Quality of life compared during pharmacological treatments and clinical monitoring for non-localized prostate cancer: a randomized controlled trial.

OBJECTIVES: To investigate the effects of different management strategies for non-localized prostate cancer on men's quality of life and cognitive functioning. PATIENTS, SUBJECTS AND METHODS: Men with prostate cancer were randomly assigned to one of four treatment arms: leuprorelin, goserelin, cyproterone acetate (CPA), or close clinical monitoring. In a repeated-measures design, men were assessed before treatment (baseline) and after 6 and 12 months of treatment. A community comparison group of men of the same age with no prostate cancer participated for the same length of time. The men were recruited from public and private urology departments from university teaching hospitals. All those with prostate cancer who were eligible for hormonal therapy had no symptoms requiring immediate therapy. In all, 82 patients were randomized and 62 completed the 1-year study, and of the 20 community participants, 15 completed the study. The main outcome measures were obtained from questionnaires on emotional distress, existential satisfaction, physical function and symptoms, social and role function, subjective cognitive function, and sexual function, combined with standard neuropsychological tests of memory, attention, and executive functions. RESULTS: Sexual dysfunction increased for patients on androgen-suppressing therapies, and emotional distress increased in those assigned to CPA or close clinical monitoring. Compared with before treatment there was evidence of an adverse effect of leuprorelin, goserelin, and CPA on cognitive function. CONCLUSIONS: In deciding the timing of androgen suppression therapy for prostate cancer, consideration should be given to potential adverse effects on quality of life and cognitive function.

The use of isotope-coded affinity tags (ICAT) to study organelle proteomes in Arabidopsis thaliana.

Organelle proteomics is the analysis of the protein contents of a subcellular compartment. Proteins identified in subcellular proteomic studies can only be assigned to an organelle if there are no contaminants present in the sample preparation. As a result, the majority of plant organelle proteomic studies have focused on the chloroplast and mitochondria, which can be isolated relatively easily. However, the isolation of components of the endomembrane system is far more difficult due to their similar sizes and densities. For this reason, quantitative proteomics methods are being developed to enable the assignment of proteins to a specific component of the endomembrane system without the need to obtain pure organelles.

Altered cognitive function in men treated for prostate cancer with luteinizing hormone-releasing hormone analogues and cyproterone acetate: a randomized controlled trial.

OBJECTIVE: To report the first systematic investigation of the cognitive effects of luteinizing hormone-releasing hormone (LHRH) analogues in male patients, as LHRH analogues have been associated with memory impairments in women using these drugs for gynaecological conditions. PATIENTS AND METHODS: Eighty-two men with extraprostatic prostate cancer were randomly assigned to receive either continuous leuprorelin, goserelin (both LHRH analogues), cyproterone acetate (a steroidal antiandrogen) or close clinical monitoring. These patients underwent cognitive assessments at baseline and before starting treatment (77), and then 6 months later (65). RESULTS: Compared with the baseline assessments, men receiving androgen suppression monotherapy performed worse in two of 12 tests of attention and memory; 24 of 50 men randomized to active treatment and assessed 6 months later had a clinically significant decline in one or more cognitive tests but not one patient randomized to close monitoring showed a decline in any test performance. CONCLUSION: Pharmacological androgen suppression monotherapy for prostate cancer may be associated with impaired memory, attention and executive functions.

Identification of collagen fibril fusion during vertebrate tendon morphogenesis. The process relies on unipolar fibrils and is regulated by collagen-proteoglycan interaction.

The synthesis of an extracellular matrix containing long (approximately mm in length) collagen fibrils is fundamental to the normal morphogenesis of animal tissues. In this study we have direct evidence that fibroblasts synthesise transient early fibril intermediates (approximately 1 micrometer in length) that interact by tip-to-tip fusion to generate long fibrils seen in older tissues. Examination of early collagen fibrils from tendon showed that two types of early fibrils occur: unipolar fibrils (with carboxyl (C) and amino (N) ends) and bipolar fibrils (with two N-ends). End-to-end fusion requires the C-end of a unipolar fibril. Proteoglycans coated the shafts of the fibrils but not the tips. In the absence of proteoglycans the fibrils aggregated by side-to-side interactions. Therefore, proteoglycans promote tip-to-tip fusion and inhibit side-to-side fusion. This distribution of proteoglycan along the fibril required co-assembly of collagen and proteoglycan prior to fibril assembly. The study showed that collagen fibrillogenesis is a hierarchical process that depends on the unique structure of unipolar fibrils and a novel function of proteoglycans.

Surface located procollagen N-propeptides on dermatosparactic collagen fibrils are not cleaved by procollagen N-proteinase and do not inhibit binding of decorin to the fibril surface.

Dermatosparaxis is a recessive disorder of animals (including man) which is caused by mutations in the gene for the enzyme procollagen N-proteinase and is characterised by extreme skin fragility. Partial loss of enzyme activity results in accumulation of pNcollagen (collagen with N-propeptides) and abnormal collagen fibrils in the fragile skin. How the N-propeptides persist in the tissue and how abnormal fibril morphology results in fragile skin is poorly understood. Using biochemical and quantitative mass mapping electron microscopy we showed that the collagen fibrils in the skin of a dermatosparactic calf contained 57% type I pNcollagen and 43% type I collagen and the fibrils were irregularly arranged in bundles and hieroglyphic in cross-section. Image analysis of the fibril cross-sections suggested that the deviation from circularity of dermatosparactic fibrils was caused by N-propeptides of pNcollagen being located at the fibril surface. Comparison of experimental and theoretical axial mass distributions of the fibrils showed that the N-propeptides were located to the overlap zone of the fibril D-period (where D=67 nm, the characteristic axial periodicity of collagen fibrils). Treatment of the dermatosparactic fibrils with N-proteinase did not remove the N-propeptides from the fibrils, although the N-propeptides were efficiently removed by trypsin and chymotrypsin. However, the N-propeptides were efficiently cleaved by the N-proteinase when the pNcollagen molecules were extracted from the fibrils. These results are consistent with close packing of N-propeptides at the fibril surface which prevented cleavage by the N-proteinase. Long-range axial mass determination along the fibril length showed gross non-uniformity with multiple mass bulges. Of note is the skin fragility in dermatosparaxis, and also the appearance of mass bulges along the fibril long axis symptomatic of the fragile skin of mice which lack decorin. Western blot analysis showed that the dermatosparactic fibrils bound elevated levels of the proteoglycan, compared with normal skin fibrils. The results showed that N-propeptides can distort the morphology of fibrils, that they do not inhibit binding of gap-associated macromolecules (such as decorin) and that the normal mechanical properties of skin are strongly dependent on the close association of near-cylindrical fibrils, thereby enabling maximal fibril-fibril interactions.

Radical prostatectomy: is complete resection of the seminal vesicles really necessary?

PURPOSE: We determined the frequency of prostate cancer extension into the distal 1 cm. of seminal vesicles, and reconsidered whether complete excision of the seminal vesicles during radical prostatectomy is always necessary. MATERIAL AND METHODS: After en bloc removal with the specimen in 71 consecutive radical prostatectomies, the distal 1 cm. of each seminal vesicle was transected and separately analyzed for tumor involvement. RESULTS: Mean patient age was 61.8 years (range 40 to 72). Preoperative prostate specific antigen (PSA) ranged from 0.8 to 37 ng./dl. (median 7.3), and 18 patients had a PSA of 10 or more. Clinical stages were T1b in 1 case T1c in 37, T2a in 12, T2b in 10, T2c in 6 and T3a in 1. Preoperative Gleason sums ranged from 4 to 8 (median 6) with 21 patients (30%) having a sum of 7 or more. Of 71 patients 12 (17%) and seminal vesicle invasion (5 bilaterally). In no case did tumor extend into the distal 1 cm. of the seminal vesicle. PSA at diagnosis ranged from 4.2 to 30 ng./dl., with 4 of 12 patients having a PSA of 10 or more. Preoperative clinical stages were T1c in 5 cases, T2a in 3, T2b in 2 and T2c in 2. Five of the 12 patients (42%) had positive surgical margins and 11 (92%) had a postoperative Gleason sum of 7 or more. CONCLUSIONS: In 71 consecutive patients undergoing radical prostatectomy no tumor was found in the distal 1 cm. of the seminal vesicles, including 12 with seminal vesicle invasion. We continue to advocate complete excision of the seminal vesicles during radical prostatectomy. However, if dissection is difficult and a small fragment is left behind, the prognosis is unlikely to be altered.

Enzymic control of collagen fibril shape.

The shape of collagen fibrils growing in vitro in a cell-free enzyme/substrate system is shown to be dependent on the enzyme/substrate (E/S) ratio. Long fibrils with tapered ends were generated by exposing pCcollagen (procollagen from which the N-propeptides had been removed) to procollagen C-proteinase (which acts by cleaving the C-propeptides from the pCcollagen, converting it to insoluble fibril-forming collagen). Tip shape profiles, established quantitatively by scanning transmission electron microscopy, depended critically on the C-proteinase/pCcollagen ratio. The finest tips occurred at low ratios, the coarsest at high ratios. All fibrils had molecules oriented with amino termini closest to the pointed ends, i.e. N,N-bipolar fibrils in which molecules change orientation abruptly at one location along the fibril. Fibrils had maximal diameter at this molecular switch region. Shape asymmetric fibrils occurred at low E/S ratios, near-shape symmetric fibrils occurred at high ratios. Fibrils generated at low E/S ratios bore the closest resemblance to those formed in vivo except that the central shaft regions of fibrils formed in vitro showed no tendency to be limited to a uniform diameter.

Positive surgical margins with radical prostatectomy: detailed pathological analysis and prognosis.

OBJECTIVES: To examine the extent and location of positive surgical margins and their influence on progression. METHODS: Two hundred fifteen consecutive radical prostatectomy specimens, using 2 to 3-mm step-sections, were reviewed. Particular attention was paid to the location and extent of positive margins. Seventy-three patients (34%) with one or more positive margins were subjected to further detailed analysis. Progression was defined as a serum prostate-specific antigen level greater than 0.1 ng/mL and rising. The mean follow-up period was 23.2 months; median 24 months (range 3 to 40). RESULTS: Margin-positive patients had a significantly higher biopsy tumor grape (P = 0.05) than did margin-negative patients. Capsular preforation was present in 75%, seminal vesicle invasion in 33%, and nodal metastases in 10% of margin-positive patients; in contrast, these tumor characteristics were present in 47%, 8%, and 1% of margin-negative patients, respectively. The extent of involvement of linked margins was focal in 22% and extensive in 66%. An equivocal margin identified as surgical incision into the specimen (due to hemostatic staples, surgical dissection, or retraction) was present in 12%. Seventy-one percent of patients had a positive margin at only one location. Of all 99 positive-margin locations, 40% were apical, 10% anterior, 8% bladder neck, 16% posterolateral, and 25% posterior. Thirty-four percent of margin-positive and 7% of the margin-negative patients demonstrated biochemical progression. Of the 36 patients with a positive margin as their only major risk factor for progression (seminal vesicle and lymph node negative, Gleason score less than 8), 25% have progressed. Progression occurred in 2 of 9 patients with an equivocal positive margin, and 5 of 16 with a single focal-positive margin. A multivariate analysis of margin-positive patients identified tumor volume and grade as the most significant predictors, with the location and extent of the positive margin not significant. CONCLUSIONS: Although more frequent at the prostatic apex, tumor at the inked margin at any location is a risk factor for postoperative biochemical progression.

Neoadjuvant hormonal treatment before radical prostatectomy.

Randomized trials evaluating the use of androgen deprivation therapy (ADT) before radical prostatectomy have consistently shown significant decreases in prostate volume and in serum prostate-specific antigen (PSA) levels. All but one study have reported that hormonal pretreatment significantly reduces the incidence of positive surgical margins. However, androgen deprivation does not significantly influence seminal vesicle extension or lymph node involvement nor does it affect tumor grade. The fibrotic reactions that sometimes result from hormonal pretreatment can increase the difficulty of surgery but this has not been associated with a higher incidence of perioperative or postoperative complications. In closely monitored clinical trials, bothersome side effects of neoadjuvant hormonal therapy have been limited. Issues that remain to be addressed are the optimal duration of neoadjuvant treatment and whether the benefits of such therapy will translate into improved disease-free survival. At this time, patients with clinical stage T2b disease, PSA elevations greater than 10 to 20 ng/mL, and a high Gleason grade may be considered candidates for neoadjuvant hormonal treatment before surgery.

Identification of integrin alpha 2 beta 1 as cell surface receptor for the carboxyl-terminal propeptide of type I procollagen.

The carboxyl-terminal propeptide of procollagen type I (CPP-I) plays a key role in the regulation of collagen fibrillogenesis. In addition, it has been reported that, after cleavage from procollagen, CPP-I exerts feedback control of collagen biosynthesis. To further elucidate the mechanisms involved in each of these processes, we have investigated the nature of cell surface receptors for CPP-I. CPP-I affinity chromatography, using detergent extracts of iodinated HT1080 cells and EDTA elution, resulted in the isolation of two polypeptides of molecular mass 160 and 110 kDa. Since the migratory behavior of these polypeptides under nonreducing and reducing conditions was characteristic of a subset of integrin receptors, their reactivity with anti-integrin monoclonal antibodies was tested. Antibodies directed against the alpha 2 and beta 1 subunits specifically immunoprecipitated both CPP-I-binding polypeptides, indicating that the CPP-I receptor is the integrin alpha 2 beta 1. CPP-I was found to support the attachment and spreading of HT1080 cells, demonstrating that it can function as an adhesion protein. Two other approaches supported the identification of alpha 2 beta 1 as the CPP-I receptor. First, anti-functional anti-integrin monoclonal antibodies directed against the alpha 2 and beta 1 subunits completely abrogated the adhesive activity of CPP-I and, second, highly purified CPP-I bound specifically to alpha 2 beta 1-containing integrin preparations in a solid-phase receptor-ligand binding assay. These findings have important implications for the function of fibrillar collagen carboxyl-terminal propeptides and for the role played by integrins in the regulation of cellular phenotype.

Extracorporeal shock wave lithotripsy for ureteric calculi with the Dornier MFL5000 lithotriptor at a multi-user centre.

We have reviewed 241 consecutive cases of ureteric calculi managed with the Dornier MFL5000 lithotriptor. The patients were individually managed by 24 visiting urologists between October 1990 and September 1991. There were 153 cases of upper ureteric calculi, of which 59 were successfully manipulated back to the renal pelvis prior to treatment, and 94 were treated in situ. There were 27 mid-ureteric and 61 lower ureteric cases; in 51% of stones treated in situ ureteric stents were placed prior to extracorporeal shock wave lithotripsy. All stones were radio-opaque and localised with on-line fluoroscopy. The outcome of treatment was assessed at 3 months, with failure defined as residual calculi > or = 4 mm. The follow-up rate was 89%. The overall fragmentation rate after a single treatment was 72%, which increased to 81% with re-treatment. Of the upper ureteric calculi, manipulated stones had a significantly higher fragmentation rate (85%) compared with in situ stones (63%). The fragmentation rate was 89% for mid-ureteric stones and 71% for lower ureteric stones. Factors that significantly influenced fragmentation were retrograde manipulation of stone, number of impulses delivered and age of the patient. Stone size, the presence of a ureteric stent and mode of anaesthesia did not significantly influence fragmentation. These results suggest the Dornier MFL5000 lithotriptor is effective for the management of ureteric calculi in a multi-user setting.

Ehlers-Danlos syndrome type VIIB. Morphology of type I collagen fibrils formed in vivo and in vitro is determined by the conformation of the retained N-propeptide.

Previously we showed that fibrils generated from collagen and pNcollagen-ex6 from fibroblasts of an individual with Ehlers-Danlos syndrome (EDS) type VIIB were hieroglyphic in cross-section and all N-propeptides were located at the fibril surface. Hieroglyphs were resolved to near-cylindrical fibrils (that were similar in appearance to the fibrils seen in the tissues of individuals with EDS type VIIB) by treatment with N-proteinase which cleaved the pN alpha 1(I) chains but not the pN alpha 2(I)-ex6 chains (Watson, R. B., Wallis, G. A., Holmes, D. F., Viljoen, D., Byers, P. H., and Kadler, K. E. (1992) J. Biol. Chem. 267, 9093-9100). Here, quantitative scanning transmission electron microscopy (STEM) showed that N-propeptides in hieroglyphs were in a "bent-back" conformation and thus located exclusively in the overlap zone of the fibril D-period (D = 67 nm). In contrast, STEM of fibrils from the dermis of an individual with EDS type VIIB showed that partially cleaved N-propeptides (in which cleaved pN alpha 1(I) remained in noncovalent association with pN alpha 2(I)-ex6 chains) were distributed equally between the gap and overlap zones of the fibrils. Comparison of experimental data with theoretical mass distributions of the fibril based on amino acid sequence data gave a consistent value of 33 nm for the total axial extent for the N-propeptides in hieroglyphic and tissue fibrils irrespective of the location of N-propeptides to the gap or overlap zone. These data exclude the possibility that N-propeptides adopt a random configuration, but rather, that they locate to specific sites in the gap and overlap zones. The results demonstrated that cleavage of pN alpha 1(I) chains in vivo releases the N-propeptides from the constraints of the bent-back conformation. Co-distribution of partially cleaved N-propeptides between gap and overlap zones allows a higher surface packing density of N-propeptides and explains how circularity of large diameter fibrils can be achieved despite the retention of N-propeptides in tissues of individuals with EDS type VIIB.

Ehlers Danlos syndrome type VIIB. Incomplete cleavage of abnormal type I procollagen by N-proteinase in vitro results in the formation of copolymers of collagen and partially cleaved pNcollagen that are near circular in cross-section.

We have shown that a child with Ehlers Danlos syndrome (EDS) type VII has a G to A transition at the first nucleotide of intron 6 in one of her COL1A2 alleles. Half of the cDNA clones prepared from the proband's pro alpha 2(I) mRNA lacked exon 6. The type I procollagen secreted by the proband's dermal fibroblasts in culture was purified, and collagen fibrils were generated in vitro by cleavage of the procollagen with the procollagen N- and C-proteinases. Incubation of the procollagen with N-proteinase resulted in a 1:1 mixture of pCcollagen and uncleaved procollagen. Incubation of this mixture with C-proteinase generated collagen and abnormal pNcollagen (pNcollagen-ex6) that readily copolymerized into fibrils. By electron microscopy these fibrils resembled the hieroglyphic fibrils seen in the N-proteinase-deficient skin of dermatosparactic animals and humans and were distinct from the near circular cross-section fibrils seen in the tissues of individuals with EDS type VII. Further incubation of the hieroglyphic fibrils with N-proteinase resulted in partial cleavage of the pNcollagen-ex6 in which the abnormal pN alpha 2(I) chains remained intact. These fibrils were not hieroglyphic but were near circular in cross-section. Fibrils formed from collagen and pNcollagen-ex6 that had been partially cleaved with elevated amounts of N-proteinase prior to fibril formation were also near circular in cross-section. The results are consistent with a model of collagen fibril formation in which the intact N-propeptides are located exclusively at the surface of the hieroglyphic fibrils. Partial cleavage of the pNcollagen-ex6 by N-proteinase allows the N-propeptides to be incorporated within the body of the fibrils. The model provides an explanation for the morphology and molecular composition of collagen fibrils in the tissues of patients with EDS type VII.

gamma IFN receptor expression in haemic malignancies.

gamma Interferon receptor (gamma IFNR) expression was examined by Scatchard analysis in 49 patients with various haemic malignancies. In chronic lymphocytic leukaemia (CLL) (20 cases) expression was variable but generally low (0-1600 receptors/cell) and was not related to clinical stage. In hairy-cell leukaemia (HCL) (n = 6), prolymphocytic leukaemia (PLL) (n = 1) and acute lymphoblastic leukaemia (ALL) (n = 4) expression was again variable but was significantly higher than in CLL (500-4500 receptors/cell). In the myeloid proliferations (n = 18) receptor numbers were significantly higher than in the lymphoproliferative disorders as a whole (1000-30,000) and the highest level of expression was observed in primitive monocytoid proliferations.

Cigarette smoking behavioral distinctions between experimental nonadopters and adopters in children and adolescents--a consideration of transitional smoking experience: the Bogalusa Heart Study.

A cigarette-smoking questionnaire to examine behavior, attitudes, and beliefs related to cigarette use was administered to children, ages 8-17, in a biracial community. Children who experimented with cigarettes but did not adopt the habit (experimental nonadopters) and children who continued to smoke (adopters) were identified and characterized. Follow-up behavior was examined 2 years later. Adopters were more likely to have smokers as friends and family members, more likely to have purchased their first cigarettes, more likely to believe smoking to be pleasurable for themselves and others, and less likely to consider smoking harmful. Adopters who maintained smoking behavior 2 years later had, during the initial survey, reported having more friends who also smoked and were more likely to believe smoking to be enjoyable. Experimental nonadopters were more likely to try the first cigarette alone, reported having fewer friends and family members who smoked, and believed greater health risks to be associated with cigarette use. Experimental nonadopters who maintained nonsmoking behavior 2 years later, especially in the older cohort, exhibited higher agreement with the negative consequences of cigarette smoking (health beliefs) and theories concerning smoking behavior of others.