Histology and histopathology Vol.29, nº 3 (2014)

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  • Publication
    Open Access
    Metallothioneins and trace elements dyshomeostasis induced by exposure to gasoline vapor in mice
    (F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2014) Grebić, Damir; Tota, Marin; Jakovac, Hrvoje; Broznić, Dalibor; Marinić, Marinić; Canadi, Gordana; Milin, Čedomila; Radošević-Stašić, Biserka
    To investigate the effects of air pollution related with the gasoline/petrochemical industry the expression of metallothionein I (MT-I) mRNA and tissue metals were analyzed in organs of mice, exposed to gasoline (G) vapor in laboratory conditions. Control groups consisted of intact mice and of those exposed in the metabolic chamber to fresh air. The data obtained by RT-PCR and inductively coupled plasma spectrometry have shown that exposure to G vapor leads to upregulation of MT-I mRNA in organs that receive a strong respiratory and olfactory input or participate in gasoline degradation and elimination (lungs, brain, kidney and liver). Besides, in the brain and in the lungs, kidney and liver a decreased tissue content of Zn2+ or Cu2+ and Mg2+ was found (p<0.001). Some of these changes were obtained also in mice closed in the metabolic chamber, pointing to the involvement of stress-induced mechanisms in the transcriptional regulation of MTs
  • Publication
    Open Access
    Eosinophil depletion protects mice from tongue squamous cell carcinoma induced by 4-nitroquinoline-1-oxide
    (F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2014) Silva, Janine Mayra da; Queiroz-Junior, Celso Martins; Batista, Aline Carvalho; Rachid, Milene Alvarenga; Teixeira, Mauro Martins; Silva, Tarcília Aparecida da
    Aims: Tumor-associated tissue eosinophilia (TATE) has been correlated with prognosis in oral squamous cell carcinoma (OSCC). This study aimed to investigate whether eosinophils depletion affects experimental oral carcinogenesis. Methods and Results: BALB/c (wild type - WT) and eosinophil-deficient (Δdb/GATA-1) mice were treated with the carcinogen 4-nitroquinoline-1-oxide (4NQO) in drinking water for 28 weeks. Tongues were collected for histopathological and immunohistochemical analysis, as well as for the evaluation of cytokines/chemokines by ELISA. The tongue SCC induced by 4NQO was associated with a rise in eosinophil numbers. WT-treated group showed a significantly increased incidence of SCC, with higher cytological atypia, in comparison with Δdb/GATA-1 mice. Consistently, the proliferative index was higher in WT compared to the Δdb/GATA-1/GATA1-treated group. No significant changes in the concentration of CCL3, CCL11 and TNF-α were detected for both groups after 4NQO treatment. Conclusions: These results suggest that eosinophils might be responsible for the deleterious outcome of experimental tongue carcinogenesis, given that their ablation protects mice from OSCC.
  • Publication
    Open Access
    Cooperative role between p21cip1/waf1 and p27kip1 in premature senescence in glandular proliferative lesions in mice
    (F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2014) García-Fernández, R.A.; García-Palencia, P.; Suárez, C.; Sánchez, M.A.; Gil-Gómez, G.; Sánchez, B.; Rollán, E.; Martín-Caballero, J.; Flores, J.M.
    Cellular senescence has been considered a novel target for cancer therapy. It has also been pointed out that p21cip1/waf1 and p27kip1 cyclin-dependent kinase inhibitors (CKIs) play a role in cellular senescence in some tumor types. Therefore, in order to address the possibility of a cooperative role between p21 and p27 proteins in senescence in vivo we analyzed cellular senescence in spontaneous glandular proliferative lesions (adrenal, thyroid and pituitary glands) in a double-KO mice model, using γH2AX, p53, p16, PTEN and Ki67 as senescence markers. The results obtained showed that p21p27 double-null mice had the lowest number of γH2AX positive cells in glandular hyperplasias and benign tumors. Also, in this group, Ki67 proliferation index correlated with a lower immunohistochemical expression of γH2AX and p53. The expression of p16 and PTEN do not seem to cause synergism of senescence in the benign lesions analyzed in p21p27 double-KO mice. These observations suggest an intrinsic cooperation between p21 and p27 CKIs in the activation of stress-induced cellular senescence and tumor progression in vivo, which would be a physiological mechanism to prevent tumor cell proliferation.
  • Publication
    Open Access
    Proteomic analysis in usual and nonspecific interstitial pneumonia
    (F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2014) Ohara, Ichiyo; Aida, Shinsuke; Shimazaki, Hideyuki; Kobayashi, Hideo; Tsuda, Hitoshi; Toda, Tosifusa; Nakanishi, Kuniaki; Tamai, Seiichi
    Differentiating nonspecific interstitial pneumonia (NSIP) from usual interstitial pneumonia (UIP) is important for the determination of both treatment and prognosis. Using two-dimensional fluorescence difference gel electrophoresis (2D-DIGE), we examined 8 UIPs, 8 NSIPs, and 30 normal lung tissues. Comparisons with control in 2D-DIGE showed that (a) in UIP, nine protein spots were significantly upregulated and seven were significantly downregulated, (b) in NSIP, four protein spots were significantly upregulated and nine were significantly downregulated. The detected proteins were analyzed by MALDI-TOF mass spectrometry, allowing qualitative differences in vimentin subtypes to be characterized. One vimentin subtype was upregulated in UIP, while another one was downregulated in NSIP (vs. control). These different characteristics were partially supported by the results of Western blot analysis. Our immunohistochemistry revealed vimentin expression within fibroblasts (a) in fibroblastic foci in UIP and (b) in fibrotic alveolar walls in NSIP. Differences in vimentin subtypes may provide useful biomarkers for separating NSIP from UIP, alongside differences in histological characteristics.
  • Publication
    Open Access
    Remote lung injury after experimental intestinal ischemia-reperfusion in horses
    (F. Hernández y Juan F. Madrid. Universidad de Murcia. Departamento de Biología Celular e Histología, 2014) Montgomery, Julia B.; Hamblin, Brie; Suri, Sarabjeet Singh; Johnson, Laura E.; New, Dallas; Johnston, Jennifer; Kelly, Jenny; Wilson, David G.; Singh, Baljit
    Ischemia followed by reperfusion leads to release of toxic molecules into the circulation, and these molecules may cause injury in remote organs such as the lung. Horses commonly suffer from episodes of intestinal ischemia-reperfusion (IR) due to intestinal twisting/strangulation followed by repair. Because there is no evidence of lung injury associated with IR in horses, we designed a study to characterize the intestinal IR-associated lung inflammation and determine the effect of lidocaine on lung inflammation in IR horses. Lung tissues were collected from non-anesthetized (n=4) and anesthetized (n=4) control horses and horses (n=12) after 70 minutes of ischemia followed by 60 minutes of reperfusion. Horses in IR groups received Lactated Ringer’s Solution (LRS; n=6) or lidocaine (n=6) intravenously. Control lungs had normal histology but lungs from IR horses showed moderate accumulation of neutrophils in blood vessels and airways. We found increased staining for TLR4, IL-8, TLR9, and von Willebrand factor (vWF) along with aggregates of vWFpositive platelets in lung vessels of IR horses compared to the controls. Lung TNFα was significantly increased in IR horses compared to the control horses (P<0.05). Neutrophil numbers, but not MPO concentrations, were significantly lower, while macrophage numbers were higher in the IR group receiving lidocaine compared to the LRS horses (P<0.05). We conclude that intestinal IR leads to remote lung injury characterized by recruitment of inflammatory cells and expression of inflammatory molecules in horses, and lidocaine may ameliorate lung inflammation following intestinal IR.