Research Progress Reports, 1962. Fruit and Vegetable Processing and Technology Division, Department of Horticulture.

Tomato variety evaluation for processing, 1962 / W. A. Gould, J. R. Geisman and Wade Schulte -- Evaluation of snap bean varieties for processing, 1962 / Wilbur A. Gould -- Handling and holding studies of mechanically harvested tomatoes. 1. Processed product quality / W. A. Gould, W. D. Bash, J. R. Geisman, D. E. Yingst, G. A. Marlowe and W. N. Brown -- Handling and holding studies of mechanically harvested tomatoes. 2. Spore counts / Winston D. Bash and W. A. Gould -- Handling and holding studies of mechanically harvested tomatoes. 3. pH / Winston D. Bash and W. A. Gould -- Handling and holding studies of mechanically harvested tomatoes. 4. Chlorine residuals / Donald E. Yingst and W. A. Gould -- Removal of insects and residues from sweet corn by washing techniques / J. R. Geisman and W. A. Gould -- Removal of pesticides and radioactive fallout from fruits and vegetables / J. R. Geisman, R. P. Blackmore, R. W. Hirzel and W. S. Stinson -- The effect of apple variety and browning prevention treatments during preparation on the quality of frozen apple pies / D. Robert Davis and James F. Gallander -- Effect of three tomato peeling methods on efficiency and product quality / Wade A. Schulte and W. A. Gould -- Effects of cooling rates on vacuum of canned pumpkin / Winston D. Bash -- New flavors for sauerkraut / J. R. Geisman and Robert Reyda -- Flavor of tomato juice / Wilbur A. Gould, Natholyn Dalton and John Hal Johnson -- Fruit juice blends offer a promising new field for apple cider / D. Robert Davi

A Single Nucleotide Change Affects Fur-Dependent Regulation of sodB in H. pylori

Helicobacter pylori is a significant human pathogen that has adapted to survive the many stresses found within the gastric environment. Superoxide Dismutase (SodB) is an important factor that helps H. pylori combat oxidative stress. sodB was previously shown to be repressed by the Ferric Uptake Regulator (Fur) in the absence of iron (apo-Fur regulation) [1]. Herein, we show that apo regulation is not fully conserved among all strains of H. pylori. apo-Fur dependent changes in sodB expression are not observed under iron deplete conditions in H. pylori strains G27, HPAG1, or J99. However, Fur regulation of pfr and amiE occurs as expected. Comparative analysis of the Fur coding sequence between G27 and 26695 revealed a single amino acid difference, which was not responsible for the altered sodB regulation. Comparison of the sodB promoters from G27 and 26695 also revealed a single nucleotide difference within the predicted Fur binding site. Alteration of this nucleotide in G27 to that of 26695 restored apo-Fur dependent sodB regulation, indicating that a single base difference is at least partially responsible for the difference in sodB regulation observed among these H. pylori strains. Fur binding studies revealed that alteration of this single nucleotide in G27 increased the affinity of Fur for the sodB promoter. Additionally, the single base change in G27 enabled the sodB promoter to bind to apo-Fur with affinities similar to the 26695 sodB promoter. Taken together these data indicate that this nucleotide residue is important for direct apo-Fur binding to the sodB promoter

Diversity in Functional Organization of Class I and Class II Biotin Protein Ligase

The cell envelope of Mycobacterium tuberculosis (M.tuberculosis) is composed of a variety of lipids including mycolic acids, sulpholipids, lipoarabinomannans, etc., which impart rigidity crucial for its survival and pathogenesis. Acyl CoA carboxylase (ACC) provides malonyl-CoA and methylmalonyl-CoA, committed precursors for fatty acid and essential for mycolic acid synthesis respectively. Biotin Protein Ligase (BPL/BirA) activates apo-biotin carboxyl carrier protein (BCCP) by biotinylating it to an active holo-BCCP. A minimal peptide (Schatz), an efficient substrate for Escherichia coli BirA, failed to serve as substrate for M. tuberculosis Biotin Protein Ligase (MtBPL). MtBPL specifically biotinylates homologous BCCP domain, MtBCCP87, but not EcBCCP87. This is a unique feature of MtBPL as EcBirA lacks such a stringent substrate specificity. This feature is also reflected in the lack of self/promiscuous biotinylation by MtBPL. The N-terminus/HTH domain of EcBirA has the self-biotinable lysine residue that is inhibited in the presence of Schatz peptide, a peptide designed to act as a universal acceptor for EcBirA. This suggests that when biotin is limiting, EcBirA preferentially catalyzes, biotinylation of BCCP over self-biotinylation. R118G mutant of EcBirA showed enhanced self and promiscuous biotinylation but its homologue, R69A MtBPL did not exhibit these properties. The catalytic domain of MtBPL was characterized further by limited proteolysis. Holo-MtBPL is protected from proteolysis by biotinyl-5′ AMP, an intermediate of MtBPL catalyzed reaction. In contrast, apo-MtBPL is completely digested by trypsin within 20 min of co-incubation. Substrate selectivity and inability to promote self biotinylation are exquisite features of MtBPL and are a consequence of the unique molecular mechanism of an enzyme adapted for the high turnover of fatty acid biosynthesis

Global parasite trafficking:Asian Gyrodactylus (Monogenea) arrived to the U.S.A. via invasive fish Misgurnus anguillicaudatus as a threat to amphibians

Abstract A monogenean flatworm Gyrodactylus jennyae Paetow, Cone, Huyse, McLaughlin & Marcogliese, 2009 was previously described as a pathogen on bullfrog Lithobates catesbeianus Shaw, 1802, in a Canadian captive population originating in Missouri, U.S.A. The ITS barcoding of G. jennyae showed relatedness to Asian Gyrodactylus macracanthus Hukuda 1940, a parasite of the Asian loach Misgurnus anguillicaudatus Cantor, 1842. The resulting suggestion that the globally invasive pet-trade of fish may be a mechanism for arrival of Gyrodactylus species to North America provided the framework for the current study. The present study was undertaken following the discovery of two other species of Gyrodactylus in a population of illegally introduced M. anguillicaudatus in New York State. Here the invasion hypothesis was tested via DNA sequencing of the ITS of the two Gyrodactylus species obtained from M. anguillicaudatus from New York, termed Gyrodactylus sp. A and Gyrodactylus sp. B. Both Gyrodactylus sp. A and Gyrodactylus sp. B were closely related to G. jennyae and G. macracanthus, and all belong to a molecularly well-supported monophyletic Asian freshwater group. In conclusion, this invasive fish has trafficked at least three parasite species to the U.S.A., one of them also found on frog. This route from the Asian wetlands to other continents is similar to that of amphibian chytrid fungi of genus Batrachochytrium Longcore, Pessier & Nichols, 1999

EPCR-PAR1 biased signaling regulates perfusion recovery and neovascularization in peripheral ischemia

Blood clot formation initiates ischemic events, but coagulation roles during postischemic tissue repair are poorly understood. The endothelial protein C receptor (EPCR) regulates coagulation, as well as immune and vascular signaling, by protease activated receptors (PARs). Here, we show that endothelial EPCR-PAR1 signaling supports reperfusion and neovascularization in hindlimb ischemia in mice. Whereas deletion of PAR2 or PAR4 did not impair angiogenesis, EPCR and PAR1 deficiency or PAR1 resistance to cleavage by activated protein C caused markedly reduced postischemic reperfusion in vivo and angiogenesis in vitro. These findings were corroborated by biased PAR1 agonism in isolated primary endothelial cells. Loss of EPCR-PAR1 signaling upregulated hemoglobin expression and reduced endothelial nitric oxide (NO) bioavailability. Defective angiogenic sprouting was rescued by the NO donor DETA-NO, whereas NO scavenging increased hemoglobin and mesenchymal marker expression in human and mouse endothelial cells. Vascular specimens from patients with ischemic peripheral artery disease exhibited increased hemoglobin expression, and soluble EPCR and NO levels were reduced in plasma. Our data implicate endothelial EPCR-PAR1 signaling in the hypoxic response of endothelial cells and identify suppression of hemoglobin expression as an unexpected link between coagulation signaling, preservation of endothelial cell NO bioavailability, support of neovascularization, and prevention of fibrosis

Research Progress Reports, 1963, Fruit and Vegetable Processing and Technology Division, Department of Horticulture.

Evaluation of tomato varieties for processing, 1963 / W. A. Gould, J. R. Geisman, Winston Bash, Donald Yingst, and W. N. Brown -- Evaluation of snap bean varieties for processing, 1963 / Wilbur A. Gould -- Handling and holding studies of mechanically harvested tomatoes / W. A. Gould, Winston Bash, Donald Yingst, J. R. Geisman and W. N. Brown -- Processing and varietal effects on frozen apple pies / J. F. Gallander and H. L. Stammer -- Removal of pesticide residues from fruits and vegetables / J. R. Geisman and R. P. Blackmore -- Effect of various peeling methods on the removal of simulated radioactive fallout from potatoes / W. S. Stinson and J. R. Geisman -- Effect of detergents on the growth and thermal resistance of Bacillus thermoacidurans / Esmat El Ashwah and W. A. Gould -- Effect of potassium sorbate and vitamin K-5 on apple juice inoculated with yeasts of Saccharomyces genus / Almed Shahine and W. A. Gould -- New sauerkraut products / R. Reyda and J. R. Geisman -- Cider-strawberry fruit juice blend / J. F. Gallander and H. L. Stammer -- The isolation and identification of some volatile constituents of tomato juice / John Hal Johnson and W. A. Goul