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Biomedical subjects

R A Gardiner

Publications and source records attributed to R A Gardiner.

At least 19 recordsLinked to original sources
PubMed1999

Molecular detection of prostate cells in ejaculate and urethral washings in men with suspected prostate cancer.

PURPOSE: To determine whether prostatic cells were normally present in ejaculate and if the sensitivity and specificity of the detection of malignant prostate cells in ejaculate and urethral washings from men with suspected prostate cancer could be improved using the more sensitive molecular technique of reverse transcriptase-polymerase chain reaction (RT-PCR). MATERIALS AND METHODS: RT-PCR for prostate-specific antigen (PSA), prostate-specific membrane antigen (PSM) and Apoliprotein D (3 putative prostate-specific and/or cancer-specific markers) was performed on RNA extracts of ejaculate (80) and urethral washings (52) from 77 men with suspected prostate cancer and 12 young controls (<30 years of age) and urines from 5 men who had radical prostatectomies and 10 women. RESULTS: PSA, PSM and Apolipoprotein D expression was detected in ejaculates and urethral washings from both patient and control groups. No differences were observed in the results obtained for 58 men with suspected or 19 men with confirmed prostate cancer or the 18 vasectomized men within the patient group. Urines from the 5 men who had radical prostatectomies and 10 women were all negative for PSA, but PSM was detected in 2 female urines and in 3 radical prostatectomy samples. As few as 10 LNCaP prostate tumor cells could be detected by PSA RT-PCR when added to female urine. CONCLUSION: We have established a sensitive method of detecting prostatic cells in ejaculate and urethral washings and shown that PSA RT-PCR is a reliable indicator of prostate cells in these samples. However, RT-PCR for PSA, PSM and Apoliprotein D were not useful for discriminating malignant from non-malignant prostate cells.

Adult↗
PubMed1998

Evaluation of the potential of transferrin-adriamycin conjugates in the treatment of bladder cancer.

OBJECTIVE: To assess the ability of a transferrin-adriamycin conjugate (Tf-ADR) to target transferrin receptor (TfR)-positive cancer cells selectively and to overcome drug resistance in bladder cancer cell lines. MATERIALS AND METHODS: Two paired sets of cell lines were used: the first was Chinese hamster ovary (CHO) cells (TfR-negative TRVb cells, as a model for normal resting cells, and TRVb-1 cells which were transfected with human TfR), and the second was a pair of bladder cancer cell lines (ADR-sensitive MGH-U1 cells and ADR-resistant MGH-U1R cells). Cell survival curves were determined after treatment with ADR, Tf and Tf-ADR. RESULTS: MGH-U1, TRVb and TRVb-1 cells required similar concentrations of ADR and Tf-ADR for 50% inhibition of growth; MGH-U1R cells were resistant to both ADR and TF-ADR. CONCLUSION: Tf-ADR did not prevent toxicity to the TfR-negative cells nor did it overcome the resistance of the ADR-resistant cells. These results imply that Tf-ADR does not provide a better cytotoxic drug delivery system for the treatment of bladder cancer.

Animals↗
PubMed1998

Experimental pyeloureterectomy and calyceal neck transection with autotransplantation and bladder advancement.

OBJECTIVE: To determine whether Dexon mesh, closely applied to the kidney, provides purchase for sutures to permit bladder/parenchymal apposition on autotransplantation and that, if this line of apposition were some distance from but surrounding renal papillae, urothelium would proliferate to cover exposed parenchyma to form a widely patent lumen; this should facilitate removal of the whole of an upper tract collecting system, retaining renal parenchyma alone. MATERIALS AND METHODS: To test this possibility and explore the practicability of the concept, nine dogs underwent bilateral nephrectomy followed by unilateral autotransplantation: the other kidney was discarded. Because the canine renal pelvis is intrarenal, the ureter was stretched maximally before passing fine scissors into the renal hilum to transect the collecting system as close to the kidney as possible in six of the nine dogs. In the remaining three dogs, partial nephrectomy was performed with division of the calyceal necks under vision. Thinned bladder wall was sutured to Dexon mesh some distance from the collecting tubules; omentum was applied to the suture line. RESULTS: Three dogs were killed prematurely at < 2 weeks because of perioperative complications. Four were killed at 2, 4, 5 and 8 weeks and two at 12 months. Dexon mesh proved to be an effective anchoring fabric, providing close apposition of bladder wall and parenchyma. There was no adhesion of the kidney to peritoneal contents. Urothelial proliferation to cover exposed parenchyma occurred early and by 12 months, a thin stroma was interposed between parenchyma and epithelium. The kidney was preserved in all but one removed electively, this dog having both cystitis and pyelonephritis at 12 months. CONCLUSIONS: This study showed that autotransplantation of a kidney after removal of its collecting system and advancement of thinned bladder wall to renal parenchyma is practicable, with regenerated urothelium bridging the deficiency by covering exposed parenchyma, to create a widely patent lumen.

Animals↗
PubMed1997

Intussuscepted partial-thickness ileal valve in continent urinary diversion.

OBJECTIVE: This study was undertaken to ascertain the feasibility of fashioning a nipple valve from partial-thickness ileum and to assess the competence and durability of that valve. The approach employed was designed to circumvent the necessity for considerable lengths of bowel to be committed to valve formation and to avoid the tendency for desusception, present with other forms of nipple valves. METHODS: A technique in which a subterminal segment of partial-thickness ileum was 'skinned' circumferentially of serosa and muscularis propria and then intussuscepted to form a continent nipple-valve mechanism was studied for up to 4 months in 10 dogs. The intussuscepted partial-thickness ileal valve was in continuity with a terminal ileal segment sutured flush with skin and, internally, with another segment laid open and anastomosed to the bladder. RESULTS: All valves were competent, withstanding intravesical pressures up to 90 cm H2O. Six dogs were catheterized, without difficulty, twice daily up to 104 days. The valve mucosal surfaces were smooth due to a loss of plicae circulares, and, between 'back-to-back' submucosal layers, a fine stroma developed. CONCLUSIONS: This simple technique, which is frugal in its use of bowel, provided a robust and effective ileal continence mechanism. Furthermore, because of denervation and interposing fibrous tissue, this nipple valve is considered most unlikely to desuscept subsequently. The intussuscepted partial-thickness ileal valve approach is recommended now for clinical evaluation.

Anastomosis, Surgical↗
PubMed1996

Abnormal prostatic cells in ejaculates from men with prostatic cancer--a preliminary report.

OBJECTIVE: To relate findings from a novel approach, ejaculate cytology, to the established reference, histopathology from transrectal ultrasonography (TRUS)-guided prostatic biopsies, in patients at risk of having prostatic cancer on the basis of an abnormal digital rectal examination (DRE) and/or an elevated serum prostate specific antigen (PSA). PATIENTS SUBJECTS AND METHODS: Thirty-seven men suspected of having prostatic carcinoma provided ejaculate specimens which were collected in Hanks solution. The specimens were centrifuged to form a pellet from which smears were made for cytological examination. Immunohistochemical staining for PSA and prostatic acid phosphatase (PAP) were performed on embedded blocks of these cells. TRUS-guided sextant biopsies were performed for histological specimens using standard clinical procedures. A control group of 32 men < 30 years of age, with no family history of prostatic cancer, also produced specimens of ejaculate which were processed similarly. RESULTS: Frankly malignant and atypical prostatic cells were identified in ejaculate specimens from 14 of the 37 patients. Of 12 patients with TRUS biopsies positive for malignancy, nine (75%) had abnormal cells in their ejaculates. Furthermore, five of 25 patients with negative biopsies for adenocarcinoma also had abnormal ejaculate cytology; two of these five patients had high-grade prostatic intra-epithelial neoplasia (PIN). In the control group, no PSA- or PAP-positive prostatic epithelial cells were identified. Normal prostatic cells were not seen in any of the ejaculate specimens examined. CONCLUSIONS: These results indicate that ejaculate cytology, which is a non-invasive and easily repeated investigation, may prove to be a useful approach in the early detection of cancer of the prostate. However, its value in this role, together with the clinical significance of cytological findings, needs to be established, especially in relation to PSA and TRUS biopsy.

Acid Phosphatase↗
PubMed1995

Urological tumours: recent changes.

The principles of urology and all surgical disciplines were, not too long ago, considered to be rigidly and permanently established. In this exciting era of change, the foundations of urology are now set on shifting sands rather than the bedrock of the past. The pertinent issue clinically is making discriminating evaluations of new developments so that, where appropriate, they are able to be incorporated into treatment practices to improve patient care. In this overview, a personal appreciation of the more important practical and conceptual changes relating to prostatic, urothelial and renal carcinoma during the past 5 to 10 years is presented.

Humans↗
PubMed1994

Identification of a potentially radiosensitive subgroup among patients with breast cancer.

BACKGROUND: The description of genes and genetic syndromes, such as ataxia-telangiectasia, that predispose some women to breast cancer will provide greater insight into the genetic basis of cancer susceptibility. PURPOSE: Our goal was to establish cell lines from patients with breast and bladder cancers, to screen for enhanced levels of radiation-induced arrest in the G2 phase of the cell cycle such as is observed in ataxia-telangiectasia heterozygotes, and to correlate G2 arrest with other prognostic indicators of these cancers and in vivo radiosensitivity. METHODS: Epstein-Barr virus-transformed lymphoblastoid cells were established from 108 female patients with breast cancer and 24 age-matched female control subjects, and from 45 patients with bladder cancer and 18 age-matched control subjects. Cells were exposed to 3 Gy of gamma radiation, and the percentages of cells in G1 and G2 phases were determined at 18 and 24 hours after irradiation by fluorescence-activated cell sorter analysis. Postirradiation delay in G2 phase was determined by calculating the percentage of cells in G2 and by using the ratio G2/G1. RESULTS: When we determined the percentage of cells in G2 phase at 18 hours after irradiation in 108 lymphoblastoid cells from breast cancer patients, we observed an increase of between 3% and 38% in the number of cells in G2 phase in comparison with cells that were not irradiated. Comparison with previous G2-phase arrest data for ataxia-telangiectasia heterozygotes using a cutoff point at 29% delay demonstrated that 20% and 8% of the breast cancer cell lines of the case patients and control subjects, respectively, fell into that category (P < .001). At the same time after irradiation, it was not possible to distinguish between bladder cancer cell lines (7%) and those of the corresponding control group (6%). Assessment of radiation effects by G2/G1 ratio showed that 18% of the breast cancer patients and 8% of the control subjects were in the high range. When G2 arrest was correlated with other prognostic factors, we found that case patients with a greater G2 block were more likely to have had a family history of breast cancer (P < .006) and more aggressive tumors when assessed by number of involved lymph nodes (P < .002) and tumor size (P < .05). Furthermore, an adverse response to radiotherapy was observed in a group of patients with high G2 arrest. IMPLICATIONS: While the postirradiation increase in G2-phase arrest in cells from breast cancer patients observed in this study may indicate genetic heterozygosity for ataxia-telangiectasia, it might also reflect other genetic abnormalities important to breast cancer.

Adult↗
PubMed1994

Accumulation of HL-60 leukemia cells in G2/M and inhibition of cytokinesis caused by two marine compounds, bistratene A and cycloxazoline.

The effects on the cell cycle of two biologically active compounds, bistratene A and cycloxazoline, from the marine ascidian Lissoclinum bistratum were studied in HL-60 human leukemia cells using flow cytometry. Both compounds were shown to cause an apparent accumulation of cells in the G2/M phase. This effect was shown to be both time- and dose-dependent. At the longer time points (30 and 48 h after addition of the compounds) polyploidy was apparent. The fate of cells labeled in the S phase with 5-bromo-2'-deoxyuridine (BrdUrd) was analysed using a bivariate BrdUrd/PI (propidium iodide) technique. Bistratene A and cycloxazoline treatment prevented the majority of BrdUrd-labeled cells from progressing through to the G1 phase. Approximately 50% of the cells were delayed at G2/M, and a significant proportion of cells appeared to be polyploid. Light and electron microscopy revealed the presence of multinucleated cells accounting for the apparent polyploidy. The progression of cells out of the G1 phase was also examined by synchronising cells with mimosine and releasing them from mimosine block in the presence of bistratene A. There was no evidence of a block at the G1/S phase transition or through the S phase since DNA synthesis was not inhibited. The mechanism by which these compounds interfere with cytokinesis is presently unknown but, in the case of bistratene A, may be linked to altered phosphorylation of cellular proteins involved in cell-cycle control.

Acetamides↗
PubMed1994

Mucin expression by transitional cell carcinomas of the bladder.

OBJECTIVE: To examine the presence of a membrane-associated and secreted mucin (MUC1) and a secreted gel-forming mucin (MUC2) in normal and malignant urothelium. MATERIALS AND METHODS: Sections were obtained from archival paraffin blocks from 11 patients with nonmalignant urological conditions and 89 patients with transitional cell carcinomas (TCC). Mucin expression was examined by immunohistochemistry using monoclonal antibodies BC2 and 4F1, reactive with epitopes on the protein core of MUC1 and MUC2 respectively. RESULTS: In normal urothelium MUC1 was limited predominantly to the apical membranes of the umbrella cell layer. MUC1 was present in all cases of TCC, and the pattern of expression divided into three categories: luminal membrane staining only, luminal plus cytoplasmic staining of intermediate +/- basal layers, or staining of only isolated cells or cell groups. These staining patterns were significantly associated with both tumour grade and stage (P < 0.001), with cytoplasmic staining more prevalent in higher grade and stage tumours. MUC2 was not detected in normal urothelium, and was present in 40% of cases of TCC, characterized by intense granular cytoplasmic staining. No association between MUC2 expression and either tumour grade or stage was demonstrated. CONCLUSION: MUC1 mucin was expressed by both normal and malignant urothelium, with increased expression characteristic of higher grade and stage tumours. MUC2 expression was found in 40% of tumours but not in normal urothelium. The role of these mucins in the biology of the bladder requires further investigation.

Aged↗
PubMed1994

Immunohistological expression of p53 in primary pT1 transitional cell bladder cancer in relation to tumour progression.

OBJECTIVE: To determine whether p53 expression is a marker of tumour progression in superficially invasive (pT1) transitional cell carcinoma of the bladder. PATIENTS AND METHODS: The immunohistochemical status of the p53 protein in 28 pT1 primary bladder cancers was determined on frozen tissue and archival paraffin block sections using three primary antibodies (CM-1, PAB1801 and D07). The findings were compared with the patients' progress. All the patients, except for those who died during the course of the study, were followed up by check cystoscopy for a minimum of 2 years. RESULTS: Immediately adjacent frozen sections stained identically in 10 of 16 cases for CM-1 and PAB1801. For paraffin sections, identical staining patterns were seen for PAB1801 and D07 in 21 of 26 sections. However, inter- and intra-tumour staining for p53 was very variable, even with the same antibody. The heterogeneity of p53 positive cell distribution in the tumours indicates potential for significant sampling errors if random sections are chosen as representative of p53 status. CONCLUSION: The p53 status of the primary tumours did not relate to patient outcome. The results obtained do not support the use of immunohistological p53 expression as a discriminating prognostic indicator in pT1 transitional cell carcinoma of the bladder.

Biomarkers, Tumor↗
PubMed1994

The invaginated sleeve technique for a continent cystostomy--five years' clinical experience.

OBJECTIVE: To evaluate the invaginated sleeve technique for continent cystostomy in humans. PATIENTS AND METHODS: Over the past 5 years six patients have undergone this operative procedure. An essential principle utilized in the technique is the property of partial thickness bladder grafts to stretch longitudinally, so that each tubularized pedicle graft could be directed transmurally and extended extravesically to reach the skin of the lower abdominal wall. As a consequence a urothelial-lined tract, both extravesically and through the bladder wall, was provided for intermittent self-catheterization. RESULTS: Apart from one woman, whose cystostomy tract was disrupted by inappropriate catheterization in the immediate post-operative period to attempt to stop leakage through exposed fenestrations in the suprapubic stent, this procedure provided robust, continent catheterizing routes for all patients for periods of 63, 52, 12, 7 and 1.5 months respectively. Two patients developed discrete stenoses at their mucocutaneous junctions at 3 and 5 months which were corrected easily. CONCLUSION: This simple, minimally morbid technique, which avoids the use of non-urinary tract epithelial structures and maintains bladder capacity, is strongly recommended for patients who need to practise clean intermittent self-catheterization and for whom the urethral route is impracticable.

Catheters, Indwelling↗
PubMed1993

The adverse effect of fibrin-clot inhibiting drugs on intravesical bacillus Calmette-Guérin efficacy for superficial bladder cancer.

Bacillus Calmette-Guérin (BCG) is currently thought to act as a biological immune modifier in effecting antitumour activity. Recent evidence suggests that BCG binding to fibronectin (FN), a tissue glycoprotein, may be a prerequisite step in initiating this response. Drugs inhibiting the availability of exposed FN in the bladder after urothelial disruption may adversely affect the efficacy of BCG. Data are presented of 45 patients with tumour limited to mucosa (pTa) or carcinoma in situ (CIS) given intravesical BCG therapy, with (group 1) or without (group 2) fibrin clot-inhibiting drugs concurrently during treatment. The success rate of 11.1% for group 1 (1/9) patients was significantly less than that of 69.4% for group 2 (25/36), (chi 2 = 7.79, P < 0.01 Fisher's exact test) supporting the suggestion that the concurrent administration of fibrin-clot inhibiting drugs may adversely affect the outcome of BCG therapy.

Administration, Intravesical↗
PubMed1992

An immunohistological demonstration of c-erbB-2 oncoprotein expression in primary urothelial bladder cancer.

Sections of formalin-fixed, paraffin-blocked tissue from 116 primary transitional cell carcinomas were stained immunohistochemically using a polyclonal antibody against the c-erbB-2 oncoprotein. Positive staining of cell membranes, known to correlate with gene amplification, was seen in 22 (19%) of the 116, with variable staining from tumour to tumour and within tumours themselves. Consistent with its mooted value as a prognosticator in bladder cancer, the c-erbB-2 oncoprotein was detected in 13 (of 40) grade III and 9 of the 26 muscle-invasive tumours examined compared to 1 (of 25) grade I and 6 (of 66) mucosa only (pTa) lesions. These results support further examination of c-erbB-2 expression in bladder cancer.

Aged↗
PubMed1990

Evaluation of a bladder advancement extension graft technique.

A bladder advancement extension technique was developed to provide a reliable method for creating an extra length of bladder in continuity, so that Boari flap and psoas hitch procedures, with or without the extension grafts, would reach sufficiently high routinely to permit these procedures to become the preferred practical options for accommodating significant proximal ureteric deficiencies or for replacing the whole of the ureter. In addition, an omentally supported bladder graft was established separately for use in a "retrieval" procedure should extension graft stenosis develop subsequently. Of 10 dogs studied for 6 months, 2 required "retrieval" pedicled island patch grafts for stenoses at extension graft/upper ureteric junctions. Healthy urothelium with muscle in the walls was seen in grafts resting on their supporting omental beds. At the time of sacrificing, none of the operated upper tracts was obstructed.

Animals↗
PubMed1990

Transferrin receptor expression in primary superficial human bladder tumours identifies patients who develop recurrences.

A group of 65 patients with superficial bladder carcinoma was followed for 2 years and tumour recurrence rate was correlated both with transferrin receptor status of the initial primary tumour and with the results of voided urine cytology. Nine of 24 patients with transferrin receptor negative tumours had recurrences compared with 30 of 41 patients with transferrin receptor positive tumours. This difference was highly significant. Urine cytology at presentation was also predictive of further tumour formation: of 30 patients who were transferrin receptor positive and had positive urine cytology, 25 developed recurrences.

Adult↗
PubMed1990

Application of the invaginated sleeve technique to form a continent ileal urostomy.

The invaginated sleeve technique has been adapted to produce a continent urostomy, able to be catheterised, from a length of canine ileum measuring less than 10 cm. Of the 4 dogs catheterised twice daily up to 13 weeks, narrowing was present in the extra-ileal segment in 2, but all intra-ileal double-barrelled lumina remained healthy with preservation of bowel mucosa.

Animals↗