Histology and histopathology Vol.31, nº3 (2016)
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- PublicationOpen AccessInfluence of BMP-2 on early follicular development and mRNA expression of oocyte specific genes in bovine preantral follicles cultured in vitro(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Ross, Rodrigo O.D.S.; da Cunha, Ellen V.; Portela, Antonia M.L.R.; Passos, José R.S.; Costa, José J.N.; Silva, Anderson W.B.; Saraiva, Márcia V.A.; Peixoto, Christina A.; Donato, Mariana A.M.; van den Hur, Robert k; Silva, José R.V.This study evaluates the effect of different concentrations (0, 10, 50 and 100ng/mL) of bone morphogenetic protein-2 (BMP-2) on primordial and secondary follicle development. It also investigates the effects of FSH and BMP-2 on the growth, morphology, ultrastructure and expression of mRNA for GDF9, NLRP5 and NPM2 genes in secondary follicles cultured for 18 days. The presence of BMP-2 at all tested concentrations increased the development of primordial follicles in vitro, but the highest concentration of BMP-2 (100 ng/mL) reduced the percentage of normal follicles when compared with tissues cultured with 10 ng/mL BMP-2. During culture of secondary follicles, in contrast to higher concentrations (50 or 100 ng/mL), 10 ng/mL BMP-2 kept the morphology of follicles during initial stages of in vitro culture. This concentration of BMP-2 also benefits maintenance of the ultrastructure of 18-day cultured follicles. The presence of both BMP-2 and FSH in culture medium resulted in a significant (P<0.05) increase in follicular diameter after 18 days of culture. However, both FSH and BMP-2 reduced follicular mRNA expression of GDF9 and NLRP5 when compared to follicles cultured in media containing only FSH. In combination with FSH, BMP-2 reduced the mRNA levels of NPM2, when compared to follicles cultured in control medium. It is concluded from these data that 10 ng/mL BMP-2 promotes the growth of primordial in vitro and it helps to maintain the ultrastructure of secondary follicles, while FSH is more important for better expression of follicular markers like GDF9 and NLRP5.
- PublicationOpen AccessImmunocytochemical localization of the calcium-binding proteins calbindin D28K, calretinin and parvalbumin in bat visual cortex(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Kim, Hang-Gu; Gu, Ya-Nan; Lee, Kyoung-Pil; Lee, Ji-Gun; Kim, Chan-Wook; Lee, Ji-Won; Jeong, Tae-Hee; Jeong, Young-Wun; Jeon, Chang-JinIt is a common misconception that bats are blind, and various studies have suggested that bats have visual abilities. The purpose of this study was to investigate the cytoarchitecture of calbindin D28K (CB)-, calretinin (CR)-, and parvalbumin (PV)-immunoreactive (IR) neurons in the bat visual cortex using immunocytochemistry. The highest density of CB- and PV-IR neurons was located in layer IV of the visual cortex. The majority of CB- and PV-IR neurons were characterized by a stellate or round/oval shape. CR-IR neurons were predominantly located in layers II/III, and the cells were principally round/oval in shape. Two-color immunofluorescence revealed that 65.96%, 24.24%, and 77.00% of the CB-, CR-, and PV-IR neurons, respectively, contained gamma-aminobutyric acid (GABA). We observed calcium-binding protein (CBP)- IR neurons in specific layers of the bat visual cortex and in specific cell types. Many of the CBP-IR neurons were GABAergic interneurons. These data provide useful clues to aid in understanding the functional aspects of the bat visual system.
- PublicationOpen AccessGLI1 expression is an important prognostic factor that contributes to the poor prognosis of rhabdomyosarcoma(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Wang, Yuanyuan; Sun, Chao; Jiang, Jinfang; Xie, Yuwen; Li, Bingcheng; Cui, Xiaobin; Chen, Yunzhao; Liu, Chunxia; Li, FengThe GLI1 and MDM2 genes are amplified or exhibit copy number gains in rhabdomyosarcoma (RMS). Here, we used immunohistochemistry to determine the relationships between GLI1 and MDM2 protein expression and several clinicopathological variables of RMS. GLI1 and MDM2-positivity rates were 61.36% and 13.64%, respectively. GLI1 expression correlated with presence of the PAX3-FOXO1 fusion gene (P=0.040) and lymph node metastasis (P=0.034), and a significant association was found between GLI1 expression and overall survival (OS) (P=0.008). However, there was no association between MDM2 expression and any of the clinicopathological parameters or OS. Thus, GLI1 may be a biomarker of poor prognosis in RMS patients, and could itself be a therapeutic target. This contrasts with the apparent lack of clinical importance of MDM2 in RMS pathology, at least in the cohorts we examined.
- PublicationOpen AccessDiagnostic value of matrix metalloproteinase 9 and tissue inhibitor of matrix metalloproteinases 1 in cholesteatoma(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Olszewska, Ewa; Matulka, Marlena; Mroczko, Barbara; Pryczynicz, AnnaObjectives: Matrix metalloproteinase 9 (MMP-9), able to degrade type IV collagen, plays a key role in inflammatory cell migration as well as in the destructive behaviour of cholesteatoma. The aim of our study was to compare the expression of MMP-9 and TIMP-1 in cholesteatoma tissue and in the concentrations in serum and plasma concentrations. Material and Methods: Twenty five adult patients suffering from cholesteatoma (a study group) were included in the study. A comparison group consisted of 25 adult patients admitted to hospital due to nasal septum deviation. MM-9 and TIMP-1 serum and plasma concentrations as well as proteins’ expressions in cholesteatoma tissues (study group) and normal retroauricular skin specimens (control group) were evaluated. MMP-9 and TIMP-1 concentrations were measured using enzyme-linked immunosorbent assay (ELISA). Cholesteatoma tissues and normal retroauricular skin specimens were evaluated immunohistochemically. Results: In the study and comparison groups, MMP9 and TIMP-1 concentrations were similar with no significant difference within the groups. In cholesteatoma tissues, the expression of the investigated enzyme and its inhibitor was higher than in normal skin specimens, limited mostly to cholesteatoma perimatrix. Conclusion: Cholesteatoma may be limited to the middle ear or parts of the temporal bones. Our findings suggest better clinical usefulness of MMP-9 and TIMP-1 expression in cholesteatoma tissues than either serum or plasma levels of these proteins. It might suggest that the higher the expression of MMP-9 the stronger the inflammation -accompanied cholesteatoma.
- PublicationOpen AccessClinical significance of stem cell marker CD133 expression in colorectal cancer(Universidad de Murcia. Departamento de Biología Celular e Histología, 2016) Wang, Bin-Bin; Li, Zhi-juan; Zhang, Feng-Feng; Hou, Hai-Tao; Yu, Jing-Kui; Li, FengObjective: CD133, a glycoprotein, is expressed in different types of human stem cells and tumor cells. Detection of altered CD133 expression in colorectal cancer tissues could be useful as a marker for the prediction of colorectal tumorigenesis, progression, and prognosis. Methods: A total of 19 fresh and 145 paraffin-embedded tissue specimens from colorectal cancer patients were obtained for detection of CD133 expression using flow cytometry and immunohistochemistry, respectively. The tumorigenic capacity of tumor cells from 19 patients was assessed in nude mice. Association of CD133 expression was then analyzed for clinical significance. Results: The percentage of CD133- positive (CD133+) tumor cell population ranged between 0.84% and16.75% (mean ratio=7.15%) of tumor cells in the 19 freshly isolated tissue samples. CD133 expression in tumor cells was associated with tumor lymph node metastasis (9.81% vs. 3.22%; p=0.013) and poor tumor differentiation (8.32% vs. 5.07%; p=0.043). In the 145 paraffin-embedded samples, CD133+ colorectal cancer was also associated with local recurrence of tumorigenesis (p=0.035) and distant metastasis (p=0.017), while patients with over 5% CD133+ tumor cells exhibited a decreased survival rate (p=0.001). Multivariate COX analysis showed that the depth of tumor invasion, histology, stages, lymph node metastasis, and CD133 expression were all independent prognosis factors for colorectal cancer (p=0.032, 0.011, 0.001, 0.002, and 0.030, respectively). Furthermore, as few as 5,000 CD133+ colorectal cancer HCT116 cellswere sufficient to form tumor xenografts, whereas 1x105 CD133- tumor cells failed to develop tumor xenografts in nude mice. Conclusions: CD133 expression is a useful biomarker for prediction of colorectal cancer progression and survival of patients.
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