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

Anomalous Chiral Action from the Path-Integral

By generalizing the Fujikawa approach, we show in the path-integral formalism: (1) how the infinitesimal variation of the fermion measure can be integrated to obtain the full anomalous chiral action; (2) how the action derived in this way can be identified as the Chern-Simons term in five dimensions, if the anomaly is consistent; (3) how the regularization can be carried out, so as to lead to the consistent anomaly and not to the covariant anomaly. Our method uses Schwinger's ``proper-time'' representation of the Green's function and the gauge invariant point-splitting technique. We find that the consistency requirement and the point-splitting technique allow both an anomalous and a non-anomalous action. In the end, the nature of the vacuum determines whether we have an anomalous theory, or, a non-anomalous theoryComment: 28 page

Recovery of Viable Microorganisms from Solids. I - Model Systems

Solidification process for recovering viable microorganisms to aid spacecraft sterilization procedure

Inactivation of the major hemolysin gene influences expression of the nonribosomal peptide synthetase gene swrA in the insect pathogen Serratia sp. strain SCBI

Hemolysins are important virulence factors for many bacterial pathogens, including Serratia marcescens. The role of the major hemolysin gene in the insect pathogen Serratia sp. SCBI was investigated using both forward and reverse genetics approaches. Introduction of the major hemolysin gene into Escherichia coli resulted in a gain of both virulence and hemolytic activity. Inactivation of this hemolysin in Serratia sp. SCBI resulted in loss of hemolysis, but did not attenuate insecticidal activity. Unexpectedly, inactivation of the hemolysin gene in Serratia sp. SCBI resulted in significantly increased motility as well as increased antimicrobial activity. qRT-PCR analysis of mutants with a disrupted hemolysin gene showed a dramatic increase in mRNA levels of a nonribosomal peptide synthetase gene, swrA, which produces the surfactant serrawettin W2. Mutation of the swrA gene in Serratia sp. SCBI resulted in highly variable antibiotic activity, motility, virulence and hemolysis phenotypes that were dependent on the site of disruption within this 17.75 KB gene. When introduced into E. coli, swrA increases rates of motility and confers antimicrobial activity. While it is unclear how inactivation of the major hemolysin gene influences expression of swrA, these results suggest swrA plays an important role in motility and antimicrobial activity in Serratia sp. SCBI

Directed flow as effect of transient matter rotation in hadron and nucleus collisions

We discuss directed flow introduced for description of nucleus collisions and consider its possible behavior in hadronic and nuclei reactions due to rotation of the transient matter.Comment: 18 pages, 6 figure

How Sensitive are Di-Leptons from Rho Mesons to the High Baryon Density Region?

We show that the measurement of di-leptons might provide only a restricted view into the most dense stages of heavy ion reactions. Thus, possible studies of meson and baryon properties at high baryon densities, as e.g. done at GSI-HADES and envisioned for FAIR-CBM, might observe weaker effects than currently expected in certain approaches. We argue that the strong absorption of resonances in the high baryon density region of the heavy ion collision masks information from the early hot and dense phase due to a strong increase of the total decay width because of collisional broadening. To obtain additional information, we also compare the currently used approaches to extract di-leptons from transport simulations - i.e. shining, only vector mesons from final baryon resonance decays and instant emission of di-leptons and find a strong sensitivity on the method employed in particular at FAIR and SPS energies. It is shown explicitly that a restriction to rho meson (and therefore di-lepton) production only in final state baryon resonance decays provide a strong bias towards rather low baryon densities. The results presented are obtained from UrQMD v2.3 calculations using the standard set-up.Comment: 8 pages, 6 figures, expanded versio