Isolation and characterization of Wharton's jelly-derived multipotent mesenchymal stromal cells obtained from bovine umbilical cord and maintained in a defined serum-free three-dimensional system

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Results of prostate cancer screening in non-symptomatic men

Objectives: To verify prostate cancer prevalence in non-symptomatic men between 50 and 70 years old as well as cancer characteristics. Material and Methods: 2815 non-symptomatic men had total PSA and digital rectal examination performed between March 1998 and April 1998. Racial distribution was: 2331 Caucasians (83.9%), 373 Blacks (13.4%) and 75 Asiatic (2.7%). PSA was normal in 2554 (91.4%), 4 to 10 in 177 (6.3%) and greater than 10 in 64 (2.3%). DRE was normal in 2419 (86.3%), suspicious in 347 (12.4%) and characteristic for cancer in 37 (1.3%). Men with abnormal DRE and/or PSA had transrectal prostate biopsy indicated. Results: 461 biopsies were done and 78 tumors was detected (prevalence = 2.8%). Prevalence was progressively higher with age (p < 0.001), PSA level (p < 0.0001) and DRE findings (p = 0.0216). Cancer prevalence in Blacks was 1.65 times higher than in Caucasians (p > 0.05) and 94.9% of detected tumors were moderately or poorly differentiated. Sensibility, specificity, positive predictive value, negative predictive value and total accuracy for PSA were respectively: 66.6%; 89.7%; 51.7%; 94.2% and 86.5%. For DRE, the respective values were: 49.1%; 79.4%; 50.9%; 78.3% and 70.3%. Conclusions: prostate cancer prevalence in the studied population (2.8%) was similar to that of other countries populations. Cancer prevalence in blacks was 1.65 times higher than in Caucasians (difference was not statistically significant). Cancer prevalence becomes higher with aging. The association of DRE and PSA is of paramount importance for cancer diagnosis. The great majority of detected tumors (94.9%) was moderately and poorly differentiated. Brazil probably needs regional studies to better characterize prostate cancer epidemiology due to population heterogeneity.

Single testicular metastasis mimicking primary testicular neoplasm: A rare manifestation of prostate cancer

The incidence of secondary testicular tumors ranges from 0.02 to 2.5% among autopsies in general. With the exception of leukemias and lymphomas, prostate cancer is the most common primary site. It is diagnosed in autopsies or incidentally, following therapeutic orchiectomies in more advanced stages of the disease. In the present report, we show a case of testicular metastasis derived from prostate neoplasm whose clinical presentation as a single metastasis was similar to a primary testicular neoplasm. The diagnosis was evidenced after orchiectomy by histological examination and immunohistochemical tests.

Lack of reliability of nanotechnology in the of free plasma DNA in samples of patients with prostate cancer

Background: Several studies seek biological markers that give diagnostic and degree of tumor development. The aim of this study was to validate the determination of plasma DNA using nanotechnology (Nanovue™-NV) in samples of 80 patients with prostate cancer. Methods. Blood samples of 80 patients of the Urology Ambulatory of Faculdade de Medicina do ABC with prostate cancer confirmed by anatomical-pathology criteria were analyzed. DNA extraction was performed using a GFX TM kit (Amersham Pharmacia Biotech, Inc, USA) following the adapted protocol. Plasma was subjected to centrifugation. Results: There was a big difference between the first and the second value obtained by NanoVue Only two samples had no differences between duplicates. Maximum difference between duplicates was 38 μg/mL. Average variation between 51 samples was 10.29 μg/mL, although 21 samples had differences above this average. No correlation was observed between pDNA obtained by traditional spectrophotometry and by nanotechnology. Conclusion: Determination of plasma DNA by nanotechnology was not reproducible. © 2013 Moreno et al; licensee BioMed Central Ltd.

Isolation and characterization of equine peripheral blood-derived multipotent mesenchymal stromal cells

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Multi-scale lacunarity as an alternative to quantify and diagnose the behavior of prostate cancer

Prostate cancer is a serious public health problem accounting for up to 30% of clinical tumors in men. The diagnosis of this disease is made with clinical, laboratorial and radiological exams, which may indicate the need for transrectal biopsy. Prostate biopsies are discerningly evaluated by pathologists in an attempt to determine the most appropriate conduct. This paper presents a set of techniques for identifying and quantifying regions of interest in prostatic images. Analyses were performed using multi-scale lacunarity and distinct classification methods: decision tree, support vector machine and polynomial classifier. The performance evaluation measures were based on area under the receiver operating characteristic curve (AUC). The most appropriate region for distinguishing the different tissues (normal, hyperplastic and neoplasic) was defined: the corresponding lacunarity values and a rule's model were obtained considering combinations commonly explored by specialists in clinical practice. The best discriminative values (AUC) were 0.906, 0.891 and 0.859 between neoplasic versus normal, neoplasic versus hyperplastic and hyperplastic versus normal groups, respectively. The proposed protocol offers the advantage of making the findings comprehensible to pathologists. (C) 2014 Elsevier Ltd. All rights reserved.

Relevance of the Myeloid Differentiation Factor 88 (MyD88) on RANKL, OPG, and Nod Expressions Induced by TLR and IL-1R Signaling in Bone Marrow Stromal Cells

The myeloid differentiation factor 88 (MyD88) plays a pivotal role in Toll-like receptor (TLR)- and interleukin-1 receptor (IL-1R)-induced osteoclastogenesis. We examined the role of MyD88 on p38 mitogen-activated protein kinase (MAPK) and nuclear factor kappa-light-chain-enhancer of activated B cell (NF-κB) activation and nucleotide-binding oligomerization domain (Nod) induction by lipopolysaccharide (LPS) and IL-1 beta, and their effect on receptor activator of NF-κB ligand (RANKL) and osteoprotegerin (OPG) production in bone marrow stromal cell (BMSC). RANKL, Nod1, Nod2, NF-κB, and p38 protein levels were determined by Western blot. Nod2 was stimulated with muramyl dipeptide (MDP) prior to TLR4 stimulation with LPS. MyD88 deficiency markedly inhibited RANKL expression after LPS stimulation and increased OPG messenger RNA (mRNA) production. Also, MyD88 was necessary for NF-κB and p38 MAPK activation. MDP alone did not induce RANKL and OPG expressions; however, when combined with LPS, their expressions were significantly increased (p < 0.05). Our results support that MyD88 signaling has a pivotal role in osteoclastogenesis thought NF-κB and p38 activation. Nod2 and especially Nod1 levels were influenced by MyD88.

Combining xanthan and chitosan membranes to multipotent mesenchymal stromal cells as bioactive dressings for dermo-epidermal wounds

The association between tridimensional scaffolds to cells of interest has provided excellent perspectives for obtaining viable complex tissues in vitro, such as skin, resulting in impressive advances in the field of tissue engineering applied to regenerative therapies. The use of multipotent mesenchymal stromal cells in the treatment of dermo-epidermal wounds is particularly promising due to several relevant properties of these cells, such as high capacity of proliferation in culture, potential of differentiation in multiple skin cell types, important paracrine and immunomodulatory effects, among others. Membranes of chitosan complexed with xanthan may be potentially useful as scaffolds for multipotent mesenchymal stromal cells, given that they present suitable physico-chemical characteristics and have adequate tridimensional structure for the adhesion, growth, and maintenance of cell function. Therefore, the purpose of this work was to assess the applicability of bioactive dressings associating dense and porous chitosan-xanthan membranes to multipotent mesenchymal stromal cells for the treatment of skin wounds. The membranes showed to be non-mutagenic and allowed efficient adhesion and proliferation of the mesenchymal stromal cells in vitro. In vivo assays performed with mesenchymal stromal cells grown on the surface of the dense membranes showed acceleration of wound healing in Wistar rats, thus indicating that the use of this cell-scaffold association for tissue engineering purposes is feasible and attractive.

Relevance of the Myeloid Differentiation Factor 88 (MyD88) on RANKL, OPG, and Nod Expressions Induced by TLR and IL-1R Signaling in Bone Marrow Stromal Cells

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Combining xanthan and chitosan membranes to multipotent mesenchymal stromal cells as bioactive dressings for dermo-epidermal wounds

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Combining xanthan and chitosan membranes to multipotent mesenchymal stromal cells as bioactive dressings for dermo-epidermal wounds

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

High fat-induced obesity associated with insulin-resistance increases FGF-2 content and causes stromal hyperplasia in rat ventral prostate

Obesity affects sex hormone secretion, which can negatively influence prostatic structure, homeostasis, and disease. This investigation aimed to evaluate the repercussions of obesity induced by a high-fat diet on the rat prostate, with or without treatment with the aromatase inhibitor, Letrozole. Adult Wistar rats were fed a high-fat diet (20% saturated fat, O) for 15 weeks to induce obesity or received a balanced diet (4% fat, C). Then, a group of C and O rats were daily treated with Letrozole (1 mg/kg b.w. per day) for 2 weeks (CL and OL, respectively). Subsequently, ventral prostate was processed for analysis by transmission electron microscopy, immunohistochemistry, and Western blotting. Obesity decreased 70% of the testosterone plasma level. The prostate showed epithelial atrophy and dilated acini in the intermediate portion and epithelial wrinkling in the distal tips. The relative frequency of smooth muscle alpha-actin in the O group increased by 67%. Ultrastructurally, epithelial cells in obese animals presented altered secretory organelles, lipid droplets, and thicker subjacent fibromuscular layer. Letrozole treatment caused a partial restoration of the prostatic changes caused by obesity. Obesity increased the prostatic content of fibroblast growth factor-2 (FGF-2) by 150%, and Letrozole treatment increased this protein even more in the control and obese groups. This investigation shows that obesity provokes structural and ultrastructural changes in the epithelium of rat prostate; these changes might affect gland homeostasis and physiology. The epithelial and smooth muscle cell hyperplasia and increased FGF-2 expression observed in this experimental model of obesity/insulin-resistance might explain the high frequency of benign prostatic hyperplasia in insulin-resistant men.

Prostate carcinogenesis induced by N-methyl-N-nitrosourea (mnu) in gerbils: Histopathological diagnosis and potential invasiveness mediated by extracellular matrix components

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Anatomy of smooth muscle cells in nonmalignant and malignant human prostate tissue

Differently graded areas of human prostate adenocarcinoma were examined after Masson's trichrome staining or immunohistochemistry for smooth muscle alpha-actin, type IV collagen and laminin. In addition, the ultrastructure of the prostatic smooth muscle cells (SMC) during glandular proliferation and epithelial invasion in selected tumors was studied. The SMC formed a thick layer below the epithelial structures in unaffected areas and were closely associated with each other in homotypic interactions. As the tumor grade increased, the SMC gradually lost interactions with each other and became atrophic. With the growth of the epithelial compartment, the SMC initially segregated to the tumor periphery and the intercellular spaces increased. In high grade tumors, the epithelial cancer cells invaded the spaces between the SMC. Immunohistochemical analysis of the basal membrane revealed increased disruption of the usually thick basal membrane, which became thinner and faintly stained with each of the antibodies used. We conclude that most SMC become atrophic following epithelial invasion in human tumors and that degradation of the basal membrane is an important factor in this process. At the ultrastructural level, different SMC phenotypes occur in prostatic tissues during epithelial invasion. Interconversion between these phenotypes is suggested and a probable relationship among them is proposed.