Abstract. Polyclonal isoenzyme-specific antisera were developed against four calcium-independent protein kinase C (PKC) isoenzymes (S, e, e', and~) as well as the calcium-dependent isoforms (a, 01, ßn, and -y).These antisera showed high specificities, high titers, and high binding affinities (3-370 nM) for the peptide antigens to which they were raised . Each antiserum detected a species of the predicted molecular weight by Western blot that could be blocked with the immunizing peptide . PKC was sequentially purified from rat brain, and the calcium-dependent forms were finally resolved by hydroxyapatite chromatography . Peak I reacted exclusively with antisera to PKCy, peak II with PKCß, and -ßu, and peak III with PKCa. These same fractions, however, were devoid of immunoreactivity for the calcium-independent isoenzymes. The PKC isoenzymes demonstrated a distinc-P ROTEIN kinase C (PKC)' plays a major role in transmembrane signal transduction (35). PKC is activated by diacylglycerol, which is generated from membrane phospholipids upon stimulation of cells with various agonists (4) . PKC also serves as the receptor for phorbol esters and related tumor promoters (34), which activate the kinase by substituting for endogenous diacylglycerol (8) . Because of the pleiotropic actions of diacylglycerol and phorbol esters, PKC has been implicated in the regulation ofa variety of cellular processes, including proliferation, differentiation, and release of hormones and neurotransmitters (35,36).Molecular cloning studies have revealed that PKC consists of a large family of at least eight different isoenzymes (36, 37) that can be divided into two major groups. Initially, four isoforms of PKC were described . This group consists of PKCca, -01, -ß, 1, and -y, with PKCß, and -ß arising via alternate splicing of the same gene transcript and resulting in distinct carboxy-terminal regions . All four of these isoen-1. Abbreviations used in this paper: PKC, protein kinase C.O The Rockefeller University Press,

Objective To determine the accuracy of tumour associated antigens CA 125, CA 15–3 and TAG 72.3 in the differential diagnosis of benign and malignant pelvic masses and to compare the results with a previously defined risk of malignancy index (RMI). Design Retrospective analysis of samples collected during a prospective observational study. Setting Department of Obstetrics and Gynaecology, the Royal London Hospital and Duke University Medical Center. Subjects One hundred and forty‐three consecutive patients undergoing surgery for an adnexal mass. Method Tumour marker levels were determined by radio‐immunoassay in stored serum samples obtained from 143 study subjects. Results The highest diagnostic accuracy of the tumour marker panel was achieved by defining a positive result as elevation of any two of CA 125 (>30 u/ml), CA 15–3 (>30 u/ml) and TAG 72.3 (> 10 u/ml), (sensitivity 66.7%, specificity 93.1%). Similar diagnostic accuracy could be achieved by CA 125 alone using an upper limit of 50 u/ml (sensitivity 66.7%, specificity 94.1%). Inclusion of CA 15–3 or TAG 72.3 in stepwise logistic regression analysis did not improve the discriminative performance of the RMI. Conclusion The risk of malignancy index incorporating CA 125, menopausal status and ultrasound is superior to the panel of three tumour markers for pre‐operative differential diagnosis of the pelvic mass.

Serum levels and FE of calcium, phosphorus, magnesium and uric acid were significantly higher in CKD patients compared to healthy very old people with similar GFR, except for serum magnesium and FE of uric acid, which were similar in both groups.

No abstract

The solid tumor mRNA expression of genes related to the mechanism of action of certain antineoplastic agents is often predictive of clinical efficacy. We report here on the development of a rapid and practical real-time RT-PCR method to quantify genetic expression in solid tumors. The genes examined are related to the intracellular pharmacology of gemcitabine and cisplatin, two drugs that are used in the treatment of several types of advanced cancer. We evaluated target gene mRNA levels from breast tumor samples using two quantitative RT-PCR methods: 1) an improved relative RT-PCR method using fluorescence-labeled primers, automated PCR set up, and GeneScan analysis software; and 2) real-time RT-PCR with redesigned primers using an ABI 7900HT instrument, with additional postprocessing of the data to adjust for efficiency differences across the target genes. Using these methods, we quantified mRNA expression levels of deoxycytidine kinase (dCK), deoxycytidylate deaminase (dCDA), the M1 and M2 subunits of ribonucleotide reductase (RRM1, RRM2), and excision cross complementation group 1 (ERCC1) in 35 human "fresh" frozen breast cancer biopsies. While both assay methods were substantially more rapid than traditional RT-PCR, real-time RT-PCR appeared to be superior to the amplification end-point measurement in terms of precision and high throughput, even when a DNA sequencer was used to assess fluorescence-labeled PCR products. This reproducible, highly sensitive real-time RT-PCR method for the detection and quantification of the mRNAs for dCK, dCDA, RRM1, RRM2, and ERCC1 in human breast cancer biopsies appears to be more informative and less time-consuming than either classical radioisotope-dependent RT-PCR or the technique utilizing GeneScan analysis described herein. By allowing the measurement of intratumoral target gene expression, these new methods may prove useful in predicting the clinical utility of gemcitabine- and platinum-containing chemotherapy programs in patients with solid tumors.