Nucleoside diphosphate kinase of Trypanosoma brucei

Nucleoside diphosphate kinase (NDPK) is a highly conserved, multifunctional enzyme. Its originally described function is the phosphorylation of nucleoside diphosphates to the corresponding triphosphates, using ATP as the phosphate donor and a high-energy phosphorylated histidine residue as the reaction intermediate. More recently, a host of additional functions of NDPK have been discovered. Some of these correlate with the capacity of NDPK to transphosphorylate other proteins, in a manner reminiscent of bacterial two-component systems. Other functions may be mediated by direct DNA-binding of NDPK.This study describes the identification of NDPK from the parasitic protozoon Trypanosoma brucei. The genome of this major disease agent contains a single gene for NDPK. The predicted amino acid sequence of the trypanosomal enzyme is highly conserved with respect to all other species. The protein is constitutively expressed and is present in procyclic and in bloodstream forms. Immunofluorescence and immuno-electron microscopy demonstrate that trypanosomal NDPK (TbNDPK) is predominantly localized in the cell nucleus. Histidine phosphorylation of TbNDPK is essentially resistant to the experimental compound LY266500, a potent inhibitor of histidine phosphorylation of trypanosomal succinyl coenzyme A synthase.     

Mixed Infections of Bacillus subtilis Involving Bacteriophages SP82 and β22

Progeny yields and the synthesis of nucleic acids have been investigated in two strains of Bacillus subtilis mixedly infected with two unrelated phages, SP82 and beta22. When B. subtilis strain 168 was the host, the first phage added dominated the infection; when B. subtilis strain SB11 was the host, beta22 produced progeny even when added to cells 5 min after infection with SP82. Dominance in these mixed infections could be correlated with qualitative and quantitative differences in the synthesis of phage-specific RNAs.     

Dominance Relationships in Mixedly Infected Bacillus subtilis

The progeny released from Bacillus subtilis cells mixedly infected with bacteriophages beta22, SP82, and SP02(c1) have been studied at varying multiplicities of infection and orders of addition and with different host strains of the bacterium. In B. subtilis 168, SP02(c1) was subordinate to both SP82 and beta22 and did not yield significant numbers of progeny even when added 5 min before the superior phage. Dominance in mixed infections of beta22 and SP82 was host-dependent. In B. subtilis 168, SP82 was dominant and greatly reduced the yield of beta22 if added simultaneously or before the subordinate partner. However, in the same mixed infection in B. subtilis SB11, beta22 was the dominant phage and totally suppressed the production of SP82 even when added 5 min after the latter.     

Competition between two stream dwelling filter-feeders,Hydropsyche oslari and Simulium virgatum

Summary This study investigated whether interspecific competition affected the abundance and distribution of Hydropsyche oslari and Simulium virgatum. In addition, I studied the mechanisms of competition between the two taxa. H. oslari was the superior competitor when compared to S. virgatum because the presence of H. oslari in boulder habitats in Refugio Creek altered the microdistribution and depressed numbers of S. virgatum. Hydropsyche preempted space and was aggressive towards Simulium. Simulium avoided both hydropsychid larvae and their nets. This avoidance behaviour was reinforced by aggression from H. oslari. Possible immediate reasons why simuliids avoided Hydropsyche nets included 1) Hydropsyche attacked Simulium, 2) occupied nets interfered with feeding through increased turbulence or lower renewal rate for food, and 3) Simulium preferred to settle with conspecifics.     

Effect of Zytron and Its Degradation Products on Soil Microorganisms

There was no adverse effect of Zytron, o-2,4-dichlorophenyl o-methyl isopropyl phosphoramidothioate, a herbicide, upon molds, actinomycetes, and soil bacteria in field plots, or upon selected soil microorganisms in model systems. 2,4-Dichlorophenol, a degradation product, was found to be toxic at levels above 10 ppm to molds, but levels this high were not found in soil from treated plots. Aspergillus clavatus degraded both Zytron and 2,4-dichlorophenol. Sodium o-methyl isopropyl phosphoramidothioate, another degradation product of Zytron, stimulated the growth of a species of Penicillium.     

Amino-caprolactam γ-secretase inhibitors showing potential for the treatment of Alzheimer's disease

Herein we describe the structure-activity relationship (SAR) of amino-caprolactam analogs derived from amino-caprolactam benzene sulfonamide 1, highlighting affects on the potency of γ-secretase inhibition, selectivity for the inhibition of APP versus Notch processing by γ-secretase and selected pharmakokinetic properties. Amino-caprolactams that are efficacious in reducing the cortical Aβ_x-40 levels in FVB mice via a single 100 mpk IP dose are highlighted.     

MAHRP2, an exported protein of Plasmodium falciparum,is an essential component of Maurer's cleft tethers

Upon invasion into erythrocytes, the malaria parasite Plasmodium falciparum must refurbish the host cell. The objective of this study was to elucidate the location and function of MAHRP2 in these processes. Using immunofluorescence and immunoelectron microscopy we showed that the membrane‐associated histidine‐rich protein‐2 (MAHRP2) is exported during this process to novel cylindrical structures in the erythrocyte cytoplasm. We hypothesize that these structures tether organelles known as Maurer's clefts to the erythrocyte skeleton. Live cell imaging of parasite transfectants expressing MAHRP2–GFP revealed both mobile and fixed populations of the tether‐like structures. Differential centrifugation allowed the enrichment of these novel structures. MAHRP2 possesses neither a signal peptide nor a PEXEL motif, and sequences required for export were determined using transfectants expressing truncated MAHRP2 fragments. The first 15 amino acids and the histidine‐rich N‐terminal region are necessary for correct trafficking of MAHRP2 together with a predicted hydrophobic region. Solubilization studies showed that MAHRP2 is membrane associated but not membrane spanning. Several attempts to delete the mahrp2 gene failed, indicating that the protein is essential for parasite survival.