Removal of the catalytic subunit of DNA-protein kinase in the proximal tubules promotes DNA and tubular damage during kidney injury.

Tubular epithelial cell damage can be repaired through a series of complex signaling pathways. An early event in many forms of tubular damage is the observation of DNA damage, which can be repaired by specific pathways depending upon the type of genomic alteration.. In this study, we report that the catalytic subunit of DNA protein kinase (DNA-PKcs), a central DNA repair enzyme involved in sensing DNA damage and performing double stranded DNA break repair, plays an important role in the extent of tubular epithelial cell damage following exposure to injurious acute and chronic stimuli. Selective loss of DNA-PKcs in the proximal tubules led to increased markers of kidney dysfunction, DNA damage, and tubular epithelial cell injury in multiple models of acute kidney injury, specifically bilateral renal ischemia-reperfusion injury and single dose of cisplatin (15 mg/kg IP). In contrast, in a mouse model of kidney fibrosis and chronic kidney disease (UUO),the protective effects of DNA-PKcs was not as obvious histologically from the tissue sections. In the absence of proximal tubular DNA-PKcs, there was reduced levels of fibrotic markers, α-SMA and fibronectin, which suggests that there may be a biphasic role of DNA-PKcs depending upon the conditions exerted upon the kidney. In conclusion, this study demonstrates that the catalytic subunit of DNA-PKcs plays a context-dependent role in the kidney to reduce DNA damage during exposure to various types of acute, but not chronic forms of injurious stimuli.

The mechanisms of complement activation in normal bovine serum and normal horse serum against Yersinia enterocolitica O:9 strains with different outer membrane proteins content.

Yersinia enterocolitica is a common zoonotic pathogen and facultative intracellular bacterium which can survive within blood cells. Cattle and horses are considered a reservoir of Y. enterocolitica which often causes several serious syndromes associated with yersiniosis such as abortions, premature births or infertility. The aim of our investigation was to determine the vitality of Y. enterocolitica O:9 strains (Ye9) in bovine and horse sera (NBS and NHrS) and explain the role of outer membrane proteins (OMPs) in serum resistance of these bacteria. Our previous studies demonstrated moderate human serum (NHS) resistance of the wild type Ye9 strain, whereas mutants lacking YadA, Ail or OmpC remained sensitive to the bactericidal activity of NHS. The present study showed that the wild type of Ye9 strain was resistant to the bactericidal activity of both NHrS and NBS, while Ye9 mutants lacking the YadA, Ail and OmpC proteins were sensitive to NHrS and NBS as well as to NHS. The mechanisms of complement activation against Ye9 strains lacking Ail and YadA were distinguished, i.e. activation of the classical/lectin pathways decisive in the bactericidal mechanism of complement activation of NBS, parallel activation of the classical/lectin and alternative pathways of NHrS. In this research the mechanism of independent activation of the classical/lectin or the alternative pathway of NBS and NHrS against Ye9 lacking OmpC porin was also established. The results indicate that serum resistance of Ye9 is multifactorial, in which extracellular structures, i.e. outer membrane proteins (OMPs) such as Ail, OmpC or YadA, play the main role.

Instrumentation of broadband frequency domain thermoreflectance for measuring thermal conductivity accumulation functions.

This paper describes the instrumentation for broadband frequency domain thermoreflectance (BB-FDTR), a novel, continuous wave laser technique for measuring the thermal conductivity accumulation function. The thermal conductivity accumulation function describes cumulative contributions to the bulk thermal conductivity of a material from energy carriers with different mean free paths. It can be used to map reductions in thermal conductivity in nano-devices, which arise when the dimensions of the device are commensurate to the mean free path of energy carriers. BB-FDTR uses high frequency surface temperature modulation to generate non-diffusive phonon transport realized through a reduction in the perceived thermal conductivity. By controlling the modulation frequency it is possible to reconstruct the thermal conductivity accumulation function. A unique heterodyning technique is used to down-convert the signal, therein improving our signal to noise ratio and enabling results over a broader range of modulation frequencies (200 kHz-200 MHz) and hence mean free paths.

Stenotic glomerulotubular necks in radiation nephropathy.

Fibrosis of the renal interstitium is well correlated with kidney function and a greater extent of fibrosis predicts renal failure. Recent work has shown striking fibrotic constrictions at the glomerulotubular neck in porcine radiation nephropathy and in Wegener's granulomatosis in man. The present studies were designed to identify stenotic necks in a third species and to evaluate the effect of captopril treatment. Experimental radiation nephropathy was established with 17 Gy total body irradiation of barrier-maintained rats. Kidneys were obtained at 99 and 203 days for histology, using perfusion fixation. There was renal injury, with a rise in BUN, as expected, which was attenuated by captopril treatment. There was stenotic neck formation at 99 and 203 days. Captopril did not influence the absolute fraction of necks that were stenotic but it did prevent the evolution of glomeruli with necks to atubular glomeruli. It is concluded that stenosis of the glomerulotubular neck is a general phenomenon of any scarring kidney disease; that stenotic necks are probably an intermediate step in the evolution towards atubular glomeruli; and that timely use of captopril may prevent the progression of a stenotic neck towards an atubular glomerulus.