Predicting prostate biopsy outcome: prostate health index (phi) and prostate cancer antigen 3 (PCA3) are useful biomarkers.

Indication for prostate biopsy is presently mainly based on prostate-specific antigen (PSA) serum levels and digital-rectal examination (DRE). In view of the unsatisfactory accuracy of these two diagnostic exams, research has focused on novel markers to improve pre-biopsy prostate cancer detection, such as phi and PCA3. The purpose of this prospective study was to assess the diagnostic accuracy of phi and PCA3 for prostate cancer using biopsy as gold standard. Phi index (Beckman coulter immunoassay), PCA3 score (Progensa PCA3 assay) and other established biomarkers (tPSA, fPSA and %fPSA) were assessed before a 18-core prostate biopsy in a group of 251 subjects at their first biopsy. Values of %p2PSA and phi were significantly higher in patients with PCa compared with PCa-negative group (p<0.001) and also compared with high grade prostatic intraepithelial neoplasia (HGPIN) (p<0.001). PCA3 score values were significantly higher in PCa compared with PCa-negative subjects (p<0.001) and in HGPIN vs PCa-negative patients (p<0.001). ROC curve analysis showed that %p2PSA, phi and PCA3 are predictive of malignancy. In conclusion, %p2PSA, phi and PCA3 may predict a diagnosis of PCa in men undergoing their first prostate biopsy. PCA3 score is more useful in discriminating between HGPIN and non-cancer.

Preoperative insulin-like growth factor-binding protein-3 (IGFBP-3) blood level predicts gleason sum upgrading.

BACKGROUND: About 43% of men with low Gleason grade prostate cancer (PCa) at biopsy will be finally diagnosed with high-grade PCa at radical prostatectomy (RP). Gleason sum at RP is a good indicator of biochemical recurrence and poor clinical outcome. Therefore, there is a need to improve clinical evaluation of PCa aggressiveness in order to choice appropriate treatment. To this aim an easy-available tool is represented by circulating biomarkers. Among these, the best candidates are some molecules involved in PCa pathogenesis such as IGFBP-2 and IGFBP-3, IL-6, and its soluble receptor (SIL-6R). METHODS: In this study, we evaluated the ability of preoperative IGFBP-2, IGFBP-3, IL-6, and SIL-6R serum levels to predict Gleason score upgrade in 52 PCa patients. RESULTS: We found that IGFBP-3 median levels were significantly lower in patients who showed Gleason upgrading from biopsy to RP (P = 0.024). We also found an association between biopsy T-stage and Gleason Upgrade (P = 0.011). Using multivariate logistic regression model, we demonstrated that the association of IGFBP-3 serum levels together with biopsy T-stage and biopsy Gleason score was useful to calculate a prognostic risk score. ROC curve analysis of risk score showed a good ability to predict GSU (AUC = 0.81; 95% CI 0.69-0.93). CONCLUSIONS: Our results suggest that preoperative IGFBP-3 circulating levels determination may be useful to predict Gleason score upgrading alone and/or in combination with biopsy T-stage and biopsy Gleason score.

Soluble interleukin-6 receptor to interleukin-6 (sIL­6R/IL-6) ratio in serum as a predictor of high Gleason sum at radical prostatectomy.

Approximately 40% of males with low Gleason grade clinically localized prostate cancer (PCa) at biopsy were finally diagnosed with high Gleason grade PCa at radical prostatectomy (RP). Therefore, a more reliable assessment of the Gleason grade prior to RP is required. Readily available modalities such as circulating biomarkers may be useful for this purpose. The aim of this study was to evaluate the ability of preoperative interleukin 6 (IL­6) and its soluble receptor (sIL­6R), as well as urokinase-type plasminogen activator (u-PA), its receptor (u-PAR) and the inhibitor (PAI-1) to predict Gleason score upgrading. A total of 51 PCa patients with biopsy Gleason score ≤7 were studied. IL­6 and sIL­6R, uPA, uPAR and PAI-1 preoperative serum levels were determined. Differences in the median and mean values of the preoperative blood levels of all biomarkers between patients with and without Gleason score upgrading were tested. The prognostic performance of each biomarker was further assessed by means of receiver operating characteristic (ROC) curves. The results showed the sIL­6R and sIL­6R/IL-6 ratio median levels to be significantly higher in patients who had Gleason score upgrading from ≤7 at biopsy to >7 at RP (p=0.024 and p=0.011, respectively). The ROC curve revealed that sIL­6R and the sIL­6R/IL­6 ratio identified subjects at a high risk of upgrading [area under curve (AUC)=0.80 and AUC=0.83, respectively] with similar sensitivity and higher specificity for the ratio. The findings suggest that preoperative sIL­6R and sIL­6R/IL­6 ratio determination in serum are useful as prognostic biomarkers in PCa patients.

Matrix metalloproteinase-2 and -9 in the urine of prostate cancer patients.

The matrix metalloproteinase family of enzymes is comprised of critically important extracellular proteases whose activity has been implicated in a number of key normal and pathological processes. The latter include growth, progression and metastasis as well as dysregulated angiogenesis that is associated with these events. The MMPs are secreted by all types of cells, and they also carve through the extracellular matrix, allowing cancer cells to take root and metastasize. Endogenous inhibitors typically hold MMPs in check but in cancer, the balance shifts against the inhibitors and in favor of MMPs, which ultimately spill over from blood into urine. By gelatin zymography we verified MMP activity in concentrated urine of patients with prostate disease. Of these patients, 30 had cancer, consisting of 13 with Gleason score 6, 12 with Gleason 7, 2 with Gleason 8, 3 with Gleason 9 and 8 had benign prostate hyperplasia. Zymography showed 4 dominant gelatinolityc bands of 240, 130, 92 and 72 kDa in prostate disease. The most abundant lytic activity is at 92 kDa (MMP-9), whereas MMP-2 is present in lesser quantities. Moreover, MMP-9 activity is enhanced in the urine from patients with benign prostate hyperplasia compared with cancer patients. No correlation between gelatinolytic activity and Gleason score or pathological findings was found.

Effects of the ErbB1/ErbB2 kinase inhibitor GW2974 on androgen-independent prostate cancer PC-3 cell line growth and NSE, chromogranin A and osteopontin content.

Prostate cancer is one of the most frequently diagnosed cancer in men. Treatment by radical prostatectomy, radiotherapy and anti-androgen drugs is successful in patients with localized cancer. However, prolonged androgen deprivation often leads to hormone refractory condition, associated with disease relapse. ErbB1 and ErbB2 activity has been correlated with androgen-independence. We determined the effects of GW2974, a dual inhibitor of ErbB-1 and ErbB-2 tyrosine kinase activity, on growth, NSE, chromogranin A and osteopontin cytosol content in the androgen-independent prostate cancer cell line PC-3. We found that PC-3 cell growth was inhibited by GW2974, whereas NSE and chromogranin A cell contents were stimulated and osteopontin cytosol level was not affected. The present data may have clinical implications for the treatment of advanced prostate cancer.

Diagnostic value of carbohydrate antigens in supernatants and sediments of pleural effusions.

A panel of tumour markers including carcinoembryonic antigen (CEA), carbohydrate antigen (Ca)15-3, Ca125 and Ca19-9 were measured in the lysate of sediments and in the supernatants of pleural effusions of patients with benign and malignant disease. The tumour markers were also measured in the serum of the same patients. Of these patients, 32 had benign diseases (12 trasudative effusions associated with cirrhosis and 20 with non-malignant exudates: 12 pleuritis and 8 other inflammations) and 103 had malignant effusions (37 breast cancers, 29 lung cancers, 10 ovary cancers, 6 kidney cancers, 11 mesotheliomas and 10 lymphomas). We showed the highest level of CEA in pleural effusions of lung cancer followed by that in pleural effusions of breast cancer; whereas Ca15-3 was very high in the pleural effusions of breast and lung cancer. Concerning the lysate of sediment, CEA was high in the pleural effusions of patients with lung cancer and Ca15-3 in those of patients with breast cancer. The other markers are much less useful. For the remaining tumours, none of the markers tested appear to aid in the diagnosis of disease. In conclusion, our data suggest that the combined determination of tumour markers on supernatants and sediments of pleural effusion may provide additional information on the nature of pleural effusion, especially for cases with negative cytology.

Iodine status assessment in Campania (Italy) as determined by urinary iodine excretion.

OBJECTIVE: Mild iodine deficiency was first documented in Campania in the 1990s. We assessed the urinary iodine nutritional status of schoolchildren in Campania before the introduction of legislation for salt iodization and compared the findings with previous results to evaluate to what extent "silent" iodine prophylaxis, which accompanies socioeconomic advances, affects iodine status. METHODS: We examined 10552 schoolchildren aged 9-13 y from the five Campania provinces. The study was conducted from April 1999 to October 2002. Urinary iodine excretion was measured in morning urine samples with the AutoAnalyzer 3, an automated system based on the Sandell-Kolthoff reaction. Data were interpreted according to World Health Organization criteria. RESULTS: The median urinary iodine excretion level in Campania was less than 100 micromicrog/L, which indicates insufficient iodine intake. Mild iodine deficiency was identified in all provinces, namely Napoli, Salerno, Caserta, Avellino, and Benevento, with median urinary iodine excretions of 87, 81, 72, 64, and 61 microg/L, respectively. Overall, the analysis of frequency distribution showed values below 50 and 100 microg/L in 32% and 61% of children, respectively. These values were lower than those previously reported for Campania. CONCLUSION: This study confirms that Campania is a mild iodine deficiency area. The decrease in iodine deficiency versus previous studies indicates that silent prophylaxis plays a relevant role in this condition, but it is not sufficient to eradicate it. Our data will serve as a basis for future evaluations of iodine status in Campania.

The contribution of omental adipose tissue to adipokine concentrations in patients with the metabolic syndrome.

PURPOSE: To examine differences in peripheral vascular endothelial growth factor (VEGF), interleukin-6 (IL6) and cortisol concentrations between patients with both visceral obesity and metabolic syndrome, and lean controls. In a subsample of metabolic patients underwent abdominal surgery, the adipokine concentrations were measured in venous blood from the omentum to determine information on some processes of synthesis. METHODS: Forty-two healthy lean controls and 46 overweight-obese patients with central adiposity and stigmata of metabolic syndrome were studied. In a subsample of 11 metabolic patients undergoing non-bariatric surgery, blood samples from omental and peripheral veins were taken intraoperatively to determine VEGF, IL6 and cortisol concentrations. RESULTS: Median levels (range) of peripheral VEGF and IL6 were higher in patients than in controls [31.5 (3-112) pg/mL vs 21.35 (9-41.9) pg/mL (P < 0.05) and 5.50 (1.40-13) pg/mL vs 1.15 (0.3-1) pg/mL (P < 0.0001)]. On the other hand, concentrations of VEGF and IL6 from the omental and peripheral veins were similar in the surgery sub-group. Peripheral cortisol concentrations were not higher in patients than in controls, nor were omental concentrations different from the peripheral. Omental and peripheral VEGF and cortisol values were correlated, whereas no association was found between omental and peripheral IL6. CONCLUSIONS: In the presence of abdominal obesity, VEGF and IL6 concentrations are increased in the systemic circulation. The contribution of visceral adipose tissue to circulating levels of VEGF and IL6 was modest.

Gelatinolytic activities (matrix metalloproteinase-2 and -9) and soluble extracellular domain of Her-2/neu in pleural effusions.

Matrix metalloproteinases (MMPs) are proteolytic enzymes that are implicated in multiple stages of cancer progression including invasion and metastasis. MMPs exert these effects by cleaving a diverse group of substrates, which include not only structural components of the extracellular matrix, but also growth factor receptors. By gelatin zymography we verified MMP activity in the pleural effusions of patients with benign and malignant disease. Of these patients, 32 had malignant pleural effusion, consisting of 20 breast cancer, 6 non-small cell lung carcinoma, 4 ovarian carcinoma, and 2 colonic adenocarcinoma, and 10 had benign pleural effusion (5 pleurisy and 5 cirrhosis). Zymography showed the constant presence of a substantial amount of MMP-2 in all samples analyzed, whereas MMP-9 was present to lesser quantities. MMP-2 activity was enhanced in pleural effusions from patients with benign diseases compared with cancer patients. MMP-9 was present in 59% of cancer patients and the lytic activity was enhanced in pleurisy and absent in cirrhosis. Furthermore, we determined the pleural effusion levels of the soluble extracellular domain of HER-2/neu. The levels of HER-2/neu ECD were above the cut-off value in breast cancer patients. No correlation between gelatinolytic activities and high HER-2/neu ECD values was found.

Acute effects of triiodothyronine on endothelial function in human subjects.

CONTEXT: Thyroid hormone regulates several cardiovascular functions, and low T(3) levels are frequently associated with cardiovascular diseases. Whether T(3) exerts any acute and direct effect on endothelial function in humans is unknown. OBJECTIVE: Our objective was to clarify whether acute changes in serum T3 concentration affect endothelial function. DESIGN, SETTING, AND SUBJECTS: Ten healthy subjects (age, 24 +/- 1 yr) participated in a double-blind, placebo-controlled trial at a university hospital. INTERVENTIONS: T3 (or placebo) was infused for 7 h into the brachial artery to raise local T3 to levels observed in moderate hyperthyroidism. Vascular reactivity was tested by intraarterial infusion of vasoactive agents. MAIN OUTCOME MEASURES: We assessed changes in forearm blood flow (FBF) measured by plethysmography. RESULTS: FBF response to the endothelium-dependent vasodilator acetylcholine was enhanced by T3 (P = 0.002 for the interaction between T3 and acetylcholine). The slopes of the dose-response curves were 0.41 +/- 0.06 and 0.23 +/- 0.04 ml/dl x min/microg in the T3 and placebo study, respectively (P = 0.03). T3 infusion had no effect on the FBF response to sodium nitroprusside. T3 potentiated the vasoconstrictor response to norepinephrine (P = 0.006 for the interaction). Also, the slopes of the dose-response curves were affected by T3 (1.95 +/- 0.77 and 3.83 +/- 0.35 ml/dl x min/mg in the placebo and T3 study, respectively; P < 0.05). The increase in basal FBF induced by T3 was inhibited by NG-monomethyl-L-arginine. CONCLUSIONS: T3 exerts direct and acute effects on the resistance vessels by enhancing endothelial function and norepinephrine-induced vasoconstriction. The data may help clarify the vascular impact of the low T3 syndrome and point to potential therapeutic strategies.

Urinary gelatinase activities (matrix metalloproteinases 2 and 9) in human bladder tumors.

The ability to degrade type IV collagen, the major component of the basement membrane, is unique to gelatinases A and B. These two matrix metalloproteinases (MMPs) are most often linked to the malignant phenotype of tumor cells, and their expression is elevated in several cases of human tumor aggressiveness and overall survival. By gelatin zymography, we verified MMP activity in the urine of patients with bladder cancer. Of these patients, 10 had well-, 8 had moderately and 7 had poorly differentiated bladder cancer. The urine of healthy volunteers with no evidence of disease was used for controls. Zymography showed five dominant gelatinolytic bands of 240, 220, 130, 92 and 72 kDa in tumor samples, whereas only traces of MMP were detected in the urine of healthy subjects. The majority of cancerous urine samples showed MMP-9 lytic activity but only a few contained MMP-2. Moreover, MMP-9 content is enhanced in the urine from patients with high-grade and advanced-stage bladder tumors. Finally, we determined the urinary levels of urinary bladder cancer (UBC), tissue polypeptide-specific antigen (TPS) and protein 22 of nuclear matrix (NMP22). The levels of TPS and NMP-22 were higher in G3 bladder cancer than in G1 and G2 neoplasias. The urinary values of these two biomarkers correlated with the increase in MMP-9 lytic activity in high-grade and advanced-stage bladder cancer.

A mouse model demonstrates a multigenic origin of congenital hypothyroidism.

Congenital hypothyroidism with thyroid dysgenesis (TD) is a frequent human condition characterized by elevated levels of TSH in response to reduced thyroid hormone levels. Congenital hypothyroidism is a genetically heterogeneous disease. In the majority of cases studied, no causative mutations have been identified and very often the disease does not show a Mendelian transmission. However, in approximately 5% of cases, it can be a consequence of mutations in genes encoding the TSH receptor or the transcription factors TITF1, FOXE1, or PAX8. We report here that in mouse models, the combination of partial deficiencies in the Titf1 and Pax8 genes results in an overt TD phenotype that is absent in either of the singly deficient, heterozygous mice. The disease is characterized by a small thyroid gland, elevated levels of TSH, reduced thyroglobulin biosynthesis, and high occurrence of hemiagenesis. The observed phenotype is strain specific, and the pattern of transmission indicates that at least two other genes, in addition to Titf1 and Pax8, are necessary to generate the condition. These results show that TD can be of multigenic origin in mice and strongly suggest that a similar pathogenic mechanism may be observed in humans.

Analysis of glycoproteins in human colon cancers, normal tissues and in human colon carcinoma cells reactive with monoclonal antibody NCL-19-9.

The expression of sialyl Lewis(a) antigen, also known as Ca 19-9, in colon cancer, normal tissues and LS174T human colon carcinoma cells were studied. In colon adenocarcinoma and cell plasma membranes this antigen is expressed on various glycoproteins with different molecular weights ranging in size from over 200 kDa to about 100 kDa. In addition, there is very low expression in peritumoral tissues. In cytosol and culture medium this epitope is carried by a single complex-glycoprotein with a very high molecular weight resembling a mucin. In cells the rise in cAMP levels elevate the synthesis and release of the carbohydrate antigen 19-9; whereas the treatment with 1,9-dideoxyforskolin, a diterpene, which does not activate adenylate cyclase, has no effect on content of the antigen. These results suggest that cAMP is involved on the expression of glycoprotein-associated sialyl Lewis(a) antigen in LS174T cells.

Matrix metalloproteinase-2 and matrix metalloproteinase-9 type IV collagenases in serum of patients with pleural effusions.

Matrix metalloproteinases (MMPs) are proteolytic enzymes that are essentially involved in the turnover of the extracellular matrix (ECM). Recently, several MMPs were implicated in creating an environment that supports the initiation and maintenance of tumor growth. We verified MMP activity in the sera of patients with pleural effusions by gelatin zymography. Of these patients, 22 had malignant pleural effusion, consisting of 11 breast carcinomas and 11 lung carcinomas (7 squamous cell carcinomas and 4 adenocarcinomas), and 8 patients had benign effusions. The sera of 25 healthy subjects were used as controls. Zymography analysis showed three major gelatinolytic bands of 72, 92 and 220 kDa. The MMP-9/MMP-2 ratio was enhanced in cancer patients compared with benign diseases and healthy individuals. Furthermore, we determined the circulating levels of carcinoembryonic antigen (CEA), carbohydrate antigen 19-9 (Ca 19-9), carbohydrate antigen 15-3 (Ca 15-3). The serum levels of Ca 15-3 were above the cut-off levels in several cancer patients. No correlation between gelatinolytic activity and high tumoral marker values was found.

Rhes is involved in striatal function.

The development and the function of central nervous system depend on thyroid hormones. In humans, the lack of thyroid hormones causes cretinism, a syndrome of severe mental deficiency. It is assumed that thyroid hormones affect the normal development and function of the brain by activating or suppressing target gene expression because several genes expressed in the brain have been shown to be under thyroid hormone control. Among these, the Rhes gene, encoding a small GTP-binding protein, is predominantly expressed in the striatal region of the brain. To clarify the role of Rhes in vivo, we disrupted the Rhes gene by homologous recombination in embryonic stem cells and generated mice homozygous for the Rhes null mutation (Rhes(-/-)). Rhes(-/-) mice were viable but weighed less than wild-type mice. Furthermore, they showed behavioral abnormalities, displaying a gender-dependent increase in anxiety levels and a clear motor coordination deficit but no learning or memory impairment. These results suggest that Rhes disruption affects selected behavioral competencies.

New approaches in the diagnostic procedure of malignant pleural effusions.

The content of carcinoembryonic antigen (CEA), carbohydrate antigen 19-9 (Ca 19-9), carbohydrate antigen 15-3 (Ca 15-3) and the expression of LewisY related carbohydrate antigens in benign and malignant pleural effusion were determined. These included 35 malignant pleural effusions: 13 breast cancers, 12 lung cancers (6 squamous cell carcinomas, 5 adenocarcinomas and 1 microcytoma), 2 mesotheliomas, 1 epithelioma, 1 kidney cancer, 1 hepatocarcinoma, 1 colon carcinoma, 3 lymphomas, 1 osteosarcoma and 9 benign pleural effusions. We showed that pleural fluid content of CEA, Ca 19-9 and Ca 15-3 were higher in malignant than in benign effusions. However CEA levels in squamous lung cancers were very high in both serum and pleural fluids whereas its levels were only slightly above the cut-off in breast cancers and in lung adenocarcinomas. Serum and pleural fluid Ca 15-3 values were higher in breast and in lung cancers with the highest values in the patients with breast cancer. Furthermore, the LewisY related carbohydrate antigens, evaluated by the reactivity of the cell extracts to MAb B3, were expressed only in breast cancers. These data suggest that pleural fluid content of CEA, and Ca 15-3 associated with the immunoblotting of cell extracts with MAb B3 appear to be very useful to improve the diagnosis of malignant pleural effusions.

Role of cytokeratins, nuclear matrix proteins, Lewis antigen and epidermal growth factor receptor in human bladder tumors.

The content of urinary bladder cancer antigen (UBC), tissue polypeptide specific antigen (TPS), nuclear matrix protein 22 (NMP22), and the expression of LewisY carbohydrate antigens and of epidermal growth factor receptor (EGF-R) in bladder tumor tissues were determined. These included 14 well, 6 moderately and 11 poorly differentiated bladder cancers. Cytosol UBC and TPS were higher in the well and in the moderately differentiated bladder tumors than in the poorly differentiated bladder cancers; whereas cytosol NMP22 was higher in the poorly differentiated bladder cancers than in the well and in the moderately differentiated bladder tumors. The Lewis related carbohydrate antigens, evaluated by the reactivity of the tissues to monoclonal antibody B3, were highly expressed in poorly differentiated tumors. The EGF-R was strongly expressed in a large number of poorly differentiated bladder tumors. These data suggest that the determination of cytosol NMP22 and the immunoblotting with B3 and EGF-R antibodies might be useful to obtain more information on the differentiation of bladder tumors.

[Expression of Lewis carbohydrate antigen in human tumors]. / Espressione dell'antigene carboidratico Lewis nei tumori umani.

In this study we show, by immunoblotting, that Mab B3, a newly isolated monoclonal antibody, reacts with a variety of glycoproteins with different molecular weights expressed in gastric, pancreatic and colorectal cancers. The reactivity pattern differed in cancers arising in different tissues, though no correlation was observed with the histopathological characteristics of the lesions analysed. MAb B3 does not react with liver, brain or kidney cancers and has a limited reactivity with lung cancers but reacts very strongly with metastatic lesions. Because of the limited reactivity of this antibody with normal tissues, MAb B3 genetically fused with toxin in the form of a recombinant immunotoxin may be useful in treating certain kinds of cancer such as metastatic lesions. However, until current trials are completed, we will not know whether this immunotoxin will be helpful in cancer treatment.

Cyclic AMP-dependent secretion of Ca 19-9 by LS174T human colon carcinoma cells.

Prolonged increase of cyclic adenosine-monophosphate (cAMP) level in culture medium of a human colon cancer cell (LS174T) inhibits cellular growth and stimulates Ca 19-9 expression. The raise in cAMP level was produced by dibutyryl cyclic AMP (DBcAMP) or by forskolin an agent acting at the level of cAMP generation. Both these agents in a range of concentration between 10(-3)-10(-5) M have an inhibitory effect on the growth which is dose and time dependent. The inhibition was reversible as demonstrated by complete restoration of cell growth soon after the withdrawal of the substances from the culture medium. When cAMP levels in culture medium was raised, an increase in Ca 19-9 expression was observed and it appears that cyclic nucleotides have at least two effects: the first to cause rapid release of already synthesized Ca 19-9 and second to stimulate new antigen synthesis. The findings of the present study demonstrated that LS174T cells are unable to proliferate upon sustained accumulation of intracellular cyclic AMP suggesting the use of strategies able to increase cAMP levels for therapy of colon cancer. Furthermore, the finding that cAMP may also be a regulator of Ca 19-9 synthesis and release indicates the utility of cell line LS174T as a model for studies on the mechanism of synthesis and secretion of specific tumoral markers in colon cancer.