Erwinia amylovora secretes DspE, a pathogenicity factor and functional AvrE homolog, through the Hrp (type III secretion) pathway

Erwinia amylovora was shown to secrete DspE, a pathogenicity factor of 198 kDa and a functional homolog of AvrE of Pseudomonas syringae pv. tomato. DspE was identified among the supernatant proteins isolated from cultures grown in an hrp gene-inducing minimal medium by immunodetection with a DspE-specific antiserum. Secretion required an intact Hrp pathway.     

Meprin A, the major matrix degrading enzyme in renal tubules, produces a novel nidogen fragment in vitro and in vivo

We examined the effect of meprin A, the major matrix degrading metalloproteinase in rat kidney, on the laminin-nidogen complex. N-terminal sequence information from the most abundant 55 kDa fragment revealed that it was a breakdown product of nidogen rather than laminin. In comparison with over 50 nidogen cleavage sites produced by other proteases, the meprin A-induced nidogen cleavage site at amino acid position 899-900, a glutamine-glycine site in the G3 domain, is unique. In addition, these data demonstrate that meprin A degrades the G3 domain of nidogen even in the presence of laminin binding, which usually accords protection from proteolytic degradation. Meprin A also degraded purified nidogen into similar breakdown products. Given that the tubular basement membrane is located on the basilar side of the cell, the location of meprin A on the apical brush border makes it difficult to envision a role for meprin A in injury-induced basement membrane component breakdown. Thus, we examined the possibility that following renal tubular epithelial cell injury, meprin A undergoes a translocation to reach the underlying basement membrane. After renal ischemia-reperfusion there was a marked alteration in meprin A staining with meprin A now distributed throughout the renal tubular cell cytoplasm and directly adherent to the tubular basement membrane. This was in contrast to the usual linear staining of the brush border of tubules in the corticomedullary junction. These data provide unequivocal evidence that following injury, meprin A undergoes redistribution and/or adherence to the tubular basement membrane. Since in our in vitro studies, we identified a distinct meprin-induced 55 kDa nidogen breakdown product, the urine was also examined for the presence of nidogen degradation products after rat renal ischemia-reperfusion injury. Western blots showed a marked increase in the urinary 55 kDa nidogen fragment as early as the first day following ischemia-reperfusion injury and continuing for six days. Taken together, these in vivo data strongly support the notion that the nidogen breakdown products are the result of partial degradation of tubular basement membrane by meprin A following renal tubular ischemia-reperfusion injury.     

Gender-related differences in susceptibility of A/J mouse to benzo[a]pyrene-induced pulmonary and forestomach tumorigenesis

A commercial patient dose verification system utilizing non-invasive metal oxide semiconductor field effect transistor (MOSFET) dosimeters originally designed for radiotherapy applications has been evaluated for use at diagnostic energy levels. The system features multiple dosimeters that may be used to monitor entrance or exit skin dose and intracavity doses in phantoms in real time. We have characterized both the standard MOSFET dosimeter designed for radiotherapy dose verification and a newly developed "high sensitivity" MOSFET dosimeter designed for lower dose measurements. The sensitivity, linearity, angular response, post-exposure response, and physical characteristics were evaluated. The average sensitivity (free in air, including backscatter) of the radiotherapy MOSFET dosimeters ranged from 3.55 x 10(4) mV per C kg(-1) (9.2 mV R(-1)) to 4.87 x 10(4) mV per C kg(-1) (12.6 mV R(-1)) depending on the energy of the x-ray field. The sensitivity of the "high sensitivity" MOSFET dosimeters ranged from 1.15 x 10(5) mV per C kg(-1) (29.7 mV R(-1)) to 1.38 x 10(5) mV per C kg(-1) (35.7 mV R(-1)) depending on the energy of the x-ray field. The high sensitivity dosimeters demonstrated excellent linearity at high energies (90 and 120 kVp) and acceptable linearity at lower energies (60 kVp). The angular response was significant for free-in-air exposures, as illustrated by the sensitivity differences between the two sides of the dosimeter, but was excellent for measurements within a tissue equivalent cylinder. The post-exposure drift response is a complicated but reproducible function of time. Real-time monitoring requires little if any corrections for the post-exposure drift response. The MOSFET dosimeter system brings some unique capabilities to diagnostic radiology dosimetry including small size, real-time capabilities, nondestructive measurement, good linearity, and a predictable angular response.     

Structural studies of ribosomal proteins

Crystal and solution structures of fourteen ribosomal proteins from thermophilic bacteria have been determined during the last decade. This paper reviews structural studies of ribosomal proteins from Thermus thermophilus carried out at the Institute of Protein Research (Pushchino, Russia) in collaboration with the University of Lund (Lund, Sweden) and the Center of Structural Biochemistry (Karolinska Institute, Huddinge, Sweden). New experimental data on the crystal structure of the ribosomal protein L30 from T. thermophilus are also included.     

Changes in the orientation of the smooth-muscle cells of the tunica media from major arteries under constant lengthwise stretching in situ

It has been shown that changes in the orientation of arterial smooth muscle cells during a constant longitudinal stretching of the artery in vivo are not similar in different sections of the stretching zone. Cells in the proximal and distal sections keep their orientation but this orientation differs from that of smooth muscle cells in the control arteries. Cells in the central part of the stretching region lose their definite orientation to settle randomly.     

Structure-activity relationship homology (SARAH): a conceptual framework for drug discovery in the genomic era

Extension of the traditional pharmacological approach of protein target classification to whole target systems has the potential to relate elements of protein sequence to the structure-activity relationship (SAR) of small molecules that can modulate protein action. Grouping potential drug discovery targets into families based on the relatedness of their SAR provides a means to translate the information from genome-sequencing efforts into knowledge that will aid in the discovery of drugs.     

Dental enamel hypoplasia in children with combined congenital and hereditary defects in the development of the CNS and the locomotor system (infantile cerebral palsy, spinal cord hernias and myopathies)]

The incidence of hypoplasia in children with congenital and hereditary developmental defects of the central nervous system and the locomotor system is high (44.5 +/- 3.5%), much higher than the incidence of hypoplasia in children without neurological disorders (2.0 +/- 2.0%). This is explained by exposure of the fetus and newborn to numerous intensive factors complicating the biological anamnesis because of profound disturbances of the metabolic processes and largely responsible for the underlying neurological disease and defects of hard dental tissues presenting as hypoplasia of the enamel.     

Role of caspases (ICE/CED 3 proteases) in DNA damage and cell death in response to a mitochondrial inhibitor, antimycin A

Caspases (ICE/ Ced3 proteases) are a closely related family of cysteine proteases that play a key role in apoptotic cell death. We examined the role of caspases in DNA damage and cell death in response to the mitochondrial inhibitor, antimycin A. LLC-PK1 cells contain caspase activity that was markedly inhibited by cleavage site-based peptide inhibitors of caspases but not by inhibitors of serine, cysteine, aspartate or metalloproteinases. The caspase activity increased within five minutes of exposure to antimycin A, preceding any evidence of DNA damage and cell death. The specific caspase inhibitors. Ac-Tyr-Val-Ala-Asp-aldehyde (inhibitor I) and Ac-Asp-Glu-Val-Asp-aldehyde (inhibitor II) prevented, in a dose dependent manner, antimycin A-induced DNA strand breaks as determined by DNA unwinding assay (residual double stranded DNA in control, 94 +/- 2%; antimycin A alone, 48 +/- 3%; antimycin A + inhibitor I at 50 microM, 93 +/- 2%; antimycin A + inhibitor II at 50 microM, 89 +/- 5%; N = 3 to 4, P < 0.001). These inhibitors also prevented antimycin A-induced DNA fragmentation as determined by agarose gel electrophoresis and by in situ labeling of cell nuclei by the terminal deoxynucleotidyl transferase (TdT) nick end labeling (TUNEL) method. The caspase inhibitors markedly prevented antimycin A-induced cell death in a dose-dependent manner as measured by trypan blue exclusion (control 6 +/- 1%, antimycin A alone 40 +/- 1%, antimycin A + inhibitor I at 50 microM 16 +/- 1%, antimycin A + inhibitor II at 50 microM 16 +/- 1%; N = 4 to 7, P < 0.001). These data indicate that the caspase family of enzymes play an important role in DNA damage and cell death in response to the mitochondrial inhibitor, antimycin A.