A.J. Cooper

The effects of dietary lycopene supplementation on human seminal plasma

To investigate whether lycopene levels in blood and seminal plasma increase after dietary supplementation with a natural source of the compound, and whether any potential increase of lycopene levels in semen translates into increased free‐radical trapping capacity in the seminal plasma.

Epirubicin and meglumine ?-linolenic acid: a logical choice of combination therapy for patients with superficial bladder carcinoma

BACKGROUND Anthracyclines have been established as first-line drugs for intravesical use in the treatment of patients with superficial bladder carcinoma, although they result only in a modest reduction in tumor recurrence rates. The essential fatty acid γ-linolenic acid (GLA) also is an effective cytotoxic agent against superficial bladder carcinoma when it is applied topically. The objective of this study was to assess the efficacy of combined epirubicin and GLA with the purpose of developing a suitable model for modification of existing intravesical regimens. METHODS The human urothelial carcinoma cell lines MGH-U1 and RT112 were used in standard cytotoxicity assays and were exposed to meglumine GLA (MeGLA) and epirubicin in two-dimensional concentration matrices. A thiozolyl blue (methyl-thiazoldiphenyl tetrazolium) assay was used to determine residual cell biomass. Drug interaction was quantified by median-effect analysis software (CalcuSyn®), and the evaluation of drug uptake utilized fluorescence confocal microscopy (FCM) and flow cytometry. RESULTS MeGLA caused a significant enhancement of anthracycline uptake, viewed by FCM, from 92 fluorescence units to 222 fluorescence units (P < 0.001). Flow cytometry confirmed the increased drug uptake and showed that the mean epirubicin content per cell increased from 23 to 57 units and from 8 to 24 units for MGH-U1 and RT112 cells, respectively (99% confidence interval < 0.3). This resulted in improved cytotoxicity, and it was shown that the drugs acted synergistically with all but the highest MeGLA concentrations. CONCLUSIONS The efficacy of epirubicin was enhanced significantly when it was used in combination with most concentrations of MeGLA (< 300 μg/mL), and the two agents acted synergistically. There was a corresponding increase in epirubicin uptake by cells under these conditions. At high MeGLA concentrations, however, anthracycline solubility was compromised, and drug synergy was lost. Cancer 2003;97:71–8.

The craniocervical junction following successful haematopoietic stem cell transplantation for mucopolysaccharidosis type I H (Hurler syndrome)

Mucopolysaccharidosis I Hurler (MPS IH) is a progressive multisystemic disorder caused by alpha-L-iduronidase deficiency. First choice of treatment in MPS IH children is haematopoietic stem cell transplantation (HSCT). The effect of HSCT has been shown to have limited influence on skeletal manifestations by poor penetration of musculoskeletal tissues by the enzyme derived from donor leucocytes. Aim of this study was to investigate the effect of HSCT on the craniocervical junction (CCJ) in Hurler patients. We analysed retrospectively sequential magnetic resonance imaging (MRI) scans of 30 patients with Hurler disease treated by HSCT since 1982 at the Royal Manchester Children’s Hospital, UK, in order to determine whether the patients suffer from dens hypoplasia. Results were compared with biochemical and clinical characteristics: Enzyme activity (EA), chimerism, urinary glycosaminoglycan (GAG) excretion and neurological status. Investigations were part of standard clinical procedures. Results are descriptive in presentation. In 26/30 patients a determination of odontoid hypoplasia was feasible. The majority showed a normal dens length and an increase with age. Only 3/26 revealed a dens hypoplasia. One of them had only partial donor engraftment (DE) with reduced EA, one of them suffered from chronic graft versus host disease (GVHD). One patient with only partial DE and reduced EA presented with initial dens hypoplasia until preadolescence but normalized later on. There may be a trend towards lower EA and the occurrence of DH in transplanted MPS patients - perhaps the dosage of enzyme plays a role in the correction of skeletal complications in this patient group. HSCT patients with incomplete DE and therefore lower EAs may require special attention and care.

In vitro inhibition of angiogenesis by prostasomes

Prostasomes are biologically active organelles that are secreted by human prostate epithelial cells, and it is believed that they have a role in prostatic disease. We studied the effect of prostasomes on the human umbilical vein endothelial cell (HUVEC)/Matrigel model of angiogenesis, and the association of labelled prostasomes with HUVECs. The growth inhibitory effect of prostasomes on HUVECs was assayed by spectrophotometric measurement of residual biomass. Preparations of HUVECs on a Matrigel base were exposed to prostasomes, and the development of capillary-like networks was quantified. Prostasomes were labelled with PKH-26, and cultured with HUVECs. Prostasomes were not shown to have a significant effect on HUVEC survival. Angiogenesis assays showed inhibition. The PKH-26-labelled particles were shown to have adhered to the HUVECs. This study adds the inhibition of an in vitro correlate of angiogenesis to the known actions of prostasomes.

Cellular resistance to mitomycin C is associated with overexpression of MDR‐1 in a urothelial cancer cell line (MGH‐U1)

Objective To compare multidrug resistance (MDR)‐1 and MDR‐3 gene expression in a new urothelial cancer cell line (MGHU‐1, with resistance to mitomycin C) against controls and the established (epirubicin‐resistant) MDR clone, and to correlate MDR with cytotoxicity data.

Intravesical meglumine gamma-linolenic acid in superficial bladder cancer: An efficacy study

OBJECTIVES Gamma-linolenic acid (GLA) is known to be cytotoxic to malignant cells. We assessed the efficacy of the novel intravesical formulation, meglumine gamma-linolenic acid (MeGLA), in a phase II trial, in patients with recurrent, superficial bladder cancer. PATIENTS AND METHODS Thirty patients with recurrent, superficial transitional cell carcinoma (TCC) were recruited. The tumour pattern was recorded at flexible cystoscopy. Patients received a single intravesical instillation of 50ml of either 50mg (1mg/ml) (15 patients), or 125mg (2.5mg/ml) (15 patients) of MeGLA in water, retained for one hour. At subsequent cystoscopy, the tumour patterns were recorded, prior to undertaking routine cystodiathermy. Biopsies were obtained for histological assessment. Responses were divided into complete, partial or none. RESULTS All 30 patients retained the drug for 1 hour without significant local or systemic side effects. There were 4 (13%) complete responses, 9 (30%) partial responses, and 17 (57%) non-responders. Histology showed no evidence of damage to surrounding urothelium. CONCLUSIONS Our data confirms the safety and tolerability of MeGLA, which is consistent with findings from a previous phase I trial. A response rate of 43% also indicates that MeGLA has a significant cytotoxic effect against TCC and the results are similar to those obtained using standard, single-dose, intravesical regimens.

Molecular analysis of the β-globin gene cluster haplotypes in a Sudanese population with sickle cell anaemia

Introduction:  Sudan has a multiethnic population with a high frequency of Hb S, but little is known about the βS haplotypes in this population.

Prostate cells exposed to lycopene in vitro liberate lycopene‐enriched exosomes

To investigate whether cellular exosomes liberated by prostatic cell lines in culture might be acting as the transport vehicles for the dietary antioxidant lycopene, known to be sequestered in the prostate gland and to reduce the risk of developing benign prostatic hyperplasia (BPH) and prostate cancer; its subsequent secretion into seminal plasma also confers protection to spermatozoa against oxidative free‐radical damage.

Prostasomes: a role in prostatic disease?

Prostasomes are membrane-bound secretory vesicles, in the nanometre diameter range, secreted by the prostatic ductal epithelium [1] into the lumen, where they form part of the ejaculate. Although known to have specific biological properties, their physiological role and overall significance remain far more debatable. There has been comparatively little written about these structures since their detection two decades ago, and this may help explain the relative lack of awareness of prostasomes within the urological community.

Lycopene treatment of prostate cancer cell lines inhibits adhesion and migration properties of the cells.

Background: Consumption of lycopene through tomato products has been suggested to reduce the risk of prostate cancer. Cellular adhesion and migration are important features of cancer progression and therefore a potential target for cancer interception. In the present study we have examined the in vitro effect of lycopene on these processes. Methods: Prostate cancer cell lines PC3, DU145 and immortalised normal prostate cell line PNT-2 were used. The adhesion assay consisted of seeding pre-treated cells onto Matrigel™, gently removing non-adherent cells and quantitating the adherent fraction using WST-1. Migratory potential was assessed using ibidi™ migration chamber inserts, in which a cell-free zone between two confluent areas was allowed to populate over time and the migration measured. Results: 24 hour incubation of prostate cell lines with 1.15µmol/l lycopene showed a 40% reduction of cellular motility in case of PC3 cells, 58% in DU145 cells and no effect was observed for PNT2 cells. A dose related inhibition of cell adhesion to a basement membrane in the form of Matrigel™ was observed in all three cell lines and it reached statistical significance for PC3 and PNT2 cells at lycopene concentrations ≥1.15µmol/l. However, in case of DU145, only a concentration of 2.3µmol/l showed a significant reduction. Conclusion: This in vitro investigation indicates that lycopene can influence the cell adhesion and migration properties of cancer cells at a dose which is arguably achievable in patients. The results of our study expand our understanding of a chemo preventive role of lycopene in prostate cancer.