral interactions can contribute to OM stabilization in the intermediate pool. The application of our model showed that we need a better understanding of processes causing spatial inaccessibility of OM to decomposers in the passive pool.     

Classification of cassava into ‘bitter’ and ‘cool’ in Malawi: From farmers' perception to characterisation by molecular markers

Cassava roots, a major food in Africa, contain cyanogenic glucosides that may cause toxic effects. Malawian women farmers considered fields of seemingly similar cassava plants to be mixes of both ‘cool’ and ‘bitter’ cultivars. They regard roots from ‘cool’ cultivars as non-toxic. Roots of ‘bitter’ were considered to require extensive traditional processing done by women to be safe for consumption. But curiously, these women farmers preferred ‘bitter’ cultivars since toxicity confers protection against theft, which was a serious threat to the food security of their families. We studied how well these farmers comprehend the effects of genetic variations in cassava when dealing with cyanogenesis in this complex system. Using molecular markers we show that most plants farmers identified as belonging to a particular named cultivar had a genotype typical of that cultivar. Farmers' ethno-classification into ‘cool’ and ‘bitter’ cultivars corresponded to a genetic sub-division of the typical genotypes of the most common cultivars, with four-fold higher cyanogenic glucoside levels in the bitter cultivars. Examining morphology, farmers distinguished genotypes better than did the investigators when using a standard botanical key. Undoubtedly, these women farmers grasp sufficiently the genetic diversity of cassava with regard to cyanogenesis to simultaneously benefit from it and avoid its dangers. Consequently, acyanogenic cassava – the breeding of which is an announced good of some cassava genetic improvement programmes – is not a priority to these farmers. Advances in molecular genetics can help improve food supply in Africa by rapid micropropagation, marker assisted breeding and introduction of transgenic varieties, but can also help to elucidate tropical small-scale farmers' needs and skills. This revised version was published online in August 2006 with corrections to the Cover Date.     

Intermediate landscape disturbance maximizes metapopulation density

The viability of metapopulations in fragmented landscapes has become a central theme in conservation biology. Landscape fragmentation is increasingly recognized as a dynamical process: in many situations, the quality of local habitats must be expected to undergo continual changes. Here we assess the implications of such recurrent local disturbances for the equilibrium density of metapopulations. Using a spatially explicit lattice model in which the considered metapopulation as well as the underlying landscape pattern change dynamically, we show that equilibrium metapopulation density is maximized at intermediate frequencies of local landscape disturbance. On both sides around this maximum, the metapopulation may go extinct. We show how the position and shape of the intermediate viability maximum is responding to changes in the landscape’s overall habitat quality and the population’s propensity for local extinction. We interpret our findings in terms of a dual effect of intensified landscape disturbances, which on the one hand exterminate local populations and on the other hand enhance a metapopulation’s capacity for spreading between habitat clusters.     

Structure determination of partially deuterated carotenoids from intrinsically labeled vegetables by HPLC-MS and 1H NMR

The structures of biosynthetic deuterated carotenoids in labeled vegetables were investigated: (all-E)-lutein and (all-E)-beta-carotene from spinach, and (all-E)-beta-carotene and (all-E)-alpha-carotene from carrots. The vegetables were grown hydroponically using a nutrient solution enriched with deuterium oxide (D(2)O) and were extracted using matrix solid-phase dispersion (MSPD). Deuterium enrichment in the carotenoid molecules was determined by liquid chromatography-mass spectrometry (LC-MS). (all-E)-Lutein and (all-E)-beta-carotene in spinach showed partial deuteration from (2)H(1) to (2)H(12), with the abundance maximum at (2)H(5). (all-E)-beta-Carotene and (all-E)-alpha-carotene from carrots showed partial deuteration from (2)H(1) to (2)H(17), with the abundance maximum at (2)H(11). The (1)H NMR spectra of the four deuterated carotenoids showed additional signals for all methyl groups and decreased signal intensity for the olefinic protons and the methylene protons in the ring. These differences are due to isotopic effects and are based on the substitution of protons by deuterium atoms. The deuteration was distributed randomly throughout the carotenoid molecules.     

Molecular mimicry regulates ABA signaling by SnRK2 kinases and PP2C phosphatases

Abscisic acid (ABA) is an essential hormone for plants to survive environmental stresses. At the center of the ABA signaling network is a subfamily of type 2C protein phosphatases (PP2Cs), which form exclusive interactions with ABA receptors and subfamily 2 Snfl-related kinase (SnRK2s). Here, we report a SnRK2-PP2C complex structure, which reveals marked similarity in PP2C recognition by SnRK2 and ABA receptors. In the complex, the kinase activation loop docks into the active site of PP2C, while the conserved ABA-sensing tryptophan of PP2C inserts into the kinase catalytic cleft, thus mimicking receptor-PP2C interactions. These structural results provide a simple mechanism that directly couples ABA binding to SnRK2 kinase activation and highlight a new paradigm of kinase-phosphatase regulation through mutual packing of their catalytic sites.