First report of Alternaria sp. PMK1: causative agent of leaf spot disease on parthenium weed

Congress grass, Parthenium hysterophorus L. (Asteraceae, tribe Heliantheae), is an erect and much branched annual or ephemeral herb, known as a pest for environmental, medical and agricultural reasons. Survey of fungal pathogens of P. hysterophorus was conducted in Kurukshetra and its adjoining areas, Haryana, India. A leaf spot pathogen Alternaria sp. PMK1 was isolated. Koch's postulates were performed and proved that this isolate was pathogenic to this weed. This is the first report of this pathogen causing leaf spot on parthenium weed.     

In Planta Single-Molecule Pull-Down Reveals Tetrameric Stoichiometry of HD-ZIPIII:LITTLE ZIPPER Complexes

Deciphering complex biological processes markedly benefits from approaches that directly assess the underlying biomolecular interactions. Most commonly used approaches to monitor protein-protein interactions typically provide nonquantitative readouts that lack statistical power and do not yield information on the heterogeneity or stoichiometry of protein complexes. Single-molecule pull-down (SiMPull) uses single-molecule fluorescence detection to mitigate these disadvantages and can quantitatively interrogate interactions between proteins and other compounds, such as nucleic acids, small molecule ligands, and lipids. Here, we establish SiMPull in plants using the HOMEODOMAIN LEUCINE ZIPPER III (HD-ZIPIII) and LITTLE ZIPPER (ZPR) interaction as proof-of-principle. Colocalization analysis of fluorophore-tagged HD-ZIPIII and ZPR proteins provides strong statistical evidence of complex formation. In addition, we use SiMPull to directly quantify YFP and mCherry maturation probabilities, showing these differ substantially from values obtained in mammalian systems. Leveraging these probabilities, in conjunction with fluorophore photobleaching assays on over 2000 individual complexes, we determined HD-ZIPIII:ZPR stoichiometry. Intriguingly, these complexes appear as heterotetramers, comprising two HD-ZIPIII and two ZPR molecules, rather than heterodimers as described in the current model. This surprising result raises new questions about the regulation of these key developmental factors and is illustrative of the unique contribution SiMPull is poised to make to in planta protein interaction studies.     

A Minisatellite Sequence within the Propeptide Region of the Vacuolar Carboxypeptidase Y Gene of Schizosaccharomyces pombe

We describe the presence of a minisatellite sequence that displays length polymorphisms in the fission yeast Schizosaccharomyces pombe. The minisatellite sequence was found to reside within the propeptide region of the vacuolar carboxypeptidase Y gene. The minisatellite sequence, which was found only at a single locus, was mitotically stable and displayed length polymorphisms between the two varieties of S. pombe (S. pombe var. pombe and S. pombe var. malidevorans). The minisatellite sequence, however, appeared to be species specific and was absent in other members of the Schizosaccharomyces genus. This report constitutes the first experimental demonstration of the presence of such sequences in yeasts.     

Selecting a more realistic uncertainty factor: Reducing compounding effects of multiple uncertainties

Available toxicology datasets provide a unique opportunity to validate some of the currently used Uncertainty Factors in the development of acceptable exposure levels for noncancer effects. Toxicity studies from two separate sources, the FAO/WHO database on pesticides (1978–1987) and the Monsanto database (through 1988) were chosen to evaluate three of the five currently used Uncertainty Factors. Interspecies differences in NOELs between the three mammalian species evaluated are equal to or less than a factor of 10 for both the FAO/WHO data and the Monsanto data in greater than 90% of the cases evaluated. Median values for the comparison of interspecies NOELs were 3.0 or less for all comparisons except the comparison between the mouse and rat for the Monsanto dataset where the median value was 7.5. Analyses of the Monsanto toxicity database show that the reprotoxicity NOELs were always equivalent to or higher than the chronic or subchronic NOELs for the same material. Therefore, even without conduct of a specific study to address reproductive effects, reasonable protection from adverse reproductive effects can be afforded by use of either subchronic or chronic study NOELs without application of UFD. The median ratio of subchronic NOELs and chronic NOELs was 4, and for a majority of the studies the difference between the NOELs was within one order of magnitude. Our analysis aids in validating the assumption that the upperbound for individual uncertainties maybe accounted for by use of 10‐fold uncertainty factors. However, the current U.S. Environmental Protection Agency (USEPA) reference doses/concentrations may be overly conservative because upperbounds of each of the uncertainty factors are used and each of the uncertainty factors are considered to be independent variables. Because uncertainties are probably not independent variables, the influence of compounding upperbounds when multiple uncertainty factors are used is generally only considered when four or more areas of uncertainty are outstanding. When multiple uncertainties exist, we recommend upperbound estimates only be used for the first two Uncertainty Factors, and median values be used to account for the remaining uncertainties.     

Effect of visible light on the mitochondrial inner membrane.

Exposure of mitochondria to visible light in the presence of riboflavin resulted in the initial release of respiratory control, followed by inhibition of electron transport and dissolution of structural integrity. Under these conditions, however, cytochrome c oxidase activity remained unchanged. ATPase activity was stimulated initially and remained in this activitated state even under continued illumination. In submitochondrial preparations, both electron transport and ATPase declined as a function of illumination time; cytochrome c oxidase was not sensitive to light. Enzyme inactivation also occurred to a lesser extent in the absence of riboflavin.     

Proligodendroblast Antigen (POA), a Developmental Antigen Expressed by A007/O4-Positive Oligodendrocyte Progenitors Prior to the Appearance of Sulfatide and Galactocerebroside

Evidence is presented for the immunological identification of a developmental antigen appearing at a critical point in the oligodendroglial lineage. Specifically, monoclonal antibody A007 recognizes cells in the oligodendrocyte lineage at two distinct stages. Analyses of purified lipid standards and lipid extracts from galactocerebroside-positive (GalC+) oligodendrocytes by enzyme-linked immunosorbent assay, lipid dot blot, and immuno-TLC demonstrated that A007 recognizes sulfatide (SUL) and seminolipid. However, neither 35SO4 incorporation into SUL nor SUL accumulation could be detected in A007-positive cells lacking galactocerebroside (i.e., A007+GalC- progenitor cells) present early in development. These data suggest that A007 also recognizes an antigen, named proligodendroblast antigen (POA), that appears during the late stage of oligodendrocyte progenitor development prior to the expression by oligodendrocytes of SUL and GalC. We have previously reported that monoclonal antibody O4 also recognizes not only SUL and seminolipid, but in addition an antigen that appears prior to the expression of SUL and galactocerebroside. In the present study all A007+ cells were also O4+ (and vice versa), and the developmental patterns of the two antibodies appeared to be identical. We conclude that (1) A007 is similar or identical to O4 with respect to its antigenic specificity, and (2) during oligodendrocyte lineage progression both antibodies react first with antigen POA on the surface of the oligodendrocyte progenitor cell prior to the expression of SUL [i.e., A007+O4+(POA+)SUL-GalC- proligodendroblasts], and only later with SUL as terminally differentiating oligodendrocytes emerge (i.e., A007+O4+SUL+GalC+ oligodendrocytes).     

The quantitation of rabies-specific antibodies. I. Modified counter immunoelectrophoresis. A rapid and sensitive method

Modified counter immunoelectrophoresis was standardized with respect to dilution of tissue culture antigen and indicator serum, the incubation time for neutralization and the effect of an electric current. The technique was found to be sensitive enough to detect a minimum level of antibodies (0.5 IU/ml) by using a 16 mA current per slide for 2 h, indicator serum of 15 IU/ml and the use of an antigen at a concentration of 1:35. Above all, the incubation period did not affect the neutralization of the virus. The test was also applied to the detection of rabies-specific antibody levels in 73 human sera. The test was found to be simple, quick and economical for titration of rabies antibodies.     

dbGAPs: A comprehensive database of genes and genetic markers associated with psoriasis and its subtypes

Psoriasis is a systemic hyperproliferative inflammatory skin disorder, although rarely fatal but significantly reduces quality of life. Understanding the full genetic component of the disease association may provide insight into biological pathways as well as targets and biomarkers for diagnosis, prognosis and therapy. Studies related to psoriasis associated genes and genetic markers are scattered and not easily amendable to data-mining. To alleviate difficulties, we have developed dbGAPs an integrated knowledgebase representing a gateway to psoriasis associated genomic data. The database contains annotation for 202 manually curated genes associated with psoriasis and its subtypes with cross-references. Functional enrichment of these genes, in context of Gene Ontology and pathways, provide insight into their important role in psoriasis etiology and pathogenesis. The dbGAPs interface is enriched with an interactive search engine for data retrieval along with unique customized tools for Single Nucleotide Polymorphism (SNP)/indel detection and SNP/indel annotations. dbGAPs is accessible at http://www.bmicnip.in/dbgaps/.