Biosynthesis of Depsipeptide Mycotoxins in Fusarium

The cyclic hexadepsipeptide enniatin is known as a phytopathogenic compound from Fusaria causing necrosis and wilt. The molecule consists of three alternating residues each of a branched chain amino acid and D-hydroxyisovaleric acid (D-Hiv). Enniatins are synthesized by a 347kDa multienzyme (enniatin synthetase) via a thiol template mechanism. The corresponding gene esyn1 has an open reading frame of 9393 nucleotides and harbours two modules, one responsible for D-hydroxy acid activation and one for L-amino acid activation with an integrated N-methyltransferase domain. Such methyltransferases build an homologous group among N-methyl peptide synthetases. Enniatins are synthesized by step-wise condensation of dipeptidol building blocks in an iterative manner resembling fatty acid synthesis. A key enzyme in enniatin biosynthesis is the NADPH-dependent D-2-hydroxyisovalerate dehydrogenase, that supplies enniatin synthetase with D-Hiv. Enniatins contribute to the wilt toxic character of Fusaria. Virulence was significantly reduced in F. avenaceum after disruption of the esyn1 gene.     

Plasma atrial natriuretic peptide concentration in warmblood horses with heart valve regurgitations

ObjectivesThis study measured plasma atrial natriuretic peptide (ANP) concentration in horses with heart valve regurgitations (HVR) with and without atrial and ventricular dilatation.BackgroundIn humans and small animals, plasma ANP concentration is increased in heart disease and correlates with the severity of clinical signs and heart enlargement.Animals, materials and methodsTen healthy horses (control) and 36 horses with HVR were evaluated by auscultation, electrocardiography, echocardiography, and determination of plasma ANP.ResultsControl horses demonstrated mean plasma ANP concentration of 21 ± 5.4 pg/mL. Of the 36 horses with HVR, 17 horses possessed normal echocardiographic heart size (group 1), 10 horses had a left atrial dilatation (group 2) and 9 horses had both left atrial and ventricular dilatation (group 3). Mean plasma ANP concentration of groups 1, 2 and 3 was 20.1 ± 5.6 pg/mL, 22.9 ± 11.0 pg/mL and 27.6 ± 17.4 pg/mL, respectively. The plasma ANP concentrations of HVR and control horses were not significantly different. The highest ANP concentrations were observed in horses with atrial and ventricular dilatation. No correlation between left atrial or ventricular size, weight, or sex and the plasma ANP concentration was found.ConclusionsNo significant differences in plasma ANP concentration was observed between groups. Further study, especially in horses with clinical signs of heart failure is needed.     

A microsatellite sequence closely linked to the Waxy gene of Oryza sativa

Summary A polymorphic microsatellite locus has been located closely linked to the Waxy gene of rice. Primers were designed to allow detection of the microsatellite by utilising the polymerase chain reaction. In screen of 13 commercial rice varieties, four different alleles were found, demonstrating the potential of this marker in commercial rice breeding for starch quality.Abbreviations RFLP Restriction Fragment Length Polymorphism - PCR Polymerase Chain Reaxtion     

The Share of Southeastern Black Counties in the Southern Rural Renaissance

Some authors have suggested that a leading problem associated with US factory employment for blacks may be that the nonfarm jobs--specifically manufacturing--have tended to go to heavily white counties rather than to the mainly black counties of the Deep South. This study is limited to the 1959-1977 period, the time of the most vigorous nonmetropolitan employment growth. To test the hypothesis that factory jobs have bypassed heavily black counties, a 100% sample of nonmetropolitan counties was used from 13 Southern states for the 2 time periods, 1959-1969 and 1969-1977. Results show that the minority counties were not bypassed by Southern rural factory job growth either in the 1960s or the 1970s, but their gains were not as sizeable as those of more heavily white counties. Both the neoclassical economics of discrimination--with its individualistic "taste for discrimination"--and the stinging critique by Marshall in 1974 that such analysis ignores the important role of institutionalized racism--can "explain the empirical conclusions of this article." Traditional "smokestack-chasing" methods are by no means hopeless. While co-ops and community development corporation strategies can be combined with traditional methods, it is important that foundations, government agencies, anti-poverty programs, and black leadership also focus on projects to competitively attract jobs in the traditional fashion.     

TILLING and Ecotilling the Plant Kingdom

The modem crop scientist has a large amount of available nucleotide sequence information to identify genes of potential agronomic importance. Using reverse genetic approaches, specific genes can be disrupted, and hypotheses regarding gene function directly tested in vivo. Although a number of reverse genetic methods have been introduced, many are limited in application because they are organism-specific, expensive, transgenic, or only transiently disrupt gene function. However, traditional mutagenesis using chemical mutagens has been widely used as a forward genetics strategy to create many new crop plant varieties at relatively low cost. Mutagens such as ethyl methanesulphonate （EMS） cause stable point mutations and thus produce an allelic series of truncation and missense changes that can provide a range of phenotypes （Greene et al., 2003）. TILLING （targeting induced local lesions IN genomes） is a high-throughput reverse genetic strategy that combines traditional mutagenesis and SNP discovery methods （Colbert et al., 2001; McCallum et al., 2000）. To identify mutations, target regions of-l.5 kb are amplified with fluorescently labeled gene specific primers. Heteroduplexes are then formed between wild-type and mutant strands, mismatches are cleaved by incubation with a single-strand specific nuclease,     

Einfluß von Mg- und Ca-Mangel auf Mg- und Ca-Gehalte,Chlorophyllgehalt und Chlorophyllfluoreszenz von Fichtennadeln sowie auf die Mikromorphologie und Benetzbarkeit ihrer epicuticulären Wachse

The influence of Mg- and Ca-deficiency on the micromorphology of epicuticular waxes and on the wettability of spruce needles The influence of Mg- and Ca-deficiency in nutrient solution on epicuticular waxes of needles of Norway spruce (Picea abies [L.] Karst.) was investigated in greenhouse experiments. Spruce clones were cultivated in sand and supplied with nutrient solution. For two vegetation periods, plants of the treatment groups -Mg, -Ca and -Mg/-Ca were provided with only 15 % of the respective mineral element of the control. At the end of the second vegetation period, the nutrient deficiency was documented by reduced Mg- and/or Ca-content of the needles and changes in chlorophyll fluorescence. No effect on micromorphology of epicuticular waxes and wettability of needles of any age was observed following deficient Mg and/or Ca supply.     

Improved sample preparation to determine acrylamide in difficult matrixes such as chocolate powder, cocoa, and coffee by liquid chromatography tandem mass spectroscopy

An improved sample preparation (extraction and cleanup) is presented that enables the quantification of low levels of acrylamide in difficult matrixes, including soluble chocolate powder, cocoa, coffee, and coffee surrogate. Final analysis is done by isotope-dilution liquid chromatography-electrospray ionization tandem mass spectrometry (LC-MS/MS) using d3-acrylamide as internal standard. Sample pretreatment essentially encompasses (a) protein precipitation with Carrez I and II solutions, (b) extraction of the analyte into ethyl acetate, and (c) solid-phase extraction on a Multimode cartridge. The stability of acrylamide in final extracts and in certain commercial foods and beverages is also reported. This approach provided good performance in terms of linearity, accuracy and precision. Full validation was conducted in soluble chocolate powder, achieving a decision limit (CCalpha) and detection capability (CCbeta) of 9.2 and 12.5 microg/kg, respectively. The method was extended to the analysis of acrylamide in various foodstuffs such as mashed potatoes, crisp bread, and butter biscuit and cookies. Furthermore, the accuracy of the method is demonstrated by the results obtained in three inter-laboratory proficiency tests.     

5-HT4(a) receptors avert opioid-induced breathing depression without loss of analgesia

Opiates are widely used analgesics in anesthesiology, but they have serious adverse effects such as depression of breathing. This is caused by direct inhibition of rhythm-generating respiratory neurons in the Pre-Boetzinger complex (PBC) of the brainstem. We report that serotonin 4(a) [5-HT4(a)] receptors are strongly expressed in respiratory PBC neurons and that their selective activation protects spontaneous respiratory activity. Treatment of rats with a 5-HT4 receptor-specific agonist overcame fentanyl-induced respiratory depression and reestablished stable respiratory rhythm without loss of fentanyl's analgesic effect. These findings imply the prospect of a fine-tuned recovery from opioid-induced respiratory depression, through adjustment of intracellular adenosine 3',5'-monophosphate levels through the convergent signaling pathways in neurons.     

Redesigning nature's poisons to create anti-tumor reagents

Immunotoxins are conjugates of cell-reactive antibodies and toxins or their subunits. In this report, the chemistry, biology, pharmacokinetics, and anti-tumor effects of first generation immunotoxins; the preparation of improved second generation immunotoxins that display greater anti-tumor efficacy; and the role of genetic engineering in creating third-generation immunotoxins are discussed.