Tumour cell lysate-loaded dendritic cell vaccine induces biochemical and memory immune response in castration-resistant prostate cancer patients.

BACKGROUND: Recently, we produced a tumour antigen-presenting cells (TAPCells) vaccine using a melanoma cell lysate, called TRIMEL, as an antigen source and an activation factor. Tumour antigen-presenting cells induced immunological responses and increased melanoma patient survival. Herein, we investigated the effect of TAPCells loaded with prostate cancer cell lysates (PCCL) as an antigen source, and TRIMEL as a dendritic cell (DC) activation factor; which were co-injected with the Concholepas concholepas haemocyanin (CCH) as an adjuvant on castration-resistant prostate cancer (CRPC) patients. METHODS: The lysate mix capacity, for inducing T-cell activation, was analysed by flow cytometry and Elispot. Delayed-type hypersensitivity (DTH) reaction against PCCL, frequency of CD8(+) memory T cells (Tm) in blood and prostate-specific antigen (PSA) levels in serum were measured in treated patients. RESULTS: The lysate mix induced functional mature DCs that were capable of activating PCCL-specific T cells. No relevant adverse reactions were observed. Six out of 14 patients showed a significant decrease in levels of PSA. DTH(+) patients showed a prolonged PSA doubling-time after treatment. Expansion of functional central and effector CD8(+) Tm were detected. CONCLUSION: Treatment of CRPC patients with lysate-loaded TAPCells and CCH as an adjuvant is safe: generating biochemical and memory immune responses. However, the limited number of cases requires confirmation in a phase II clinical trial.

Cell grazing and Haemonchus contortus control in sheep: lessons from a two-year study in temperate Western Europe.

Managing infections of sheep with anthelmintic resistant gastrointestinal nematodes (GIN) is a major challenge for sheep producers in Western Europe. New methods of grazing management have been poorly explored as a component of an integrated and sustainable control of these parasites. Therefore, the purpose of this study was to evaluate the effect of two different types of grazing systems of sheep (intensive cell grazing versus conventional rotational grazing) on GIN infections over two years in a farm located in a temperate environment of Western France. When considering the whole study, the type of grazing system did not influence significantly the intensity of egg excretions of adult ewes even if the proportion of ewes excreting high numbers of GIN eggs was higher in cell grazing system than in rotational grazing system. The most striking result of this survey was the effect of grazing system on the GIN species composition harbored by ewes and by their lambs: with time, the proportions of H. contortus infections were lower in cell grazing system than in rotational grazing system. In conclusion, the cell grazing system, as implemented in this study, could limit the importance of this highly pathogenic nematode species in sheep.

Proapoptotic effect of endocannabinoids in prostate cancer cells.

In the early stages, prostate cancer is androgen­ dependent; therefore, medical castration has shown significant results during the initial stages of this pathology. Despite this early effect, advanced prostate cancer is resilient to such treatment. Recent evidence shows that derivatives of Cannabis sativa and its analogs may exert a protective effect against different types of oncologic pathologies. The purpose of the present study was to detect the presence of cannabinoid receptors (CB1 and CB2) on cancer cells with a prostatic origin and to evaluate the effect of the in vitro use of synthetic analogs. In order to do this, we used a commercial cell line and primary cultures derived from prostate cancer and benign prostatic hyperplasia. The presence of the CB1 and CB2 receptors was determined by immunohistochemistry where we showed a higher expression of these receptors in later stages of the disease (samples with a high Gleason score). Later, treatments were conducted using anandamide, 2-arachidonoyl glycerol and a synthetic analog of anandamide, methanandamide. Using the MTT assay, we proved that the treatments produced a cell growth inhibitory effect on all the different prostate cancer cultures. This effect was demonstrated to be dose-dependent. The use of a specific CB1 receptor blocker (SR141716) confirmed that this effect was produced primarily from the activation of the CB1 receptor. In order to understand the MTT assay results, we determined cell cycle distribution by flow cytometry, which showed no variation at the different cell cycle stages in all the cultures after treatment. Treatment with endocannabinoids resulted in an increase in the percentage of apoptotic cells as determined by Annexin V assays and caused an increase in the levels of activated caspase-3 and a reduction in the levels of Bcl-2 confirming that the reduction in cell viability noted in the MTT assay was caused by the activation of the apoptotic pathway. Finally, we observed that endocannabinoid treatment activated the Erk pathway and at the same time, produced a decrease in the activation levels of the Akt pathway. Based on these results, we suggest that endocannabinoids may be a beneficial option for the treatment of prostate cancer that has become nonresponsive to common therapies.

Use of permanent interstitial temperature self-regulating rods for ablation of prostate cancer.

BACKGROUND AND PURPOSE: We have examined a new technique for the thermal treatment of prostate cancer involving the percutaneous placement of small magnetic alloy rods into the gland in a procedure analogous to the placement of permanent brachytherapy seeds. When the patient and the implanted rods are placed in an extracorporeal alternating magnetic field, the rods heat and can produce tissue necrosis. The rods are temperature self-regulating and produce temperatures no higher than their preprogrammed regulation value. The rods are biocompatable permanent implants, comprised of cobalt and palladium. MATERIALS AND METHODS: We examined rods that heat to 55 degrees, 60 degrees, and 70 degrees C. Patients are treated for 1-hour sessions and easily have multiple sessions. RESULTS: Isolated rods produced little or asymmetrical necrosis. However, arrays of 70 degrees C rods placed within 1 cm of each other caused consistent necrosis between the rods. The temperature at the edge of the array dropped off quickly, and therefore, the rods can be placed within 2 mm of the capsule even posteriorly near the rectum. CONCLUSIONS: Arrays of high-temperature rods can be employed for tissue ablation, while lower-temperature arrays could be used to produce hyperthermia in order to achieve synergism with adjuvant radiation therapy. The technique is applicable in an outpatient setting.

Neoadjuvant and salvage chemotherapy with cisplatin (CDDP) and 5-fluorouracil (5-FU) in cervical carcinoma.

A regime of cis-platin (CDDP) (80 mg/m2) and 5-fluorouracil (5-FU) (1000 mg/m2/day, day 1 to 5) repeated after 21 days, was prospectively analyzed in 12 advanced epidermoid cervical cancers as first treatment, prior to radiotherapy (neoadjuvant chemotherapy), and in 30 cases of progressive, recurrent or metastatic disease after radiotherapy (salvage chemotherapy), in order to evaluate efficacy and toxicity. Among the 10 evaluable neoadjuvant cases we observed 2 complete, 7 partial responders and 1 stabilized. They all achieved a complete response after radiotherapy and 6 remain alive after 18 to 72 months. None of the salvaged patients achieved a complete response and only 26.9% responded partially. Only one case, though fatal, of myelodepression was found in the neoadjuvant group. Conversely, 70% of salvages showed some grade of myelodepression, being severe or extremely severe in 23.3%, with another case of death. Neoadjuvant chemotherapy with CDDP and 5-FU seems promising in advanced cervical carcinoma and is acceptably well tolerated. In contrast, salvage therapy with the same regime yields worse results and is much more toxic.

Aberrant epigenetic regulation of bromodomain BRD4 in human colon cancer.

The bromodomain protein BRD4 is involved in cell proliferation and cell cycle progression, primarily through its role in acetylated chromatin-dependent regulation of transcription at targeted loci. Here, we show that BRD4 is frequently downregulated by aberrant promoter hypermethylation in human colon cancer cell lines and primary tumors. Ectopic re-expression of BRD4 in these colon cancer cell lines markedly reduced in vivo tumor growth, suggesting a role of BRD4 in human colon cancer.

AGNES: A technique for determining the concentration of free metal ions. The case of Zn(II) in coastal Mediterranean seawater.

Absence of Gradients and Nernstian Equilibrium Stripping (AGNES) is a recently suggested electroanalytical technique designed for the determination of the free concentration of heavy metals (such as Zn, Cd or Pb) which is here developed and applied to seawater samples. A key improvement for the implementation of AGNES with complex matrices is the development of a new blank, called the shifted blank (presented in this work for the first time), which can be applied to the same sample where the measurement is intended. The careful selection of the required parameters for the determination of the free Zn concentration (or activity) at the nanomolar level is described in detail. The methodology has been validated with a synthetic solution containing Zn and nitrilotriacetic acid (NTA) and then applied, as a first case, to two coastal seawater samples taken close to Barcelona and Tarragona (Catalonia, North-Eastern Spain) finding values in the range of 1-3nM, representing around 25% of total Zn. This technique can, in the near future, be crucial in helping to elucidate the role of the free zinc(II) concentration in natural waters.

Androgen regulation of glycosidase secretion in epithelial cell cultures from human epididymis.

The human epididymis and its secretions actively promote sperm fertilizing capacity and provide protection for spermatozoa against harmful influences. Among epididymal secretions, glycosidases have been recently studied and associated with molecular changes on the sperm surface. In the present work, we studied the influence of different concentrations of testosterone, dihydrotestosterone and cyproterone acetate on the secretion of alpha-glucosidase, N-acetyl-glucosaminidase, beta-glucuronidase and alpha-mannosidase by isolated and cultured epithelial cells from human caput, corpus and cauda epididymides. Cell cultures were obtained from aggregates of isolated tubule fragments plated on extracellular matrix-covered multi-well plates. Activities of the glycosidases were measured in conditioned culture media and were higher in the distal regions of the epididymis. Testosterone and dihydrotestosterone significantly increase the enzyme secretion in a concentration-dependent manner. This increase was higher in corpus and/or cauda than in caput epididymis. Cyproterone acetate caused a dose-dependent decrease in glycosidase secretion in cultures from all epididymal regions. It is concluded that the secretion of epididymal glycosidases is regulated by androgen, being stimulated by dihydrotestosterone and testosterone and inhibited by the androgen antagonist cyproterone acetate.

Glutathione-related enzymes in cell cultures from different regions of human epididymis.

Protection of maturing sperm from potential endogenous or exogenous harmful substances during their transit throughout the epididymis is a critical event. The authors studied the activity of gamma-glutamyl transpeptidase (GGT) and glutathione S-transferase (GST), and glutathione (GSH) levels in epithelial cell cultures from human caput, corpus, and cauda epididymides. Tissue was obtained from patients undergoing therapeutic orchidectomy for prostatic cancer. Enzymatic activity was measured in conditioned media and cellular fractions. Androgen influence was also evaluated. Both enzymatic activities were found in cellular homogenates and conditioned media from cultures of all epididymal regions. GGT activity was highest in cultures from cauda epididymis, both in conditioned media and cell fractions, while GST activity did not show regional differences in conditioned media, but exhibited higher activity in cell homogenates from cauda cultures than those obtained from corpus and caput epididymis. GSH level showed no regional difference in cell homogenates and it could not be detected in conditioned media by the method used. Presence of different concentrations of dihydrotestosterone (DHT) had no influence neither on the enzymatic activities nor GSH concentration. The results indicate that GGT and GST are present along the human epididymis and a fraction or isoform of these enzymes might be secreted to the luminal fluid to play a detoxificative role in sperm maturation.

The role of the epididymis in human infertility.

This review describes the role of the epididymis in human infertility, by analysing the results of epididymovasostomies which confirm that the more distal the site of obstruction the greater the chance of fertility. The use of epididymal spermatozoa for in vitro fertilization (IVF) yielded poor results in contrast to intracytoplasmic sperm injection using either epididymal or testicular spermatozoa. The nature of the pathology causing obstructive azoospermia is examined reviewing in particular the possible role of mercury toxicity in Young's syndrome. This review describes the results of studies that show that the level of obstruction within the epididymis is correlated with the presence of sperm antibodies and distal obstructions are associated with the presence of sperm antibodies. The demonstration that necrozoospermia decreases with increased sperm transport through the epididymis, when combined with the observation of normal testicular sperm morphology, represents the basis for the disorder termed epididymal necrozoospermia. However, to date the nature of the epididymal pathology causing this disorder remains obscure.

Polimorfismo de citocromo P-450(CYP1A1*2A) como factor de riesgo y pronóstico del cáncer de próstata / Polymorphism of cytochrome P450 (CYP1A1 * 2A) as a risk factor andprognosis of prostate cancer

INTRODUCCIÓN: Polimorfismos en enzimas de biotransformación hansido asociados como factores de riesgo de cáncer prostático (CaP), aunque su papel como marcadores de pronóstico no está totalmente validado. El objetivo de este trabajofue estudiar en un grupo de pacientes sometidos a tamizaje, la utilidad del polimorfismo de CYP1A1 en el diagnóstico y sobrevida de pacientes chilenos con CaP.MATERIALES Y MÉTODOS: 255 controles y 234 casos con CaP fueron incluidos en programa de tamizaje en CONAC entre1995 y 2004. De muestras de sangre periférica se obtuvo DNA genómico y determinó polimorfi smo de CYP1A1*2A. Paraanálisis de susceptibilidad se usaron modelos de regresión logística uni y multivariado. Los pacientes con CaP fueron seguidos por 8,76 años en promedio, determinando sobrevida global y cáncer – específica a través de curvas de Kapplan-Meier y test de Log-rank. Se obtuvieron riesgos (Hazard Ratio) ajustados con modelo proporcional de Cox, con IC del 95%...

INTRODUCTION: Polymorphisms in biotransformation enzymes have been associated as risk factors for prostate cancer(CaP), although their role as prognostic markers is not fully validated. The aim of this work was to study in a group ofpatients undergoing screening, utility CYP1A1 polymorphism in the diagnosis and survival of Chilean patients with PCa.MATERIALS AND METHODS: 255 cases and 234 controls with CaP that were included in CONAC screening program between1995 and 2004. Genomic DNA was obtained from samples of peripheral blood and polymorphism of CYP1A1 * 2ª determined.For sensitivity analysis uni and multivariate logistic regression models were used. PCa patients were followed for8.76 years on average, determining overall and cancer–specifi c survival through Kapplan- Meier curves and log-rank test.Risks (Hazard Ratio) adjusted Cox proportional model, with 95% was obtained...

Intrauterine insemination: critical analysis of a therapeutic procedure.

Donor intrauterine insemination with washed spermatozoa (fresh semen) was performed in 36 women (63 cycles) whose husbands had azoospermia due to primary or secondary testicular failure. Simultaneously a control group of 76 couples (156 cycles) with proven fertility, who had recently discontinued mechanical non-hormonal contraception, were encouraged to have sexual intercourse during the fertile period. The age of the women was similar in both groups and the timing criteria were also similar. The pregnancy rate per woman was 50% in the donor insemination group and 47.4% in the control group (difference not significant). The pregnancy rate per cycle was 28.6% in the artificial insemination group and 23.1% in the control, natural insemination group (difference not significant). The number of artificial or natural insemination cycles required to achieve pregnancy was similar in the study and control groups. This study suggests that when the inseminating spermatozoa and female partner are normal, as occurs in the donor insemination group, intrauterine insemination is as efficient as natural insemination in achieving pregnancy but is not more successful.

Fertilization rate in couples with unexplained infertility.

A group of 24 couples with unexplained infertility was scheduled for in-vitro fertilization and tubal embryo transfer between May 1989 and September 1990. In the same period, in-vitro fertilization and intrauterine transfer of embryos was planned in a control group of 44 women with tubal infertility. The mean age and duration of infertility were similar in both groups and the same scheme of ovarian stimulation was used. No statistically significant difference was obtained comparing oestradiol levels and numbers of mature oocytes retrieved between the group of patients with unexplained infertility and those with tubal infertility. The fertilization rate of the oocytes obtained from women with unexplained infertility (60.4%) was significantly lower (P less than 0.001) than that of the oocytes obtained from patients with tubal infertility (87.3%). There was no statistically significant difference in the cleavage rates between patients with unexplained infertility and those with tubal infertility. It is concluded that lack of fertilization is an unexplored cause of infertility in couples with unexplained infertility.

Tratamiento de la prostatitis crónica con Kelfiprin / Chronic prostatitis treatment with Kelfiprin.

Se presenta una serie de 18 prostatitis bacterianas y 21 prostatitis no bacterianas tratadas durante 30 dias con Kelfiprin (R). Se reportan resultados exelentes o buenos en el 94% de las prostatitis bacterianas y en el 29% de las no bacterianas