Evaluating the efficacy of acupuncture in defined aspects of stroke recovery: a randomised, placebo controlled single blind study.

OBJECTIVE: To investigate the efficacy of acupuncture on stroke recovery compared to an inert placebo. DESIGN: Placebo-controlled, randomised, clinical trial. SETTING: Post-stroke rehabilitation wards in five NHS hospitals in the UK. SUBJECTS: Patients between 4 and 10 days after their first stroke. INTERVENTIONS AND OUTCOME MEASURES: The patients received 12 acupuncture or placebo treatments over four weeks. Acupuncture with electrical stimulation was compared with mock TENS, and assessments continued for 12 months after entry. Primary outcome was the Barthel Index (BI). Secondary outcomes were muscle power, Motricity Index (MI), mood, Nottingham Health Profile (NHP) and treatment credibility. RESULTS: 92 patients completed data sets. Data were analysed using both t tests and a structural equation based on longitudinal analysis of both BI and MI, using generalised estimating equations with an exchangeable correlation structure. While both acupuncture and placebo (mock TENS) appeared to have had an equal effect on stroke recovery, there is no significant difference between the two interventions at 12 (p = 0.737, 95 % CI -2.00 to 2.81) and 52 weeks (p = 0.371, 95 % CI -3.48 to 1.32). An apparently accelerated improvement in the MI scores in the acupuncture group at 3 weeks (p = 0.009, 95 % CI 1.55 to 10.77) is interesting. CONCLUSIONS: Acupuncture did not demonstrate specific efficacy over placebo and both groups did as well as normally expected with this condition.

Androgen-independent cancer progression and bone metastasis in the LNCaP model of human prostate cancer.

Our laboratory has previously reported on the derivation of LNCaP cell sublines from LNCaP tumors maintained in castrated and intact athymic male mice. These LNCaP sublines differ from the parental line in tumorigenicity and androgen dependence. This paper demonstrates that one of these sublines acquired metastatic potential. When inoculated either s.c. or orthotopically, the C4-2 subline metastasized to the lymph node and bone with an incidence of 11-50%. Interestingly, the incidence of osseous metastasis was higher in castrated than in intact male hosts. We evaluated the chromosomal, immunohistochemical, and biochemical characteristics of the LNCaP sublines derived from C4-2 tumors that metastasized to the lymph node and bone. Cytogenetic analysis showed that all sublines were human and shared common marker chromosomes with the parental LNCaP cells. This experimental human prostate cancer model may permit, for the first time, the study of the molecular mechanisms underlying human prostate cancer metastasis.

Involvement of chromosome 7 in primary lung tumor and nonmalignant normal lung tissue.

By using the newly developed adhesive tumor cell culture system, we analyzed the chromosomal constitutions of primary lung tumor and nonmalignant normal lung tissue from 10 previously untreated patients with non-small cell lung cancer. Chromosomal analyses were successfully carried out in banded chromosome preparations from 10 tumor and 8 normal lung tissue samples. All analyzed tumor and normal lung tissue samples had a predominantly normal diploid chromosome number. However, there was at least one structural or numerical alteration in every tumor and lung tissue sample analyzed. Chromosomes 1, 3, 4, 6, 7, 8, 9, 12, 15, and 20 were more often involved in rearrangement. The most consistent finding was trisomy 7; 4 patients had trisomy 7 in both tumor and normal lung tissue, and another 2 had this anomaly in tumor tissue only. Of the 4 patients without trisomy 7, 2 had a homogeneously staining region in the short arm of chromosome 7 in tumor tissue. Phytohemagglutinin-stimulated peripheral blood lymphocytes from 7 patients, including 5 patients with trisomy 7 in tumor tissue, did not show trisomy 7. These cytogenetic data suggest that chromosome 7 may be associated with lung cancer development and that trisomy 7 may be the hallmark of premalignant changes, at least in a subgroup of patients with non-small cell lung cancer.

DNA repair and mutagen sensitivity in patients with triple primary cancers.

The purpose of this study was to measure DNA repair capacity and mutagen sensitivity in patients who have had three or more primary forms of cancer. It was hypothesized that, if abnormalities in DNA repair and mutagen sensitivity were cancer susceptibility factors, such findings would be seen with regularity in individuals with multiple primary cancers. DNA repair capacity was measured by determining repair of UV-irradiated plasmid DNA (pCMVCAT) transfected into peripheral blood lymphocytes. Results from 18 patients and a like number of age- and sex-matched controls demonstrated a significant difference in DNA repair capacity (P < 0.0001; odds ratio = 14). Mutagen sensitivity was measured by determining the mean number of chromatid breaks per cell after in vitro exposure to either bleomycin or 4-nitroquinoline-1-oxide. The difference in mean bleomycin- or 4-nitroquinoline-1-oxide-induced mutagen sensitivity between cases and controls was not statistically significant. Fourteen of the 18 patients had positive family histories of cancer; in 10, the history was compatible with cancer susceptibility syndromes. Although the numbers were small, there was no suggestion in this study that treatment or the presence of cancer was the cause of the DNA repair abnormalities encountered. These findings support the concept of diminished DNA repair capacity as an underlying feature in the development of a mutator phenotype.

Nonrandom chromosomal abnormalities in lymphocyte cultures of individuals with colorectal polyps and of asymptomatic relatives of patients with colorectal cancer or polyps.

We studied chromosomal alterations in the peripheral blood lymphocytes of 10 individuals with colorectal polyps and 10 asymptomatic first-degree relatives of patients with colon cancer or colorectal polyps. The analysis was performed on T-lymphocytes using short term blood cultures and on B-lymphocytes by establishing lymphoblastoid cell lines by Epstein-Barr virus transformation. Chromosomal changes were not common in T- and B-lymphocytes. Chromosomes 1 and 5 were most frequently involved in numerical or structural changes in the patients with polyps as well as in the asymptomatic relatives. These alterations were observed in either the T-lymphocytes or the B-lymphocytes but rarely in both, thus accentuating the importance of studying both the cultures concurrently. Chromosome 5, which is known to play an important role in the development of adenomatous polyps, was found to be involved in 6 (60%) of 10 patients with polyps and 4 (40%) of 10 asymptomatic relatives. These findings show that lymphocytic chromosomal analysis can aid in identifying individuals who are genetically susceptible and are at a higher risk of developing colorectal cancer. Because lymphocytic chromosomal analysis is relatively simple and inexpensive, we expect that it will be very useful in screening asymptomatic individuals who are at a higher risk due to inherited or environmental factors.

Genetic susceptibility to lung cancer as determined by lymphocytic chromosome analysis.

Chromosomal anomalies were analyzed in the lymphocyte cultures among 96 untreated lung cancer patients and 74 clinically normal comparison subjects. The analysis revealed that >15% of the lung cancer patients showed structural or numerical rearrangements in chromosomes 1,3,5,7,9,12,14, and 21. A case control comparison showed that these aberrations were significantly higher in chromosome 7 [odds ratio (OR) = 2.32; 95% confidence interval (CI), 1.14 and 4.82], chromosome 9 (OR = 2.61; 95% CI, 1.27 and 5.48), chromosome 12 (OR = 4.10; 95% CI, 1.40 and 14.54), and chromosome 21 (OR = 7.75; 95% CI, 1.73 and 70.80) of the patients than in the controls. However, only chromosome 9 (OR = 3.57; 95% CI, 1.33 and 9.46) and chromosome 21 (OR = 6.94; 95% CI, 3.15 and 9.98) retained significance after stratifying on smoking status. Among the lung cancer patients, the breakpoints cluster in specific regions of some of these chromosomes. These regions are 1p13-q21, 3q21-q13, 7p12-q12, 7q12-q12,7q22, 7q32, 9p13-q13, 12p13, 14q11, and 14q32. The distribution of lung cancer patients, according to histological types, showed that aberrations in chromosomes 1,7, and 9 dominated the scenario of chromosomal changes in non-small cell lung carcinomas. Thus, the data on lymphocytic chromosomal rearrangements in lung cancer patients not only indicate the importance of specific genetic changes in the etiology of lung cancer but also emphasizes the putative role of such analysis in determining primary genetic abnormalities in the large heterogeneous group of lung cancers.

Integrin-mediated entry into S phase of human gastric adenocarcinoma cells.

The integrins are a family of integral membrane receptors that participate in binding to various extracellular and cell surface proteins during adhesion, migration, and homing of normal and neoplastic cells. In this study, we characterized the involvement of integrins in mediating the growth of an adhesion-dependent gastric adenocarcinoma line, ST2. This line was distinguished and selected for study based on its inability to grow when suspended in soft agar or plated on poly(2-hydroxyethyl methacrylate)-coated dishes. ST2 cells arrested in G0/G1 of the cell cycle when deprived of adhesion to substrate. Using purified matrix components, collagen was found to be highly active in promoting beta 1 integrin-mediated cell attachment and spreading. Subsequent to spreading on collagen, the cells were released from G0/G1 block and progressed into S phase. Monoclonal antibodies to alpha 2 or beta 1 integrin blocked the reinduction of both cell spreading and entry into S phase. These studies suggest that during the metastatic process, integrin receptor interaction with the insoluble matrix may be an important step leading to proliferation of some tumors.

A comparison of methods for the detection of Candida antigens. Evaluation of a new latex reagent.

The sensitivity of various serological techniques for the detection of C. albicans cytoplasmic antigen (Ag) in buffer and serum diluents was compared, with special reference to variations of the ELISA method and a new microtitre latex particle agglutination (MLA) test. Of the assays evaluated, immunodiffusion, counterimmunoelectrophoresis and slide latex particle agglutination (SLA) were the least sensitive. ELISA tests were more sensitive (50-150 ng/ml Ag in buffer) although sensitivity decreased in serum or heat-inactivated serum (HIS) (250 ng/ml-4 micrograms/ml). The new MLA test had better sensitivity (16 ng/ml Ag in buffer) than any of the ELISA tests and was unaffected by the presence of serum or HIS (2.5 and 20 ng/ml Ag respectively). MLA seems worthy of further evaluation as an alternative to ELISA for use in antigen detection systems in general and for the serodiagnosis of systemic candidosis in particular.

Ag-NOR studies in a human lymphocyte culture: are variants localized to specific chromosomes?

The genomic activity and polymorphisms of nucleolus organizer regions (NORs) were studied in an individual with routinely used AgNO3 and Q-banding techniques. The results demonstrate that (a) the mean number of NORs per metaphase spread was 7.4, (b) chromosomes 14, 13, 15, and 21 showed variations of Ag-NOR in decreasing order, (c) chromosome 22 did not show a polymorphism in any cell examined, and (d) both chromosomes 14 showed silver staining in most cells. Unlike many earlier reports indicating that NOR variants were constitutional in each individual, our present case represented a mosaic pattern of polymorphism involving most of the D- and G-group chromosomes.

Shared cytogenetic abnormalities in lung-tumors and corresponding peripheral-blood lymphocytes.

We analyzed chromosomal alterations in primary lung tumours and peripheral blood lymphocytes (PBLs) from 10 lung cancer patients (nine with non-small cell lung carcinoma and one with small cell lung carcinoma) to determine whether there were shared chromosomal changes in the normal and diseased tissue of these cases. This study revealed that each paired sample had at least three chromosomes and two chromosomal regions in common for structural rearrangements. The chromosomes most frequently found structurally altered in paired analysis were 1 and 3 (60% each), 5 (50%), 6, 7, and 9 (40% each), and 14 (30%). Chromosome region 3p13-3p21 was structurally rearranged in both the normal and tumour tissues of three patients. Chromosome 3 was structurally rearranged in all ten tumours. The chromosome arms most commonly affected in the tumours were 3p (nine times), 9p and 5q (eight times each), Ip and 7q (six times each), 10q and 11q (five times each), 14q and 6q (four times each). The most frequently affected chromosomal regions in these tumours were (in decreasing order) 9p23-p24, 3p21-3p13, 5q11, 1p34, 7q22, and 11q13. Frequent polysomy of 7 and 12 and loss of D-group chromosomes were also observed in the tumours analyzed. Comparing the changes found only in tumours with those found in both PBLs and tumours was helpful in shedding some light on the probable sequence of genetic events leading to lung cancer. This investigation also offered compelling evidence that genomic instability at the chromosomal level in PBLs corresponds with the genetic changes observed in tumours indicating that PBL analysis can help identify the early chromosomal changes in lung cancer. PBL chromosomal analysis thus has a promising future in the genetic analysis of lung cancers.

Bcl-2 oncogene blocks differentiation and extends viability but does not immortalize normal human keratinocytes.

To ascertain whether the bcl-2 oncogene plays a role in the initial stages of skin carcinogenesis by preventing differentiation of epidermal keratinocytes, we transfected primary human keratinocytes with the human bcl-2 gene and then determined whether these transfectants escape high calcium- and serum-induced differentiation. We found that the bcl-2 oncogene blocked differentiation and extended the life span of human keratinocytes in culture by over 24 weeks compared with cells transfected with pZip-neo DNA, which only grew for 5 weeks in culture. Keratinocytes transfected with the bcl-2 oncogene exhibited apoptotic bodies and telomere-telomere association between chromosomes toward the end of their life-span. These results suggest that die bcl-2 oncogene may be necessary but not sufficient for the immortalization of human keratinocytes.

Identification of primary chromosome-abnormalities in a patient with endometrial carcinoma - analyses of tumor-biopsy and lymphocyte-cultures.

The development of human cancer is generally considered to be the result of genetic mutations that cause a progressively more malignant phenotype. We propose that such genetic changes can be observed in a small number of lymphocytic metaphase plates. We have identified a specific chromosome marker formation in a primary endometrial adenocarcinoma obtained from a 74-year-old woman. After observing an isochromosome for 1q in the tumor cells, we predicted that in her lymphocytes this particular chromosome must show susceptibility to breakage. After 6 months, when lymphocytes were available from this patient, 4.0% of her metaphases exhibited chromatid breaks in the pericentromeric region of one homolog of chromosome 1, thus confirming our prediction. Since then, the primary endometrial tumor cell line has been passaged through nude mice and has become highly metastatic. Examination of tumors obtained from different organ sites of these mice has revealed that the same altered homolog 1 underwent various types of chromosome and chromatid aberrations, thereby confirming the presence of instability in this particular chromosome in this particular cancer. A detailed karyotypic evolution from normal lymphocyte cultures --> primary endometrial tumor --> highly metastatic endometrial tumor was therefore possible to construct. Our results further support the idea that peripheral blood lymphocytes can be used as the tissue for studying genetics of cancer predisposition.

Correlation of non-random chromosomal aberrations in lymphocytes of prostate cancer patients with specific clinical parameters.

The purpose of this research was to correlate non-random chromosomal aberrations in the peripheral blood lymphocytes (PBLs) of prostate cancer patients with specific clinical parameters. Peripheral blood samples were analyzed from 59 informative prostate cancer patients. Non-random chromosomal alterations detected in the PBLs and their correlation with any specific clinical parameters were analyzed statistically. A comparison was made between specific chromosomal abnormalities in the patients having an early (<65 years) or late (> or =65 years) age at disease onset, low-grade (Gleason grade <7) or high-grade (Gleason grade > or =7) tumors, a low (<10 ng/ml) or high (> or =10 ng/ml) prostate-specific antigen (PSA) level, and androgen-sensitive or -insensitive disease. In examining the specific chromosomal breakpoints, the regions 1p13, 2q21, 3p21, 4q13, 5q31, 6p21, 7p15, 7p13, 7q32, 10p11, 10q26, 11p15, 11p11, 14q12, and 16q12 showed breaks in at least four cases. Chromosome 15 (P=0. 045) was significantly altered in patients having a PSA value greater than or equal to 10, while it (P=0.017) and chromosome 19 (P=0.036) were significantly altered in patients having a PSA value greater than or equal to 20. In addition, chromosomes 5 (P=0.032), 8 (P=0.020), 16 (P=0.009), and 20 (P=0.047) were significantly altered in patients having a Gleason grade greater than 7. Also, chromosomes 2 (P=0.020) and 3 (P=0.044) were significantly altered in patients who had early disease onset. Additionally, chromosome 10 (P=0.041) was significantly altered in patients having metastasis, and chromosomes 4 (P=0.006) and 7 (P=0.028) were significantly altered in patients having androgen-insensitive disease. In spite of the small subset of patients, chromosome 8 (p=0.003) was significantly altered in patients having small cell carcinoma of the prostate. From these results we conclude that non-random chromosomal aberrations present in PBLs of prostate cancer patients can be correlated with specific clinical parameters. These correlations can be used to identify a prostate cancer patient's risk response to therapy.

Rapid diagnosis of vaginal candidosis by latex particle agglutination.

Vaginal swabs from women who on clinical evidence were thought to have vaginal candidosis were examined for yeasts by conventional laboratory methods (microscopy and culture) and also assayed for Candida antigens using a rapid (3 min) slide latex particle agglutination tests. Results showed that a diagnosis of vaginal candidosis based on clinical criteria alone is unreliable: only half of the women were subsequently confirmed as having candidosis by microscopy and culture. The new slide latex particle agglutination test gave better results, with 100% specificity, 80% sensitivity, high predictive values (greater than or equal to 91%), and an overall diagnostic efficiency of 93%. From the results of this preliminary study, slide latex particle agglutination looks a promising, rapid alternative to conventional laboratory methods for confirming a clinical diagnosis of vaginal candidosis and has the considerable advantage that it can be conveniently used in a clinical setting.

Use of the Pastorex aspergillus antigen latex agglutination test for the diagnosis of invasive aspergillosis.

AIMS: To evaluate the Pastorex aspergillus antigen latex agglutination test for the diagnosis of invasive aspergillosis in patients undergoing liver or bone marrow transplantation. METHODS: Serum samples were taken at least twice weekly post-transplant and tested for Aspergillus antigen. Latex agglutination test results were compared with microbiological examination of respiratory, urine and bile specimens. Serum samples from liver transplant patients were also tested for antibodies to Aspergillus fumigatus by counter immunoelectrophoresis. RESULTS: Eight of the 91 patients studied developed invasive aspergillosis. Positive latex agglutination tests were obtained in eight of 187 (4.3%) serum samples from four of these eight patients. The other four patients with invasive aspergillosis gave consistently negative latex agglutination tests. A positive latex agglutination test was the first indication of invasive aspergillosis in two patients; these patients were already on amphotericin B. Positive latex agglutination tests were the only evidence of invasive aspergillosis in one patient who subsequently died of the infection. False positive latex agglutination tests were obtained in five of 83 (6%) patients with no evidence of invasive aspergillosis and misleading positive cultures seen in nine of 83 (10.8%). No antibodies were detected in three of four liver transplant patients with invasive aspergillosis. Conversely, antibodies were detected in 63 of 262 (24%) serum samples from 43 liver transplant patients with no evidence of invasive aspergillosis; one of these patients had an antibody titre of 1:2 on four separate occasions. CONCLUSIONS: The Pastorex aspergillus antigen latex agglutination test, when used alone, lacks sensitivity and specificity for the early diagnosis of invasive aspergillosis. A diagnosis was made in all patients with invasive aspergillosis when both culture and antigen tests were performed although using these criteria a false positive diagnosis would have been made in 13 of 83 (15.6%) patients. Microbiological and serial serological investigations for antigen should both be performed and the results considered in conjunction with radiological and clinical evidence.

Electric and chemical fusions for the production of monoclonal antibodies reacting with the in-vivo growth phase of Candida albicans.

To obtain monoclonal antibodies (MAbs) directed preferentially against the pathogenic phase of Candida albicans, mice were immunised with germ tubes of C. albicans serotype A, strain VW.32, killed by exposure to ultraviolet (UV) irradiation. Fusions were performed either by the standard chemical procedure with polyethylene glycol, or by electric discharge following linkage of the myeloma and lymphocyte cells with a Concanavalin A-mannoprotein bridge. The preliminary characteristics of one MAb obtained from each of these fusions are described. An IgM antibody (3B7) obtained from the chemical fusion reacted with a polysaccharide antigen that was heterogeneously distributed on both in-vitro and in-vivo forms of C. albicans. This MAb agglutinated different strains of C. albicans irrespective of their serotype. An IgG1 antibody (3G6) that had been obtained from the electric fusion was found to react in vitro with a proteinaceous antigen located only on the germ tubes of strain VW.32. However, MAb 3G6 displayed strong reactivity against all growth forms of C. albicans in vivo and reactivity extended to other strains.

Reaction of Candida albicans cells of different morphology index with monoclonal antibodies specific for the hyphal form.

Two monoclonal antibodies (MAbs), 3D9 with reported specificity for Candida albicans hyphae, and 3B7 with reported specificity for morphological forms of C. albicans found in vivo, were tested by indirect immunofluorescence with C. albicans cells that were grown in 12 different environments (four different culture media incubated at various temperatures) and whose cellular morphology was estimated in terms of morphology index (Mi). Both MAbs reacted strongly with cells with Mi greater than 3.0, i.e., with pseudohyphal and hyphal forms, but in Eagle's medium at 26 degrees C and in a modified Sabouraud's broth medium at 30 degrees C, some reactivity was also found with cells of lower Mi (i.e., yeast forms). Therefore, it was concluded that the hyphal phenotype and the epitopes reactive with the MAbs were co-expressed but that the epitopes could also be expressed independently of the hyphal phenotype. The results confirm the propensity of C. albicans for variation of its surface antigenic composition.

Trisomy 12 in Epstein-Barr virus-transformed lymphoblastoid cell lines of normal individuals and patients with nonhematologic malignancies.

Karyotypes of 36 lymphoblastoid cell lines established by Epstein-Barr virus (EBV) transformation of peripheral blood lymphocytes (PBL) of eight normal individuals and 28 patients with various nonhematologic malignancies were analyzed. In seven lines (19.4%), cells with trisomy 12 were noted, with clonality in two of these lines. In two of 11 metaphases with such trisomy, chromosome 12 was involved in structural rearrangements [t(8;12)(q12;p12) and t(12;12)(q11;q24)]. No cells with trisomy 12 were observed in phytohemagglutinin (PHA)-stimulated PBL cultures of these individuals. In 250 individuals (normal and with nonhematologic malignancies) examined in our laboratory in the last 5 years, extra copies of chromosome 12 in PHA-stimulated PBL cultures were observed in only five of 23,216 cells (0.02%). There were no cases of clonality in these samples. The frequency of an extra chromosome 12 was comparable to that of the other chromosomes except 21 and X, whose frequency of occurrence was 0.08% and 0.09%, respectively. These findings should be considered random events in PHA-stimulated PBL. On the contrary, in lymphoblastoid cell lines established by EBV transformation, trisomy of chromosome 12 was the most frequent numerical abnormality. It was observed in 64.7% of all cases with chromosome gains and therefore could not be considered a random occurrence. The specificity of this phenomenon for EBV transformation is supported by the results of cytogenetic analysis of eight lymphoblastoid cell lines established by an alternative procedure in our laboratory [1]. In 400 cells analyzed not a single cell with trisomy 12 was observed. We suggest that EBV transformation might either randomly induce formation of such cells in immortalized B-cell populations or show potentially blastomogenic cells or proneness to their formation in certain individuals who could be predisposed to develop lymphoproliferative diseases, especially chronic lymphocytic leukemia (CLL) in which trisomy of chromosome 12 is the most common alteration.

Sex chromosome abnormalities in lung cancer patients.

Chromosomal analyses of lymphocytes from lung cancer patients and normal subjects revealed that the X and the Y chromosomes have both structural and numerical abnormalities in higher frequency in patients compared to the controls. These abnormalities included chromatid/isochromatid breaks, translocations, ring formation, and selective nondisjunctions, resulting in multisomies of either the X or Y chromosomes. Possible significance of these genetic abnormalities are discussed in relation to lung cancer patients.

Aneuploidy index in blood: a potential marker for early onset, androgen response, and metastasis in human prostate cancer.

OBJECTIVES: To investigate whether the frequency of chromosome abnormalities in peripheral blood lymphocytes defined as the aneuploidy index in blood (AnIB) can be used as a clinical marker of early age onset, androgen response, and metastasis in human prostate cancer. METHODS: Peripheral blood samples were collected from 80 patients with prostate cancer, and chromosome preparations were made from 72-hour cultures after mitotic block. The AnIB of 59 informative cases was compared with several parameters, including age at disease onset, Gleason grade of tumor, clinical stage of tumor, metastasis, and prostate-specific antigen (PSA) level. RESULTS: Patients with AnIB levels greater than 3 had a significantly higher incidence of metastasis (P = 0.022), androgen-independent disease (P = 0.002), and early age at disease onset (age at diagnosis less than 65 years) (P = 0.002) compared with the patients with lower AnIB (less than 3) levels. In addition, patients with AnIB levels greater than 5 had higher PSA levels (greater than 20 ng/mL) (P = 0.029) than patients with AnIB levels less than 5. CONCLUSIONS: Chromosome abnormalities can be detected in the peripheral lymphocytes of patients with prostate cancer, and AnIB can be used as an early diagnostic and predictive marker for prostate cancer metastasis and androgen-independent disease.