sepharose and Prostatic-Neoplasms

The STEAP1 is a cell-surface antigen over-expressed in prostate cancer, which contributes to tumor progression and aggressiveness. However, the molecular mechanisms underlying STEAP1 and its structural determinants remain elusive.. The STEAP1 was not fully captured with 1.375 M (Butyl), in contrast with interfering heterologous proteins, which were strongly retained and mostly eluted with water. This single step demonstrated higher selectivity of Butyl-Sepharose for host impurities removal from injected crude samples. Co-IP allowed recovering a purified fraction of STEAP1 and contributed to unveil potential physiologically interacting counterparts with the target. A. A method for successfully isolating human STEAP1 from LNCaP cells was provided, avoiding the use of detergents to achieve stability, even outside a membrane-mimicking environment.

Topics: Antigens, Neoplasm; Circular Dichroism; Humans; Immunoprecipitation; Male; Oxidoreductases; Prostatic Neoplasms; Protein Stability; Sepharose

Focal therapy has been proposed as an alternative method to whole-gland treatment for prostate cancer when aiming to reduce treatment side effects. The authors recently validated a radiobiological model which takes into account tumor location and tumor characteristics including tumor cell density, Gleason score, and hypoxia in order to plan optimal dose distributions for focal therapy. The authors propose that this model can be informed using multiparametric MRI (mpMRI) and in this study present a registration framework developed to map prostate mpMRI and histology data, where histology will provide the "ground truth" data regarding tumor location and biology. The authors aim to apply this framework to a growing database to develop a prostate biological atlas which will enable MRI based planning for prostate focal therapy treatment.. Six patients scheduled for routine radical prostatectomy were used in this proof-of-concept study. Each patient underwent mpMRI scanning prior to surgery, after which the excised prostate specimen was formalin fixed and mounted in agarose gel in a custom designed sectioning box. T2-weighted MRI of the specimen in the sectioning box was acquired, after which 5 mm sections of the prostate were cut and histology sections were microtomed. A number of image processing and registration steps were used to register histology images with ex vivo MRI and deformable image registration (DIR) was applied to 3D T2w images to align the in vivo and ex vivo MRI data. Dice coefficient metrics and corresponding feature points from two independent annotators were selected in order to assess the DIR accuracy.. Images from all six patients were registered, providing histology and in vivo MRI in the ex vivo MRI frame of reference for each patient. Results demonstrated that their DIR methodology to register in vivo and ex vivo 3D T2w MRI improved accuracy in comparison with an initial manual alignment for prostates containing features which were readily visible on MRI. The average estimated uncertainty between in vivo MRI and histology was 3.3 mm, which included an average error of 3.1 mm between in vivo and ex vivo MRI after applying DIR. The mean dice coefficient for the prostate contour between in vivo and ex vivo MRI increased from 0.83 before DIR to 0.93 after DIR.. The authors have developed a registration framework for mapping in vivo MRI data of the prostate with histology by implementing a number of processing steps and ex vivo MRI of the prostate specimen. Validation of DIR was challenging, particularly in prostates with few or mostly linear rather than spherical shaped features. Refinement of their MR imaging protocols to improve the data quality is currently underway which may improve registration accuracy. Additional mpMRI sequences will be registered within this framework to quantify prostate tumor location and biology.

Topics: Aged; Atlases as Topic; Cell Count; Fixatives; Formaldehyde; Gels; Histological Techniques; Humans; Imaging, Three-Dimensional; Magnetic Resonance Imaging; Male; Microtomy; Middle Aged; Prostate; Prostatectomy; Prostatic Neoplasms; Sepharose

To present a novel marker-flange, addressing source-reconstruction uncertainties due to the artifacts of a titanium intracavitary applicator used for magnetic resonance imaging (MRI)-guided high-dose-rate (HDR) brachytherapy (BT); and to evaluate 7 different MRI marker agents used for interstitial prostate BT and intracavitary gynecologic HDR BT when treatment plans are guided by MRI.. Seven MRI marker agents were analyzed: saline solution, Conray-60, copper sulfate (CuSO4) (1.5 g/L), liquid vitamin E, fish oil, 1% agarose gel (1 g agarose powder per 100 mL distilled water), and a cobalt-chloride complex contrast (C4) (CoCl2/glycine = 4:1). A plastic, ring-shaped marker-flange was designed and tested on both titanium and plastic applicators. Three separate phantoms were designed to test the marker-flange, interstitial catheters for prostate BT, and intracavitary catheters for gynecologic HDR BT. T1- and T2-weighted MRI were analyzed for all markers in each phantom and quantified as percentages compared with a 3% agarose gel background. The geometric accuracy of the MR signal for the marker-flange was measured using an MRI-CT fusion.. The CuSO4 and C4 markers on T1-weighted MRI and saline on T2-weighted MRI showed the highest signals. The marker-flange showed hyper-signals of >500% with CuSO4 and C4 on T1-weighted MRI and of >400% with saline on T2-weighted MRI on titanium applicators. On T1-weighted MRI, the MRI signal inaccuracies of marker-flanges were measured <2 mm, regardless of marker agents, and that of CuSO4 was 0.42 ± 0.14 mm.. The use of interstitial/intracavitary markers for MRI-guided prostate/gynecologic BT was observed to be feasible, providing accurate source pathway reconstruction. The novel marker-flange can produce extremely intense, accurate signals, demonstrating its feasibility for gynecologic HDR BT.

Topics: Brachytherapy; Catheters; Cobalt; Copper Sulfate; Feasibility Studies; Female; Fiducial Markers; Fish Oils; Humans; Iothalamate Meglumine; Magnetic Resonance Imaging, Interventional; Male; Phantoms, Imaging; Prostatic Neoplasms; Radiotherapy Dosage; Radiotherapy Planning, Computer-Assisted; Sepharose; Sodium Chloride; Titanium; Tomography, X-Ray Computed; Uncertainty; Uterine Cervical Neoplasms; Vitamin E

Carcinoma of the prostate is one of the most abundant killers for men in the western world, and it is frequently treated via Radiation therapy. Unfortunately, radiotherapy side effects include rectal irritation and bleeding, erectile dysfunction and urinary frequency. Because radiation intensity decays rapidly as a function of distance, displacing irradiated prostate away from normal tissues would reduce damage and therefore side effects. The objective of this study is to develop an inflatable balloon that is implanted via a minimal invasive procedure. The balloon is made of a biodegradable polymer called poly(lactide-co-epsilon-caprolactone). The implant is inserted rolled throughout the perineum; inflated in situ with a physiological saline; sealed and placed between the rectum wall, and the prostate gland. Balloon's mechanical and chemical properties were extensively characterized both in vitro and in vivo. The balloon's preparation ensures no bonding across surfaces as these may endanger the implant mechanical stability. Moreover, the coating method does not alter the polymer's molecular weight and therefore preserve its mechanical properties. Balloon's sterilization was carried out using ethylene oxide which, as our results show and in comparison with gamma-irradiation, doesn't damage the mechanical stability of the implant. The proper functionality of the insertion-mounting device as well as the balloon capability to retain its inflated form during patients' radiation session was demonstrated both in vitro and in vivo.

Topics: Absorbable Implants; Animals; Biocompatible Materials; Catheterization; Dogs; Gamma Rays; Guinea Pigs; Male; Materials Testing; Methylene Chloride; Molecular Weight; Prostatic Neoplasms; Radiation Protection; Radiotherapy; Sepharose; Sterilization

Imbalances in the epithelial-stromal interactions are important in the pathogenesis of prostate cancer. However, we know little about androgenic regulation in the stroma of prostate cancer. We examined the cancer-stromal interaction paying attention to androgen responsiveness of stromal side. In co-culture, PC3 and LNCaP cells did not affect dihydrotestosterone (DHT)-dependent growth of prostate stromal cells (PrSCs), but DU145 cells significantly reduced it. Conditioned medium from DU145 cells (DU145-CM) also inhibited DHT-dependent PrSCs growth, androgen receptor (AR) expression, and prostate specific antigen transcription. Although the inhibitory effect of DU145-CM was not affected by neutralizing antibody against EGF, FGF-2, or TNF-alpha, pretreatment with testosterone-Sepharose partially reduced the inhibitory ability of DU145-CM. These results suggest that DU145 cells produce inhibitory factors for androgen responsiveness, including steroid-binding protein(s), and these may participate in crosstalk between DU145 cells and PrSCs as modulators of androgen.

Topics: Carcinoma; Coculture Techniques; Culture Media, Conditioned; Dihydrotestosterone; Epidermal Growth Factor; Fibroblast Growth Factor 2; Humans; Male; Prostate; Prostate-Specific Antigen; Prostatic Neoplasms; Receptors, Androgen; Sepharose; Stromal Cells; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha

We determined the optimal conditions for the separation of N-glycosylation variants of prostate-specific antigen using concanavalin A. Concanavalin A is a lectin that binds to the terminal sugar residues of glycoproteins. We demonstrated that differences in the percentage of prostate-specific antigen bound to concanavalin A-Sepharose in patients with benign prostatic hyperplasia compared with patients with prostatic carcinoma, as described in the literature, arise when insufficient concanavalin A binding sites are added for complete binding of the glycosylation variants of prostate-specific antigen. We observed similar percentages of prostate-specific antigen bound to concanavalin A-Sepharose for benign prostatic hyperplasia (86.3% +/- 7.5, mean +/- SD) and carcinoma patients (81.8% +/- 12.0, mean +/- SD), when sufficient concanavalin A-Sepharose was added to allow optimal binding, and when samples with high prostate-specific antigen concentrations were not pre-diluted before incubation with concanavalin A-Sepharose. We conclude that differentiation of patients with benign prostatic hyperplasia or carcinoma of the prostate on the basis of differences in percentages of prostate-specific antigen bound to concanavalin A-Sepharose, i.e. separation of N-glycosylation variants, is not possible.

Topics: Binding Sites; Carcinoma; Chromatography, Affinity; Concanavalin A; Glycosylation; Humans; Male; Prostate-Specific Antigen; Prostatic Hyperplasia; Prostatic Neoplasms; Sepharose

Topics: Antigens, Neoplasm; Humans; Male; Prostate-Specific Antigen; Prostatic Hyperplasia; Prostatic Neoplasms; Sepharose

Topics: Acid Phosphatase; Antibody Specificity; Fluorescent Antibody Technique; Humans; Immunoglobulin G; Male; Neoplasms; Prostatic Neoplasms; Sepharose