Molecular Characterization of Protease Activity in Serratia sp. Strain SCBI and Its Importance in Cytotoxicity and Virulence

A newly recognized Serratia species, termed South African Caenorhabditis briggsae isolate (SCBI), is both a mutualist of the nematode Caenorhabditis briggsae KT0001 and a pathogen of lepidopteran insects. Serratia sp. strain SCBI displays high proteolytic activity, and because secreted proteases are known virulence factors for many pathogens, the purpose of this study was to identify genes essential for extracellular protease activity in Serratia sp. strain SCBI and to determine what role proteases play in insect pathogenesis and cytotoxicity. A bank of 2,100 transposon mutants was generated, and six SCBI mutants with defective proteolytic activity were identified. These mutants were also defective in cytotoxicity. The mutants were found defective in genes encoding the following proteins: alkaline metalloprotease secretion protein AprE, a BglB family transcriptional antiterminator, an inosine/xanthosine triphosphatase, GidA, a methyl-accepting chemotaxis protein, and a PIN domain protein. Gene expression analysis on these six mutants showed significant downregulation in mRNA levels of several different types of predicted protease genes. In addition, transcriptome sequencing (RNA-seq) analysis provided insight into how inactivation of AprE, GidA, and a PIN domain protein influences motility and virulence, as well as protease activity. Using quantitative reverse transcription-PCR (qRT-PCR) to further characterize expression of predicted protease genes in wild-type Serratia sp. SCBI, the highest mRNA levels for the alkaline metalloprotease genes (termed prtA1 to prtA4) occurred following the death of an insect host, while two serine protease and two metalloprotease genes had their highest mRNA levels during active infection. Overall, these results indicate that proteolytic activity is essential for cytotoxicity in Serratia sp. SCBI and that its regulation appears to be highly complex

Influence of Temperature on the Physiology and Virulence of the Insect Pathogen Serratia sp. Strain SCBI

The physiology of a newly recognized Serratia species, termed South African Caenorhabditis briggsae Isolate (SCBI), which is both a nematode mutualist and an insect pathogen, was investigated and compared to that of Serratia marcescens Db11, a broad-host-range pathogen. The two Serratia strains had comparable levels of virulence for Manduca sexta and similar cytotoxic activity patterns, but motility and lipase and hemolytic activities differed significantly between them

Inhibition of Escherichia coli chemotaxis by omega-conotoxin, a calcium ion channel blocker

Escherichia coli chemotaxis was inhibited by omega-conotoxin, a calcium ion channel blocker. With Tris-EDTA-permeabilized cells, nanomolar levels of omega-conotoxin inhibited chemotaxis without loss of motility. Cells treated with omega-conotoxin swam with a smooth bias, i.e., tumbling was inhibited

Benchmark of a modified Iterated Perturbation Theory approach on the 3d FCC lattice at strong coupling

The Dynamical Mean-Field theory (DMFT) approach to the Hubbard model requires a method to solve the problem of a quantum impurity in a bath of non-interacting electrons. Iterated Perturbation Theory (IPT) has proven its effectiveness as a solver in many cases of interest. Based on general principles and on comparisons with an essentially exact Continuous-Time Quantum Monte Carlo (CTQMC) solver, here we show that the standard implementation of IPT fails away from half-filling when the interaction strength is much larger than the bandwidth. We propose a slight modification to the IPT algorithm that replaces one of the equations by the requirement that double occupancy calculated with IPT gives the correct value. We call this method IPT-$D$. We recover the Fermi liquid ground state away from half-filling. The Fermi liquid parameters, density of states, chemical potential, energy and specific heat on the FCC lattice are calculated with both IPT-$D$ and CTQMC as benchmark examples. We also calculated the resistivity and the optical conductivity within IPT-$D$. Particle-hole asymmetry persists even at coupling twice the bandwidth. Several algorithms that speed up the calculations are described in appendices.Comment: 17 pages, 15 figures, minor changes to improve clarit

Abundance of Coccinellids and Their Potential Prey in Field-Crop and Grass Habitats in Eastern South Dakota

A rich fauna of coccinellids occurs in eastern South Dakota, but the abundance of some species has declined in association with the establishment of an exotic lady beetle, Coccinella septempunctata (L.) (Coleoptera: Coccinellidae), in the mid-1980s. In this study, coccinellids were sampled within field-crop and grass habitats in eastern South Dakota from 1990 to 1995 to survey for various coccinellid species and to determine any effects of habitat management on abundance. Field crops (maize, wheat-alfalfa intercrop, and alfalfa) were subjected to high, intermediate, or low crop-management intensity (CMI), and grass habitats were managed for stands of warm season, cool season, or mixed species. A total of 1,306 adult and 155 larval coccinellids were collected. Four native species (Coleomegilla maculate (DeGeer), Hippodamia convergens Guerin Meneville, Hippodamia parenthesis (Say), andHippodamia tredecimpunctata tibialis (Say)) and C. septempunctata comprised over 96 percent of all coccinellids collected. Of declining species, four Coccinella transversoguttata richardsoni Brown were collected from alfalfa, but Coccinella novemnotata Herbst and Adalia bipunctata (L.) were not found during the study. Coccinellid abundance was seldom affected by CMI. Coccinellids were more abundant in wheat-alfalfa in 1995 under high than low CMI. Coccinellid abundance in maize and alfalfa did not differ with CMI. A prey species, Empoasca fabae (Harris) (Heteroptera: Auchenorryncha: Cicadellidae), was more abundant in alfalfa in 1995 under high and intermediate than under low CMI. Coccinellid abundance was not correlated with that of E. fabae in 1995, and showed inconsistent association with E. fabae during the study. In grass, adult coccinellids (total across species), adult H. tredecimpunctata tibialis, and aphids were more abundant in warm- season grasses than in cool-season or mixed grass stands in one of three years. Our results provide further evidence that C. septempunctata has become relatively abundant in eastern South Dakota, whereas C. transversoguttata richardsoni, C. novemnotata, and A. bipunctata have become rare or absent

Electronic circuit implementation of chaos synchronization

In this paper, an electronic circuit implementation of a robustly chaotic two-dimensional map is presented. Two such electronic circuits are realized. One of the circuits is configured as the driver and the other circuit is configured as the driven system. Synchronization of chaos between the driver and the driven system is demonstrated

Causal sites as quantum geometry

We propose a structure called a causal site to use as a setting for quantum geometry, replacing the underlying point set. The structure has an interesting categorical form, and a natural "tangent 2-bundle," analogous to the tangent bundle of a smooth manifold. Examples with reasonable finiteness conditions have an intrinsic geometry, which can approximate classical solutions to general relativity. We propose an approach to quantization of causal sites as well.Comment: 21 pages, 3 eps figures; v2: added references; to appear in JM

A multiplex PCR assay for differentiating economically important gastrointestinal nematodes of cattle

A multiplex polymerase chain reaction (PCR) test was developed for identifying gastrointestinal (GI) nematodes that commonly infect cattle. This assay was developed using adult-derived genomic DNA and shown capable of discriminating parasite eggs from the feces of experimentally-infected animals at both the species and genus levels. Sequence data from internal (ITS) and external (ETS) transcribed spacers of the ribosomal DNA (rDNA) repeats as well as the 3′-end of the small subunit rDNA and 5′-end of the large subunit rDNA were used to generate five primer sets which, when used simultaneously in a multiplex PCR, produce a unique electrophoretic DNA banding pattern charac- terized by a single DNA fragment for Ostertagia ostertagi (257 bp), Haemonchus placei (176 bp),Oesophagostomum radiatum (329 bp), Trichostrongylus colubriformis (243 bp) and Cooperia on- cophora (151 bp). In a similar manner, the constructed primer sets amplified DNA from Ostertagia lyrata, Haemonchus contortus, Trichostrongylus axei, Cooperia surnabada and Cooperia punctata. With respect to H. contortus, a closely migrating doublet was generated suggesting size heterogene- ity in the ETS which is consistent with multiple rDNA repeat units within this species. PCR analyses using mixtures of monospecifically-purified nematode eggs indicated a sensitivity of less than 0.5 egg-DNA equivalent per species. Although, not designed as a quantitative technique, relative PCR signal intensities corresponded to relative egg burdens within the DNA samples from mixed species of eggs. Published by Elsevier Science B.V