A new structural form in the SAM/metal-dependent o‑methyltransferase family: MycE from the mycinamicin biosynthetic pathway

O-linked methylation of sugar substituents is a common modification in the biosynthesis of many natural products and is catalyzed by multiple families of S-adenosyl-l-methionine (SAM or AdoMet)-dependent methyltransferases (MTs). Mycinamicins, potent antibiotics from Micromonospora griseorubida, can be methylated at two positions on a 6-deoxyallose substituent. The first methylation is catalyzed by MycE, a SAM- and metal-dependent MT. Crystal structures were determined for MycE bound to the product S-adenosyl-l-homocysteine (AdoHcy) and magnesium, both with and without the natural substrate mycinamicin VI. This represents the first structure of a natural product sugar MT in complex with its natural substrate. MycE is a tetramer of a two-domain polypeptide, comprising a C-terminal catalytic MT domain and an N-terminal auxiliary domain, which is important for quaternary assembly and for substrate binding. The symmetric MycE tetramer has a novel MT organization in which each of the four active sites is formed at the junction of three monomers within the tetramer. The active-site structure supports a mechanism in which a conserved histidine acts as a general base, and the metal ion helps to position the methyl acceptor and to stabilize a hydroxylate intermediate. A conserved tyrosine is suggested to support activity through interactions with the transferred methyl group from the SAM methyl donor. The structure of the free enzyme reveals a dramatic order-disorder transition in the active site relative to the S-adenosyl-l-homocysteine complexes, suggesting a mechanism for product/substrate exchange through concerted movement of five loops and the polypeptide C-terminus.     

Advances in the timing of spring cleaning by the honeybee Apis mellifera in Poland

1. The honeybee Apis mellifera is of huge worldwide economic importance in the pollination of crops for human consumption. In recent years, honeybee populations have declined under pressure from diseases and pests. Climate change is increasingly being viewed as an additional threat to honeybees and yet only limited research has been carried out in this area. 2. This paper reports the advance of the first cleansing flight (‘spring cleaning’) of the honeybee in Poznań, Poland, i.e. flights to excrete faeces, over a month in the period 1985–2009. The timing of this flight is advanced not only by higher late winter/spring temperatures but also by higher temperatures in the previous summer and autumn. 3. This earlier activity gives hope that the reported earlier flowering of many native and cultivated species will not cause a pollination synchrony crisis.     

Anatomy of the coracoid and diversity of the Procellariiformes (Aves) in the Oligocene of Europe

Abstract: Two European species of the Diomedeoididae, an extinct family of procellariiform (tube‐nose) birds, have hitherto been distinguished primarily by size of their limb bones. Here, we describe an Early Oligocene (Rupelian) procellariifom coracoid that in all probability represents the larger species, Diomedeoides lipsiensis, and compare it to the coracoids of smaller diomedeoidids and extant procellariiforms. Using multivariate (Principal Component Analysis) and univariate analyses, we demonstrate that nearly all measurements are heavily size dependent, which makes the proportions and some other shape characters of little use as phylogenetic markers. Among eight measurements, the coracoid corpus width shows the highest correlation (higher than corpus depth) with body mass, permitting a precise calculation of over twofold difference in body mass between D. lipsiensis and smaller species. Among 16 qualitative characters analysed, the majority proved too variable to be used as markers of interfamily relationships and only 2–3, the ventral intermuscular line, sternocoracoid articulation (divided vs. undivided), and, with reservations, epimarginal crest vary consistently between the families. By far the most variable is the acrocoracoid process that tends to be deeper (more elongate dorsoventrally) in larger petrels but not in the albatrosses. However, the detailed shapes of the acrocoracoid heads are highly genus specific and suggest a genus‐level diversity among the Diomedeoididae from the Oligocene of Europe. The common features of the diomedeoidid coracoids are best interpreted as plesiomorphies, which accounts for some similarities to the Oceanitinae (that are probably basal among the crown‐group procellariiforms). The evidence from the coracoid is consistent with a stem‐group position of the Diomedeoididae as previously proposed by others. We emphasize the need of a group‐specific character analysis, primarily of allometries and levels of character variation, prior to a phylogenetic reconstruction.     

Primary coronary intervention for ST-elevation myocardial infarction in Indonesia and the Netherlands: a comparison

Background. Although the beneficial effects of primary percutaneous coronary intervention (PCI) for ST-elevation myocardial infarction (STEMI) have been demonstrated in a number of trials, most studies were conducted in Western countries. Experience, logistics and patient characteristics may differ in other parts of the world. Methods. Consecutive patients treated with primary PCI in Cinere Hospital, Jakarta, Indonesia, between January 2008 and October 2008 were compared with those treated in the Isala Clinics, Zwolle, the Netherlands. Results. During the study period, a total of 596 patients were treated by primary PCI, 568 in Zwolle and 28 in Jakarta. Patients in Indonesia were younger (54 vs 63 years), more often had diabetes (36 vs. 12%) and high lipids and were more often smokers (68 vs. 31%). Time delay between symptom onset and admission was longer in Indonesia. Patients from Indonesia more often had signs of heart failure at admission. The time between admission and balloon inflation was longer in Indonesia. At angiography, patients from Indonesia more often had multivessel disease. There was no difference in the percentage of restoration of TIMI 3 flow by primary PCI between the two hospitals. Conclusion. Patients with STEMI in Indonesia have a higher risk profile compared with those in the Netherlands, according to prevalence of coronary risk factors, signs of heart failure, multivessel disease and patient delay. Time delay between admission and balloon inflation was much longer in Indonesia, because of both logistic and financial reasons. (Neth Heart J 2009;17:418–21.)     

Efficient hydrogen production from the lignocellulosic energy crop Miscanthus by the extreme thermophilic bacteria Caldicellulosiruptor saccharolyticus and Thermotoga neapolitana

Background  

The production of hydrogen from biomass by fermentation is one of the routes that can contribute to a future sustainable hydrogen economy. Lignocellulosic biomass is an attractive feedstock because of its abundance, low production costs and high polysaccharide content.     

A morphological and genetic analysis of the European bitterling species complex

Bitterling fishes lay their eggs on the gills of living freshwater mussels and are valuable models in behavioural and evolutionary ecology. We used morphological and genetic data to resolve the taxonomic relationships of bitterling in Europe. Previous studies have suggested the European bitterling is either a single species with a wide but discontinuous geographic distribution, or a complex of species. Morphometric and meristic data identified differences between three putative species; with a clear distinction between the eastern Asian Rhodeus sericeus, western European bitterling Rhodeus amarus, and colchian bitterling, Rhodeus colchicus. Polymorphism in the mitochondrial DNA control region was predominantly due to insertion/deletion events, making phylogenetic inference difficult, but the single haplotype found in R. sericeus populations was detected at low frequency (one of 24 individuals) in R. amarus and R. colchicus populations. Eight control region haplotypes were found in R. amarus populations, which were distinct from the two haplotypes in a R. colchicus population. Cytochrome b data produced a phylogeny with strongly‐supported differentiation between a clade of two R. sericeus haplotypes and a clade of six R. amarus/colchicus haplotypes. The star‐like topology of the R. amarus/colchicus haplotypes in a minimum spanning network suggested a rapid radiation in this clade. Our results are consistent with an hypothesis of relatively ancient divergence of R. sericeus from R. amarus/colchicus and more recent and rapid differentiation between R. amarus and R. colchicus. © 2008 The Linnean Society of London, Biological Journal of the Linnean Society, 2008, 95 , 337–347.     

Influence of induction of parturition on the neonatal acute phase response in foals

The objectives of the present study were to determine whether induction of parturition in mares at term with low doses of oxytocin (2.5 i.u. i.v. every 20 min) affected the incidence of peri-partum complications or inflammatory responses in the neonatal foal. Parturition was induced in 11 of 26 mares and the remainder foaled spontaneously. Serum concentrations of amyloid A (AA; an acute phase protein) were measured (with a commercial ELISA) from 0 to 72 h postpartum in 18 of the neonatal foals. The incidence of dystocia and premature placental separation was higher in induced mares (2 of 11 and 1 of 11 versus 0 of 15 and 0 of 15, respectively), whereas retained fetal membranes were more common in spontaneous foalings (2 of 15 versus 0 of 11). When abnormal foals were excluded (to decrease the influence of endogenous serum AA elevations), serum concentrations of AA increased to the same extent over time in foals with induced versus spontaneous parturition; foals with spontaneous parturition had a mean serum AA concentration of 7.8 microg/mL at birth that increased to a maximum of 58.9 microg/mL at 36 h; foals with induced parturition had a mean serum AA concentration of 5.4 microg/mL at birth that increased to a maximum of 41.4 microg/mL at 48 h. Baseline serum AA concentrations were lower in induced foals. We concluded that inducing parturition with low doses of oxytocin in mares at term did not affect (relative to spontaneous parturition) the temporal dynamics of serum AA concentrations in the normal foal in the first 72 h of life. However, the induction procedure may lead to complications during parturition that, if not detected early, could result in the development of an inflammatory response in the neonate.     

Characterization of rabbit Pasteurella multocida isolates by use of whole-cell, outer-membrane, and polymerase chain reaction typing.

PURPOSE: To characterize Pasteurella multocida isolates from laboratory rabbits using serotyping, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of whole-cell proteins (WCPs) and outer-membrane proteins (OMPs), and polymerase chain reaction (PCR) fingerprinting. METHODS: Fifty isolates were obtained from five sources: ATCC (1), Oklahoma (4), Michigan (9), Minnesota (7), and Texas (29). The PCR fingerprinting was conducted using two minisatellite probes for M13 and a modified M13 core sequence and two microsatellite probes--(GTG)5 and (GACA)4. RESULTS: Forty-five isolates were serogroup A, and five were serogroup D. Ten WCP patterns (W1-W10) with one variation (W1a) and 10 OMP (OM1-OM10) patterns were found. Primers M13 phage, modified M13 phage, (GTG)5, and (GACA)4 generated 7, 9, 5, and 9 fingerprint types, respectively. Combination of WCP, OMP, and PCR fingerprint results yielded 39 groups with a discrimination index of 0.98. The PCR fingerprint results generally indicated clonal association among isolates within geographic locations except for the isolates from Texas, which varied markedly in PCR fingerprint types. CONCLUSION: Single primer PCR fingerprinting provided a simple and rapid means of typing P. multocida isolates from laboratory rabbits. Combinations of conventional and molecular typing enhanced differentiation among P. multocida isolated from rabbits with pasteurellosis.