Potential for biological control of Heterodera schachtii by Pochonia chlamydosporia var. chlamydosporia on sugar beet

The biological control potential of an isolate of Pochonia chlamydosporia var. chlamydosporia against Heterodera schachtii was examined by assessing the percentage of females and cysts that became infected on water agar, the effect of culture filtrate on juvenile mobility, and the effects of the fungus on the final population of the nematode on sugar beet under greenhouse conditions. After 3 weeks at 20°C, 74 and 95% of the eggs within cysts and females, respectively, were colonised by the fungus on water agar. The full concentration of the fungal filtrate from cultures in malt extract broth killed only 12% of the juveniles after 24 h at 25°C. In the greenhouse experiment, adding 16,000 chlamydospores of the fungus per gram of soil as either colonised barley grains or spores reduced the final number of females on roots of sugar beet by 50 and 66%, respectively, after 3 months. The reproduction factor was reduced to ×2 in spore-treated soil compared with ×5 in the untreated control, and 18% of the eggs in spore-treated soil were colonised by fungal mycelium. Generally, P. chlamydosporia var. chlamydosporia was more efficient at reducing the nematode population when applied as spores without any substrate than when used as colonised barley grains.     

Clustering of risk factors and social class in childhood and adulthood in British women's heart and health study: cross sectional analysis

Objective To examine co-occurrence and clustering of risk factors used in the Framingham equation by social class in childhood and adult life.Design Cross sectional study.Setting 23 towns across England, Wales, and Scotland.Participants 2936 women aged 60-79 years.Main outcome measures Prevalence of risk factors (hypertension, obesity, smoking, left ventricular hypertrophy on electrocardiography, diabetes, and low concentration of high density cholesterol); ratios of observed to expected frequencies of clusters of risk factors.Results Risk factors were more common in women from manual social classes in either childhood or adult life, and the co-occurrence of three or four of these risk factors was greater among more disadvantaged groups. Within the four socioeconomic groups, these risk factors occurred together more than would be expected from their individual frequency distributions, indicating that they were clustered. The extent of this clustering was similar in all four social class groups.Conclusions Clustering of risk factors included in the Framingham risk function occurs in all social class groups, but the lack of social patterning makes it unlikely that clustering is an explanation of socioeconomic inequalities in cardiovascular disease. As the proportion of women with co-occurrence of risk factors is greatest in those from manual social class in childhood, this measure of socioeconomic position might prove useful in risk prediction.     

A study on the bacterial photo-toxicity of phenothiazinium based photosensitisers

"Comet assay" showed light activated (3.15 Jcm-2 over 30 min) phenothiazinium based photosensitisers (PhBPs) to induce photo-damage of Staphylococcus aureus DNA, as indicated by DNA "tails" between 80 and 120 microm. In general, PhBPs exhibited significant singlet oxygen yields (Phi(DeltaPhBP)>0.7), suggesting the use of type II mechanisms of photo-oxidation. However, the photodynamic action of PhBPs on DNA showed generally insignificant production of 7,8-dihydro-8-oxo-2'-deoxyguanosine, normally a major product of type II DNA photo-oxidation. These combined results show DNA to be a major site of action of PhBPs and suggest that this action may involve type II attack on a nucleoside(s) other than guanosine.     

PKCε inhibits the hyperglycemia-induced apoptosis signal in adult rat ventricular myocytes

Recruitment of the protein kinase C (PKC) family of isozymes is an integral component of the signaling events that direct cardiac phenotype expressed during postnatal development and in response to pathologic stimuli. Hyperglycemia is a potent activating signal for cardiac PKC isozymes and induces the apoptosis program in cardiac muscle cells. To determine whether cardiac PKC isozymes modulate transmission of the hyperglycemia apoptosis signal, we have employed isozyme-specific peptide modulators to selectively inhibit (PKC _I/_II, and ) or activate (PKC). PKC peptides were delivered to primary cultures of serum starved adult rat ventricular myocytes (ARVM), by conjugation to the homeodomain of drosophila antennapedia. As expected, hyperglycemia induced a 35% increase in ARVM apoptosis. Peptide inhibitors of PKC _I/_II and blocked transmission of the hyperglycemia apoptosis signal, whereas the isozyme specific inhibitor of PKC (V1-2) did not alter the magnitude of glucose-induced ARVM apoptosis. Alternatively, the PKC translocation activator (RACK) abolished hyperglycemia-induced apoptosis, strongly suggesting a cardioprotective role for PKC in this system. Therefore, we conclude that cardiac PKC isozymes modulate hyperglycemia-induced apoptosis and activation of cardiac PKC protects ARVM from the hyperglycemia-induced death signal. (Mol Cell Biochem 268: 169–173, 2005)     

Residue 219 impacts on the dynamics of the C-terminal region in glutathione transferase A1-1: implications for stability and catalytic and ligandin functions

The C-terminal region in class alpha glutathione transferases (GSTs) modulates the catalytic and nonsubstrate ligand binding functions of these enzymes. Except for mouse GST A1-1 (mGST A1-1), the structures of class alpha GSTs have a bulky aliphatic side chain topologically equivalent to Ile219 in human GST A1-1 (hGST A1-1). In mGST A1-1, the corresponding residue is an alanine. To investigate the role of Ile219 in determining the conformational dynamics of the C-terminal region in hGST A1-1, the residue was replaced by alanine. The substitution had no effect on the global structure of hGST A1-1 but did reduce the conformational stability of the C-terminal region of the protein. This region could be stabilized by ligands bound at the active site. The catalytic behavior of hGST A1-1 was significantly compromised by the I219A mutation as demonstrated by reduced enzyme activity, increased K(m) for the substrates glutathione (GSH) and 1-chloro-2,4-dinitrobenzene (CDNB), and reduced catalytic efficiencies. Inhibition studies also indicated that the binding affinities for product and substrate analogues were dramatically decreased. The affinity of the mutant for GSH was, however, only slightly increased, indicating that the G-site was unaltered by the mutation. The binding affinity and stoichiometry for the anionic dye 8-anilino-1-naphthalene sulfonate (ANS) was also not significantly affected by the I219A mutation. However, the lower DeltaC(p) for ANS binding to the mutant (-0.34 kJ/mol per K compared with -0.84 kJ/mol per K for the wild-type protein) suggests that ANS binding to the mutant results in the burial of less hydrophobic surface area. Fluorescence data also indicates that ANS bound to the mutant is more prone to quenching by water. Overall, the data from this study, together with the structural details of the C-terminal region in mGST A1-1, show that Ile219 is an important structural determinant of the stability and dynamics of the C-terminal region of hGST A1-1.     

Signaling role of intracellular iron in NF-kappaB activation

Iron chelators inhibit endotoxin-induced NF-kappaB activation in hepatic macrophages (HMs), suggesting a role for the intracellular chelatable pool of iron in NF-kappaB activation. The present study tested this hypothesis. Analysis of Fe(59)-loaded HMs stimulated with lipopolysaccharide (LPS), revealed a previously unreported, transient rise in intracellular low molecular weight (LMW).Fe(59) complex ([LMW.Fe](i)) at /=15 min) and NF-kappaB (>/=30 min) activation. Iron chelators (1,2-dimethyl-3-hydroxypyridin-4-one and N,N'-bis-2-hydroxybenzylethylenediamine-N,N'-diacetic acid) abrogated the [LMW.Fe](i) response and IKK and NF-kappaB activation. The [LMW.Fe](i) response was also observed in tumor necrosis factor alpha (TNFalpha)-stimulated HMs and RAW264.7 cells treated with LPS and interferon-gamma but not in primary rat hepatocytes or myofibroblastic cells exposed to LPS or TNFalpha. Both [LMW.Fe](i) response and IKK activation in LPS-stimulated HMs were inhibited by diphenylene iodonium (nonspecific inhibitor for flavin-containing oxidases), l-N(6)-(1-iminoethyl)lysine (selective iNOS inhibitor), and adenoviral-mediated expression of a dominant negative mutant of Rac1 or Cu,Zn-superoxide dismutase, suggesting the role of (.)NO and O(2)() in mediating the iron signaling. In fact, this inhibition was recapitulated by a cell-permeable scavenger of ONOO(-), 5,10,15,20-tetrakis (4-sulfonatophenyl)porphyrinato iron (III) chloride. Conversely, ONOO(-) alone induced both [LMW.Fe](i) response and IKK activation. Finally, direct addition of ferrous iron to cultured HMs activated IKK and NF-kappaB. These results support a novel signaling role for [LMW.Fe](i) in IKK activation, which appears to be induced by ONOO(-) and selectively operative in macrophages.     

Two ellagitannins from Punica granatum heartwood

In the course of studies on polyphenol metabolism in Punica granatum heartwood two new ellagitannins, diellagic acid rhamnosyl (1-->4) glucopyranoside and 5-O-galloylpunicacortein D were isolated and characterized together with four known tannin metabolites, punicacortein D, punicalin, punicalagin and 2-O-galloylpunicalin. Structures of the isolated compounds were established by chromatography, chemical degradation, UV and 1D/ 2D 1H/13C NMR spectroscopy.     

Effects of shoot and root application of thiamin on salt-stressed sunflower plants

Plants of sunflower (Helianthus annuus L. cv Giza2) were salt-stressed with a combination of NaCl and CaCl₂ inconcentrations having different osmotic potentials (s from 0 to –1.0MPa) and were treated with 5 and 10mg L^–1 of thiamin either sprayed on the shoot orapplied to the root. The membranes of leaf discs from salt-stressed plantsappeared to be less stable (more injured) under heat(51°C) and drought (40% polyethylene glycol6000) stresses than control plants. Salinity slowed the rate of growth (lengthand dry mass production), lowered leaf relative water content (RWC) and leafandroot water potential (w), decreased the contents of chlorophyll (Chl),soluble sugars (SS) and the K⁺/Na⁺ ratio butenhanced total free amino acids (TAA), Na⁺,Ca²⁺and Cl^– accumulation in the shoot and root system. Root orshoot application of thiamin reduced membrane injury by either heat ordehydration stress, lowered leaf w, improved uptake of K⁺,and increased leaf RWC, Chl, SS, TAA contents and dry mass production. Theeffects of salinity (s), thiamin (Thi.) and their interaction(s×Thi) on the parameters tested were significant.Salinity was dominant (as indicated by ² values) in affectingthe contents of Ca²⁺, Cl^–, TAA and membranestability to heat and leaf w. The role of thiamin was dominant forNa⁺, K⁺ and SS contents and the contribution ofinteraction was dominant for growth parameters, Chl. and root w.     

Characterization of zebrafish merlot/chablis as non-mammalian vertebrate models for severe congenital anemia due to protein 4.1 deficiency

The red blood cell membrane skeleton is an elaborate and organized network of structural proteins that interacts with the lipid bilayer and transmembrane proteins to maintain red blood cell morphology, membrane deformability and mechanical stability. A crucial component of red blood cell membrane skeleton is the erythroid specific protein 4.1R, which anchors the spectrin-actin based cytoskeleton to the plasma membrane. Qualitative and quantitative defects in protein 4.1R result in congenital red cell membrane disorders characterized by reduced cellular deformability and abnormal cell morphology. The zebrafish mutants merlot (mot) and chablis (cha) exhibit severe hemolytic anemia characterized by abnormal cell morphology and increased osmotic fragility. The phenotypic analysis of merlot indicates severe hemolysis of mutant red blood cells, consistent with the observed cardiomegaly, splenomegaly, elevated bilirubin levels and erythroid hyperplasia in the kidneys. The result of electron microscopic analysis demonstrates that mot red blood cells have membrane abnormalities and exhibit a severe loss of cortical membrane organization. Using positional cloning techniques and a candidate gene approach, we demonstrate that merlot and chablis are allelic and encode the zebrafish erythroid specific protein 4.1R. We show that mutant cDNAs from both alleles harbor nonsense point mutations, resulting in premature stop codons. This work presents merlot/chablis as the first characterized non-mammalian vertebrate models of hereditary anemia due to a defect in protein 4.1R integrity.