Prognostic value of kallikrein-related peptidase 6 protein expression levels in advanced ovarian cancer evaluated by automated quantitative analysis (AQUA)

Kallikrein-related peptidases, a subgroup of the serine protease enzyme family, are considered important prognostic biomarkers in cancer. In the present study, we sought to determine the prognostic value of kallikrein-related peptidase 6 (KLK6) in ovarian cancer using a novel method of compartmentalized in situ protein analysis. A tissue array composed of 150 advanced stage ovarian cancers, uniformly treated with surgical debulking followed by platinum-paclitaxel combination chemotherapy, was constructed. For evaluation of KLK6 protein expression, we used an immunofluorescence-based method of automated in situ quantitative measurement of protein analysis (AQUA). Mean follow-up time of the cohort was 34.35 months. One hundred and thirty-five of 150 cases had sufficient tissue for AQUA analysis. In univariate survival analysis, low tumor KLK6 expression was associated with better outcome for overall survival over 3 years (P = 0.019). There was no association between tumor KLK6 expression and progression-free survival (P = 0.128). In multivariate survival analysis, adjusting for well-characterized prognostic variables, low tumor KLK6 expression level was one of the most significant predictor variable for overall survival (95% confidence interval, 1.19-3.50; P = 0.009). High tumor KLK6 protein expression is associated with inferior patient outcome in ovarian cancer. KLK6 may represent a promising disease biomarker and therapeutic target in ovarian cancer.     

Three new serum markers for prostate cancer detection within a percent free PSA-based artificial neural network

BACKGROUND: We aimed to evaluate the value of macrophage inhibitory cytokine 1 (MIC-1), human kallikrein 11 (hK11) migration inhibitor factor (MIF) in comparison to prostate-specific antigen (PSA) and % fPSA and also to develop a % fPSA-based ANN with the new input factors to determine whether these additional markers can further eliminate unnecessary prostate biopsies. METHODS: Serum samples from 371 patients with prostate cancer (PCa, n=135) or benign prostate hyperplasia (BPH, n=236) within the PSA range 0.5-20 microg/L were analyzed for total PSA, free PSA, MIC-1, hK11, and MIF. 'Leave one out' ANN models with these variables and prostate volume were constructed and compared to logistic regression (LR) and all single parameters. RESULTS: The discriminatory power of MIC-1, hK11, and MIF was less than that for PSA despite significant differences in BPH compared to PCa patients. At 90% and 95% sensitivity, the artificial neural networks (ANNs) were only significantly better than % fPSA if prostate volume was included. CONCLUSIONS: ANNs with the novel input factors of MIC-1, MIF, and/or hK11 and additional use of prostate volume demonstrated significant advantage compared with % fPSA and tPSA and may lead to a reduction in unnecessary prostate biopsies.     

The emerging roles of human tissue kallikreins in cancer

Human tissue kallikreins (hKs), which are encoded by the largest contiguous cluster of protease genes in the human genome, are secreted serine proteases with diverse expression patterns and physiological roles. Although primarily known for their clinical applicability as cancer biomarkers, recent evidence implicates hKs in many cancer-related processes, including cell-growth regulation, angiogenesis, invasion and metastasis. They have been shown to promote or inhibit neoplastic progression, acting individually and/or in cascades with other hKs and proteases, and might represent attractive targets for therapeutic intervention.     

Human tissue kallikreins: a family of new cancer biomarkers

Kallikreins are a subgroup of the serine protease enzyme family. Until recently, it was thought that the human kallikrein gene family contained only three members. In the past 3 years, the entire human kallikrein gene locus was discovered and found to contain 15 kallikrein genes. Kallikreins are expressed in many tissues, including steroid hormone-producing or hormone-dependent tissues such as the prostate, breast, ovary, and testis. Most, if not all, kallikreins are regulated by steroid hormones in cancer cell lines. There is strong but circumstantial evidence linking kallikreins and cancer. Prostate-specific antigen (PSA; hK3) and, more recently, human glandular kallikrein (hK2) are widely used tumor markers for prostate cancer. Three other kallikreins, hK6, hK10, and hK11, are emerging new serum biomarkers for ovarian and prostate cancer diagnosis and prognosis. Several other kallikreins are differentially expressed at both the mRNA and protein levels in various endocrine-related malignancies, and they have prognostic value. The coexpression of many kallikreins in the same tissues (healthy and malignant) points to the possible involvement of kallikreins in cascade enzymatic pathways. In addition to their diagnostic/prognostic potential, kallikreins may also emerge as attractive targets for therapeutics.     

Integrated stereotaxic imaging with CT, MR imaging, and digital subtraction angiography

A stereotaxic frame, compatible with digital subtraction angiography, computed tomography, and magnetic resonance imaging, is described along with a set of software programs that run in an independent imaging computer system, as well as in the computers associated with each modality. Plexiglas plates fastened to the sides of the frame contain fiducial markers that can be recognized in the images and from which the section position and in-plane coordinates of any point in the image relative to the frame may be determined. Coordinate measurements of isolated point targets may be made to an accuracy of better than +/- 1 mm within a 15-cm field of view in the plane of the section or projection on all modalities. The stereotaxic system is of sufficiently high accuracy to be used on a routine clinical basis with one or more of the above modalities.     

Human kallikrein 4: quantitative study in tissues and evidence for its secretion into biological fluids

BACKGROUND: Human kallikrein 4 (hK4) is a proteolytic enzyme belonging to the tissue kallikrein family of serine proteases. Previous tissue expression studies have demonstrated highest KLK4 mRNA expression in prostatic tissue, but there has been only limited evidence for the presence of hK4 protein in prostate and other tissues and in corresponding biological secretions. METHODS: To investigate the concentrations of hK4 in tissues and biological fluids, we developed a new hK4-specific sandwich-type immunoassay using a monoclonal antibody as the capture reagent. RESULTS: The assay has a detection limit of 0.02 microg/L and <0.1% cross-reactivity toward any of the other 14 human kallikreins. Twelve of 40 tissue extracts prepared from various human tissues contained detectable hK4 concentrations (0.68-7143 ng/g of total protein), with healthy prostate tissue containing the highest amount of hK4. Examination of 16 malignant and 18 benign prostate tissues revealed no significant differences in hK4 protein content, and the tissues contained a wide range of values (benign, <0.02 to 801 ng/g; malignant, <0.02 to 824 ng/g). Among the biological fluids tested, seminal plasma and urine contained widely varying amounts of hK4; concentrations in 54 urine samples were <0.02 to 2.6 microg/L, whereas concentrations in 58 seminal plasma samples were 0.2-202 microg/L. Affinity purification of hK4 from seminal plasma and subsequent mass spectrometry demonstrated the secreted nature of hK4 in seminal plasma. CONCLUSIONS: hK4 is found primarily in prostate tissue and is secreted in seminal plasma. Its value as a novel prostatic biomarker needs to be defined further.     

Human kallikrein 11: an indicator of favorable prognosis in ovarian cancer patients

OBJECTIVES: Human kallikrein 11 (hK11) is a secreted serine protease, highly expressed in hormonally regulated tissues, including the prostate and the ovary. Our preliminary studies indicate that hK11 may represent a diagnostic and prognostic biomarker for ovarian cancer. The aim of the present study was to examine the prognostic value of hK11 expression in ovarian tumors. METHODS: Using our established immunofluorometric assay, hK11 levels were quantified (ng per mg of total protein) in 134 ovarian tumor extracts and correlated with various clinicopathological variables and outcome [progression-free survival (PFS), overall survival (OS)], over a median follow-up period of 42 months. RESULTS: hK11 concentration in ovarian tumor cytosols ranged from 0 to 155 ng/mg of total protein, with a median of 1.45 ng/mg. An optimal cutoff value of 6.3 ng/mg was selected to categorize tumors as hK11-positive or negative. hK11-positive tumors were most often of early stage (Stage I/II) and grade (G1/G2) (P < 0.05). Univariate analysis revealed that patients with hK11-positive tumors had a significantly longer PFS (HR of 0.39, P = 0.005) and OS (HR of 0.44, P = 0.033). Cox multivariate analysis indicated that hK11 was an independent prognostic indicator of PFS (HR of 0.47, P = 0.042). Kaplan-Meier survival curves further confirmed that women with hK11-positive tumors have longer PFS and OS (P = 0.003 and P = 0.028, respectively). Also, a weak positive correlation was found between the expression levels of tissue hK11 and tissue CA125 (rs = 0.508; P < 0.001). CONCLUSIONS: These results further validate our initial findings that hK11 is an independent marker of favorable prognosis in ovarian cancer patients.     

A Relationship Between Fluctuations in Heart Rate and The Duration of Subsequent Episodes of Atrial Fibrillation

A relationship between autonomic tone and the onset of paroxysmal atrial fibrillation in some patients is recognised Episodes of PAF may vary enormously in duration, however, from a few beats to many hours. Whether autonomic tone influences the duration of ihe episodes has been less well investigated.
From a database of Holter recording taken from patients with symptomatic PAF, we identified all episodes of at least 30 seconds duration which were preceded by noise free sinus rhythm. This study examined the heart rate prior to AF onset, the change in heart rate over the final minute of sinus rhythm and the time of AF onset, and compared the data from those episodes of AF of more than 5 minutes duration to the shorter ones. Heart rate was slower before long episodes of AF, but this was found to predominantly represent data from separate recordings. A highly significant rise in heart rate was detected prior to long AF episodes compared to shorter ones. Daytime AF episodes were slightly longer than nocturnal ones.
The most important finding was that longer AF episodes were typified by a heart rate acceleration. This suggests that, regardless of underlying aetiology, and increase in sympathetic tone may be important in the sustenance of episodes of PAF.