Immunofluorometric quantitation and histochemical localisation of kallikrein 6 protein in ovarian cancer tissue: a new independent unfavourable prognostic biomarker.

Human kallikrein 6 protein is a newly discovered human kallikrein. We determined the amount of human kallikrein 6 in extracts of 182 ovarian tumours and correlated specific activity (ng hK6 mg(-1) total protein) with clinicopathological variables documented at the time of surgical excision and with outcome (progression free survival, overall survival) monitored over a median interval of 62 months. Thirty per cent of the tumours were positive for human kallikrein 6 (>35 ng hK6 mg(-1) total protein). Human kallikrein 6-specific immunohistochemical staining of four ovarian tissues that included benign, borderline and malignant lesions indicated a cytoplasmic location of human kallikrein 6 in tumour cells of epithelial origin, although the intensity of staining was variable. Tumour human kallikrein 6 (ng hK6 mg(-1) total protein) was higher in late stage disease, serous histotype, residual tumour >1 cm and suboptimal debulking (>1 cm) (P<0.05). Univariate analysis revealed that patients with tumour human kallikrein 6 positive specific activity were more likely to suffer progressive disease and to die (hazard ratio 1.71 (P=0.015) and 1.88 (P=0.022), respectively). Survival curves demonstrated the same (P=0.013 and 0.019, respectively). Multivariate analysis revealed that human kallikrein 6 positivity was retained as an independent prognostic variable in several subgroups of patients, namely those with (low) grade I and II tumours (hazard ratio progression free survival 4.3 (P=0.027) and overall survival 4.1 (P=0.023)) and those with optimal debulking (hazard ratio progression free survival 3.8 (P=0.019) and overall survival 5.6 (P=0.011)). We conclude that tumour kallikrein 6 protein levels have utility as an independent adverse prognostic marker in a subgroup of ovarian cancer patients with otherwise apparently good prognosis.

Breast cancer prognostic significance of a single nucleotide polymorphism in the proximal androgen response element of the prostate specific antigen gene promoter.

Prostate Specific Antigen (PSA) expression by breast epithelial cells is associated with favorable breast cancer prognosis. In preliminary studies, we found that a nucleotide variation (G-->A) at position -158 in the androgen response element (ARE-1) of the PSA promoter was present in four out of 9 breast tumors examined and in a breast carcinoma cell line. We have now determined the nucleotide composition at position -158 of DNA extracted from 148 well-characterized breast tumors and compared tumor genotype with that of controls without cancer, with tumor PSA concentration and with clinicopathological variables, overall survival and disease free survival. The G-->A base change at position -158 is a polymorphism. Allelotypes were similarly distributed in breast cancer patients and controls. The Mann-Whitney U Test showed a significantly higher tumor PSA concentration in tumors that presented a homozygous G as opposed to homozygous A genotype. Genotype at position -158 was not associated with clinicopathological variables in contingency table analysis. Univariate Cox regression models showed a 28% reduction in risk for death in patients with homozygous G genotype compared to those with homozygous A genotype (P = 0.03). However, ARE-1 genotype did not significantly add to the prognostic power in the multivariate model of overall survival. In summary, the base change at position -158 is a polymorphism that may affect breast cancer prognosis, but further studies are required to confirm this possibility and to investigate the relevance of this polymorphism in terms of breast cancer susceptibility.

Assay of prostate-specific antigen from whole blood spotted on filter paper and application to prostate cancer screening.

We report the measure of prostate-specific antigen (PSA) from extracts of blood dried on filter paper. Five 3-mm (diameter) paper discs containing approximately 25 microL of dried whole blood were punched from the filter paper and extracted with 500 microL of buffer. Recovery of PSA was > 92%. Imprecision of the filter paper procedure was <10% when corresponding whole-blood concentrations were >0.35 micrograms/L. PSA recovery was unaffected whether blood was applied to the filter as one 85-microL aliquots, two 43-microL aliquots, or three 28-microL aliquots. PSA is contained in the plasma fraction. Variation in hematocrit from 0.61 to 0.31 caused <+/-10% change in filter paper PSA. Regression analysis showed: filter paper PSA = 0.86 whole-blood PSA - 0.02; Sy/x = 0.44. Men (153) without prostate cancer gave a 95th percentile of 4.8 micrograms /L. PSA in filter paper dried blood was stable for >1 month at -20 to 37 degrees C and showed no loss of recovery after being mailed to a hot climate. We conclude that the filter paper procedure can reliably distinguish normal from increased concentrations of PSA and that it could facilitate screening to detect occult prostate cancer in large-scale mail-in programs to centralized laboratories.

Efficacy of feedback from quarterly laboratory comparison in maintaining quality of a hospital capillary blood glucose monitoring program.

OBJECTIVE: A 1-year randomized prospective study was conducted to investigate the efficacy of feedback from split-sample testing as part of a capillary blood glucose quality assurance program. RESEARCH DESIGN AND METHODS: A total of 124 nurses were randomized to either group A (quarterly comparisons with feedback) or group B (no feedback). The measure of nurse accuracy against the laboratory at 0, 6, and 12 months was determined by an additional five to seven split-sample tests without giving feedback to either group. Mean accuracy was determined in terms of percent absolute deviation from the laboratory result and a clinical consensus limit of +/- 20% deviation from the laboratory. RESULTS: By 12 months, there was a significant effect of feedback on nurse agreement with the laboratory method (P = 0.022 when agreement was scored as the mean percent absolute difference and P = 0.002 when agreement was scored in terms of the +/- 20% clinical consensus limit). Nurses in the group who had received no quarterly feedback from split-sample testing produced a 3.5% greater mean percent absolute deviation from the laboratory method and 12% fewer comparisons within the acceptable +/- 20% range. CONCLUSIONS: Feedback received from split-sample testing has a significant effect in maintaining accuracy in capillary blood glucose monitoring.

Effect of phorbol ester on the inhibition of proteoglycan synthesis induced by interleukin 1 and antiinflammatory drugs.

Low (2 ng/ml) and high (40 ng/ml) concentrations of the protein kinase C (PKC) activator phorbol-12-myristate-13-acetate (PMA) were tested for their effect on cultured bovine articular chondrocyte proteoglycan synthesis. In addition, we examined whether PMA could reverse interleukin 1 (IL-1) and nonsteroidal antiinflammatory drug (NSAID) induced inhibition of proteoglycan synthesis. Low concentrations of PMA stimulated proteoglycan synthesis by chondrocytes. High concentrations of PMA had no significant effect. IL-1 and high concentrations of NSAID inhibited proteoglycan production by chondrocytes. Low concentrations of PMA completely reversed IL-1 induced inhibition but did not significantly alter proteoglycan synthesis in the presence of antiinflammatory drugs. On the other hand, high concentrations of PMA had little effect on IL-1 induced inhibition but significantly potentiated the suppression of proteoglycan synthesis induced by 2 of the NSAID tested, indomethacin and flurbiprofen. Assay of PKC activity indicated that PKC levels were down-regulated by high but not by low concentrations of PMA. This suggests that different mechanisms were regulating the effects of low and high concentrations of PMA on proteoglycan synthesis. Although IL-1 and high concentrations of NSAID both suppress proteoglycan synthesis by chondrocytes, their different responses when coincubated with PMA suggest that they act through different pathways.