Patrick Vianna Garcia

Morphometric-stereological and functional epididymal alterations and a decrease in fertility in rats treated with finasteride and after a 30-day post-treatment recovery period

OBJECTIVE To evaluate morphometric-stereological changes in the epididymal caput, sperm quality, and fertility parameters in rats treated with finasteride and after a 30-day post-treatment recovery period. DESIGN Experimental study in a research laboratory. SETTING Reproductive biology research laboratory. ANIMAL(S) Male and female Sprague Dawley rats. INTERVENTION(S) Treatment with finasteride (5 mg/kg/day) for 56 days followed by 30 days without treatment. MAIN OUTCOME MEASURE(S) Serum hormone analyses, morphometric-stereological and ultrastructural evaluation of the epididymal caput, sperm transit time, natural mating, in utero insemination, sperm membrane integrity, and fertility parameters. RESULT(S) Serum dihydrotestosterone levels in the finasteride group decreased by ~40% compared with that of control rats. Ultrastructural analysis revealed significant reductions in several morphometric-stereological parameters of the epididymal caput. All parameters recovered significantly in the post-treatment period. There was no alteration in daily sperm production in the finasteride group. However, significant reductions in sperm transit time, motility, sperm membrane integrity, and fertility parameters were observed in rats treated with finasteride. CONCLUSION(S) Treatment with finasteride caused morphometric-stereological and functional changes in the epididymis and in sperm function that led to a reduction in fertility parameters. A 30-day post-treatment recovery period was insufficient to restore normal sperm motility, sperm transit time, and some fertility parameters.

The Epididymis: Embryology, Structure, Function and Its Role in Fertilization and Infertility

The anatomic segments of the epididymis include the initial segment, the caput, the corpus and the cauda. Each region consists of a lumen and a polarized epithelium composed mostly of principal and basal cells (Lasserre et al., 2001; Dacheux et al., 2005). Although these four anatomical regions of the epididymis are easily identified in most adult male mammals (Yanagimachi et al., 1985; Smithwick & Young, 2001), histological and ultrastructural segmentation of this organ varies among the different phylogenies of mammals. The rat epididymis is most commonly adopted as an experimental model of study (Figure 1).

Characterization of reactive stroma in prostate cancer: involvement of growth factors, metalloproteinase matrix, sexual hormones receptors and prostatic stem cells

ABSTRACT Introduction and Objectives: Reactive Stroma (RStr) is observed in many human cancers and is related to carcinogenesis. The objectives of the present study were to stablish a relationship of the RStr microenvironment with prostate cancer (Pca) through a morphological and molecular characterization, and to identify a possible relationship between RStr with worse prognosis factors and occurrence of malignant prostatic stem cells. Materials and Methods: Forty prostatic samples were selected from men with Pca diagnosis submitted to radical prostatectomy; they were divided in two groups: Group-1 (n=20): samples without reactive stroma; Group-2 (n=20): samples of PCa with intense stroma reaction. Prostatic samples were evaluated for RStr intensity by Masson Trichromic stain and posteriorly submitted to histopathological and immunohistochemistry analysis for antigens: α-actin, vimentin, IGF-1, MMP-2, FGF-2, C-Myc, PSCA, AR, Erα and ERβ. Results: Reactive stroma with intense desmoplastic reactivity was significantly more frequent in intermediate (Gleason 7, 3+4) and high grade tumors (Gleason 7, 4+3). The group with intense stromal reactivity showed significant higher levels of Vimentin, IGF-1, MMP-2, FGF-2, C-Myc, PSCA and ERα. Conclusions: It can be concluded that RStr may be a predictive marker of Pca progression, since it was associated with increase of growth factors, imbalance of androgen and estrogen receptors and presence of malign prostatic stem cells.

Orchidopexy restores morphometric-stereologic changes in the caput epididymis and daily sperm production in cryptorchidic mice, although sperm transit time and fertility parameters remain impaired

OBJECTIVE To evaluate the changes in the caput epididymis following cryptorchidism and orchidopexy. DESIGN Experimental study in a research laboratory. SETTING Reproductive biology research laboratory. ANIMAL(S) Immature male and mature female mice (C57BL/6). INTERVENTION(S) Experimental cryptorchidism and orchidopexy. MAIN OUTCOME MEASURE(S) Morphometric-stereologic analyses, serum testosterone dosage, immunohistochemical staining of the antigen TRA54 (testicular germ cells immunized to a rat monoclonal antibody), smooth muscle α-actin (SM α-actin) and SM myosin heavy chain, sperm transit time, and fertility parameters. RESULT(S) There was a significant reduction in the morphometric-stereologic parameters in the cryptorchidic mice. These parameters demonstrated significant recovery following orchidopexy. Staining for an androgen-dependent antigen, TRA54, was observed in all groups. SM α-actin and SM myosin heavy chain staining was significantly increased in the cryptorchidism group but stable in the orchidopexy group. Despite the recovery of daily sperm production in the testes, the sperm transit time in the epididymis and fertility parameters remained significantly reduced in the orchidopexy group. CONCLUSION(S) In cryptorchidic animals, there was an acceleration of sperm passage through the epididymal duct. Orchidopexy did not restore the normal passage time. Accordingly, there was a significant reduction in the fertility parameters in the cryptorchidic group that were not fully recovered following orchidopexy.

Potential therapeutic strategies for non - muscle invasive bladder cancer based on association of intravesical immunotherapy with P-MAPA and systemic administration of cisplatin and doxorubicin

ABSTRACT The present study describes the histopathological and molecular effects of P-MAPA (Protein aggregate magnesium-ammonium phospholinoleate-palmitoleate anhydride) intravesical immunotherapy combined with systemic doxorubicin or cisplatin for treatment of non-muscle invasive bladder cancer (NMIBC) in an appropriate animal model. Our results showed an undifferentiated tumor, characterizing a tumor invading mucosa or submucosa of the bladder wall (pT1) and papillary carcinoma in situ (pTa) in the Cancer group. The histopathological changes were similar between the combined treatment with intravesical P-MAPA plus systemic Cisplatin and P-MAPA immunotherapy alone, showing decrease of urothelial neoplastic lesions progression and histopathological recovery in 80% of the animals. The animals treated systemically with cisplatin or doxorubicin singly, showed 100% of malignant lesions in the urinary bladder. Furthemore, the combined treatment with P-MAPA and Doxorubicin showed no decrease of urothelial neoplastic lesions progression and histopathological recovery. Furthermore, Akt, PI3K, NF-kB and VEGF protein levels were significantly lower in intravesical P-MAPA plus systemic cisplatin and in intravesical P-MAPA alone treatments than other groups. In contrast, PTEN protein levels were significantly higher in intravesical P-MAPA plus systemic cisplatin and in intravesical P-MAPA alone treatments. Thus, it could be concluded that combination of intravesical P-MAPA immunotherapy and systemic cisplatin in the NMIBC animal model was effective, well tolerated and showed no apparent signs of antagonism between the drugs. In addition, intravesical P-MAPA immunotherapy may be considered as a valuable option for treatment of BCG unresponsive patients that unmet the criteria for early cystectomy.

Increased toll-like receptors and p53 levels regulate apoptosis and angiogenesis in non-muscle invasive bladder cancer: mechanism of action of P-MAPA biological response modifier

BackgroundThe new modalities for treating patients with non-muscle invasive bladder cancer (NMIBC) for whom BCG (Bacillus Calmette-Guerin) has failed or is contraindicated are recently increasing due to the development of new drugs. Although agents like mitomycin C and BCG are routinely used, there is a need for more potent and/or less-toxic agents. In this scenario, a new perspective is represented by P-MAPA (Protein Aggregate Magnesium-Ammonium Phospholinoleate-Palmitoleate Anhydride), developed by Farmabrasilis (non-profit research network). This study detailed and characterized the mechanisms of action of P-MAPA based on activation of mediators of Toll-like Receptors (TLRs) 2 and 4 signaling pathways and p53 in regulating angiogenesis and apoptosis in an animal model of NMIBC, as well as, compared these mechanisms with BCG treatment.ResultsOur results demonstrated the activation of the immune system by BCG (MyD88-dependent pathway) resulted in increased inflammatory cytokines. However, P-MAPA intravesical immunotherapy led to distinct activation of TLRs 2 and 4-mediated innate immune system, resulting in increased interferons signaling pathway (TRIF-dependent pathway), which was more effective in the NMIBC treatment. Interferon signaling pathway activation induced by P-MAPA led to increase of iNOS protein levels, resulting in apoptosis and histopathological recovery. Additionally, P-MAPA immunotherapy increased wild-type p53 protein levels. The increased wild-type p53 protein levels were fundamental to NO-induced apoptosis and the up-regulation of BAX. Furthermore, interferon signaling pathway induction and increased p53 protein levels by P-MAPA led to important antitumor effects, not only suppressing abnormal cell proliferation, but also by preventing continuous expansion of tumor mass through suppression of angiogenesis, which was characterized by decreased VEGF and increased endostatin protein levels.ConclusionsThus, P-MAPA immunotherapy could be considered an important therapeutic strategy for NMIBC, as well as, opens a new perspective for treatment of patients that are refractory or resistant to BCG intravesical therapy.

Energetic Metabolism in the Treatment of Prostate Cancer: Effects of P-MAPA Immunomodulator

The American Cancer Society estimates that 161.360 new cases of Prostate Cancer will be diagnosed in the United States of America (USA) in 2017. Faced with such alarming data, research into new terapeutic options is necessary, and the P-MAPA immunomodulator opens a new perspective for the combat of this type of cancer. Therefore, the aim of this study is to characterize the molecular effects of P-MAPA immunotherapy in Prostate Cancer treatment involving cellular energy metabolism.

Steroid Hormone Receptors as Potential Mediators of the Clinical Effects of Dutasteride: A Prospective, Randomized, Double-Blind Study.

This study characterizes the clinical and morphofunctional effects of a 5α-reductase inhibitor on steroid hormone receptors in normal human prostate tissue, as potential mediators of the clinical effects of dutasteride. This work was a prospective, double-blind, and randomized study that evaluated 49 men aged between 45 and 70 years, with no alterations in a digital rectal examination and prostate-specific antigen measurements between 2.5 and 4.0 ng/mL. These patients underwent prostate biopsy guided by transretal ultrasound with prostate neoplasia being ruled out, and the patients were divided into two groups, with one group receiving dutasteride (n = 25) and one group receiving a placebo (n = 24). The patients were clinically assessed each quarter, and at the end of 12 months they underwent new laboratory tests, prostate rebiopsy, and histopathological, immunohistochemical and clinical analyses. The estrogen receptor-beta (ERβ) and androgen receptor immunoreactivities were higher, and the proliferation/apoptotic ratio was significantly lower with predominance of the apoptotic process, followed by a significant reduction in the prostate volume and the total serum prostate-specific antigen levels in the dutasteride group when compared with the placebo group, with a clear supremacy of ERβ. There were no significant variations in the serum estrogen and testosterone levels, in the body mass index, or in the ERα immunoreactivities in the dutasteride and placebo groups. The results demonstrated the importance of the ERβ pathway in the activation mechanisms of apoptosis, exerting a protective effect in the normal prostate, indicating that this receptor might be an important mediator of the clinical effects of dutasteride.

EFFECTS OF P-MAPA ASSOCIATED WITH GEMCITABINE IN THE PANCREATIC CANCER: HISTOLOGIC EFFECTS

Currently antitumor therapies consist in the neoplasic cells attack by chemotherapy and radiotherapy. Successful treatment depends on the type of cancer, because each cancer has particular characteristics of survival and immune evasion. The neoplasic cell biology studies demonstrated a persistent immunosuppressive environment surrounding transformed cells. Thus, compounds that are capable of acting as agonists of Toll-like receptors (TLRs) may represent promising candidates to be developed as medicaments against cancer. Thus, the general objectives of this study are to characterize and compare the histologic effects of P-MAPA associated with the chemotherapeutic agent Gemcitabine to treat pancreatic cancer in rats.

Steroid Hormone Receptors as Potential Mediators of the Clinical Effects of Dutasteride

This study characterizes the clinical and morphofunctional effects of a 5α-reductase inhibitor on steroid hormone receptors in normal human prostate tissue, as potential mediators of the clinical effects of dutasteride. This work was a prospective, double-blind, and randomized study that evaluated 49 men aged between 45 and 70 years, with no alterations in a digital rectal examination and prostate-specific antigen measurements between 2.5 and 4.0 ng/mL. These patients underwent prostate biopsy guided by transretal ultrasound with prostate neoplasia being ruled out, and the patients were divided into two groups, with one group receiving dutasteride (n = 25) and one group receiving a placebo (n = 24). The patients were clinically assessed each quarter, and at the end of 12 months they underwent new laboratory tests, prostate rebiopsy, and histopathological, immunohistochemical and clinical analyses. The estrogen receptor-beta (ERβ) and androgen receptor immunoreactivities were higher, and the proliferation/apoptotic ratio was significantly lower with predominance of the apoptotic process, followed by a significant reduction in the prostate volume and the total serum prostate-specific antigen levels in the dutasteride group when compared with the placebo group, with a clear supremacy of ERβ. There were no significant variations in the serum estrogen and testosterone levels, in the body mass index, or in the ERα immunoreactivities in the dutasteride and placebo groups. The results demonstrated the importance of the ERβ pathway in the activation mechanisms of apoptosis, exerting a protective effect in the normal prostate, indicating that this receptor might be an important mediator of the clinical effects of dutasteride.