BACKGROUND Brain death and cold preservation are major alloantigen-independent risk factors for transplantation outcome. The present study was conducted to assess the influence of these factors on transplantation-associated injury independently or in combination. METHODS Brain death was induced in F344 rats. Renal grafts were harvested after 6 hr and either directly transplanted in unilateral...

Saisho Y, Manesso E, Gurlo T, Huang CJ, Toffolo GM, Cobelli C, Butler PC. Development of factors to convert frequency to rate for -cell replication and apoptosis quantified by time-lapse video microscopy and immunohistochemistry. Am J Physiol Endocrinol Metab 296: E89 –E96, 2009. First published October 21, 2008; doi:10.1152/ajpendo.90697.2008.—An obstacle to development of methods to quantify ...

Polymerase chain reactions (PCRs) synthesize abundant amplification products. Contamination of new PCRs with trace amounts of these products, called carry-over contamination, yields false positive results. Carry-over contamination from some previous PCR can be a significant problem, due both to the abundance of PCR products, and to the ideal structure of the contaminant material for re-amplific...

Genome integrity requires well controlled cellular pools of nucleotides. dUTPases are responsible for regulating cellular dUTP levels and providing dUMP for dTTP biosynthesis. In Staphylococcus, phage dUTPases are also suggested to be involved in a moonlighting function regulating the expression of pathogenicity-island genes. Staphylococcal phage trimeric dUTPase sequences include a specific in...

In situ polymerase chain reaction (in situ PCR), which can detect a few copies of genes within a cell by amplifying the target gene, was developed to better understand the biological functions of tissues. In this study, we optimized the protocol conditions for the detection of X chromosome-linked phosphoglycerate kinase-1 (pgk-1) gene in paraffin-embedded sections of mouse reproductive organs. ...

Deoxyuridine triphosphate (dUTP) misincorporation and uracil misrepair have long been implicated in fluoropyrimidine-induced DNA damage; however, the enzymatic activities responsible for these lesions have not been previously identified as critical determinants of overall sensitivity to the antitumor effects of these agents. The purpose of this study was to determine whether differences in urac...

INTRODUCTION This study was designed to quantify the role of the pericellular matrix (PCM) in chondrocyte apoptosis using chondrons, which are a cartilage functional unit including a chondrocyte and its associated PCM. METHODS Chondrocytes and chondrons were enzymatically isolated from human articular cartilage and exposed to monosodium iodoacetate (MIA) and staurosporine for apoptosis induct...

Objectives . This study investigated the substrate properties of modified derivatives triphosphates purine and pyrimidine deoxynucleosides (5-propynyl-2’-deoxyuridine-5’-triphosphate, 5-propynyl2’-deoxycytidine-5’-triphosphate, 5-methyl-2’-deoxycytidine-5’-triphosphate, N6-methyl-2’-deoxyadenosine-5’-triphosphate) during their simultaneous incorporation in enzymatic reactions (polymerase chain ...

OBJECTIVE To investigate DNA fragmentation by using terminal deoxyribonucleotidyl transferase-mediated dUTP nick-end labeling in relation to World Health Organization parameters and computer-aided sperm analysis (CASA) in sperm to determine the possibility of obtaining a correlation among CASA parameters, sperm morphology, and DNA fragmentation. DESIGN Sperm analysis according to World Health...

Using a highly tumorigenic human breast cancer model (Ha-ras-transfected MCF7 cell line) we analyzed the efficacy of the differentiation-inducing agent sodium phenylacetate (NaPA), both in vitro and in vivo. NaPA-treated MCF7ras cells showed dose-dependent growth inhibition from 2.5 to 15 mM without apparent toxicity. Western blot analysis showed a Bcl-2 down-regulation after 48 h treatment wit...