Histology and histopathology Vol.23,nº12 (2008)
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- PublicationOpen AccessDecreased expression of antioxidant enzymes in the conjunctival epithelium of dry eye (Sjogren s syndrome) and its possible contribution to the development of ocular surface oxidative injuries(Murcia : F. Hernández, 2008) Cejkova, J.; Ardan, T.; Simonová, Z.; Cejka, C.; Malec, J.; Dotrelová, D.; Brunová, B.Previous studies have described elevated lipid peroxidase, myeloperoxidase and xanthine oxidoreductase/xanthine oxidase levels on the ocular surface of patients suffering from autoimmune dry eye (Sjögren’s syndrome, SS). Reactive oxygen species generated by various enzymatic systems may be dangerous to the eye if they are not sufficiently cleaved by antioxidants. Because antioxidants have not been investigated in dry eye, the aim of this study was to examine the expression of antioxidant enzymes that cleave reactive oxygen species and play a key role in antioxidant protection. Conjunctival epithelial cells of dry eye (SS) patients were obtained by the method of impression cytology using Millicell membranes. Normal eyes served as controls. In the conjunctival epithelium superoxide dismutase, catalase and glutathione peroxidase were examined immunohistochemically. The enzyme expression levels were determined by image analysis and statistical evaluation. In contrast to normal eyes, where antioxidant enzymes were highly expressed in the conjunctival epithelium, in dry eye their expression was much less pronounced in correlation with the increasing severity of dry eye symptoms. Our study suggests that the decreased expression of antioxidant enzymes in dry eye disease (SS) contributes to the development of anterior eye surface oxidative injuries.
- PublicationOpen AccessExpression of ADAMTS-5 in deformed human temporomandibular joint discs(Murcia : F. Hernández, 2008) Matsumoto, Takashi; Tojyo, Itaru; Kiga, Norifumi; Hiraishi, Yukihiro; Fujita, ShigeyukiObjective: To study the expression of a disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS-5) in tissue samples of deformed human temporomandibular joint (TMJ) discs and cells obtained from the discs. Materials and methods: Eleven adult human TMJ discs (nine diseased discs and two normal discs) were used in this study. The nine diseased discs were obtained from nine patients with internal derangement (ID) and osteoarthritis (OA) in the TMJ. These patients all had anteriorly displaced discs and deformed mandibular condyles, making conservative therapy impossible. The tissues were immunohistochemically stained using ADAMTS-5 antibodies. In addition, an articular disc cell line from one case was established by collagenase treatment. The subcultured cells under both normal and hypoxic conditions (O2: 2%) were incubated for 3, 6, 12 and 24 h after addition of interleukin-1ß (IL-1ß) (1 ng/mL). Subsequently, the expression of ADAMTS-5 was examined using reverse transcription-polymerase chain reaction (RT-PCR). Results: The control group showed negative reactions on immunohistochemical staining. The discs extracted from cases with ID and OA presented positive reactions for ADAMTS-5. The expression of ADAMTS-5 mRNA increased under both normoxia and hypoxia with the addition of IL-1ß, and the peak was observed after 3 h. Conclusion: These results suggest that ADAMTS-5 is related to deformation and destruction of human TMJ discs affected by ID and OA.
- PublicationOpen AccessMorphological effects of oestradiol-17ß‚ and selective oestrogen receptor a and ß agonists on luteinising hormone-secreting(Murcia : F. Hernández, 2008) Garrido-Gracia, José C.; Gordon, Ana; Aguilar, Rafaela; Monterde, J. G.; Blanco, A.; Martín de las Mulas, J.; Sánchez-Criado, J.E.Summary. To investigate the role played by the different rat gonadotroph oestrogen receptor (ER) pools in the effects of oestradiol-17ß (E2) on gonadectomy cells, two-week ovariectomised (OVX) rats were used. The basic experimental group of rats was injected with 3 mg of the selective ER modulator tamoxifen (TX) on days 15-20 after OVX. Groups of TX-treated OVX rats were additionally injected on days 18-20 after OVX with 10 µg oestradiol benzoate (EB), 1 mg of the selective ERα agonist propylpyrazole triol (PPT), or 1 mg of the selective ERß diarylpropionitrile (DPN). Negative and positive control groups were OVX rats injected over six days after OVX with 0.2 ml oil and EB, respectively. On day 21 after OVX, anterior pituitary glands were dissected out and divided into halves. One hemipituitary was processed for light microscopy and immunocytochemistry for ßLH subunit and progesterone receptor (PR), and the other hemipituitary for ultrastructural evaluation. Results showed that: gonadotrophs were the only pituitary cell type expressing PR; treatment with TX alone shrunk gonadectomy cells and induced both reorganization of membrane-enclosed intracellular organelles and PR expression, and treatment with DPN or EB, but not PPT, reduced the agonistic morphological effects of TX. Considering that TX activates nuclear ERα, the results indicate that activation of nuclear ERα is determinant for the reversal effects of E2 on gonadotrope morphology and PR expression, and the simultaneous activation of ERß modulates the action of ERα in an inhibitory fashion.
- PublicationOpen AccessLeptin and peroxisome proliferator-activated receptors, impact on normal and disturbed first trimester human pregnancy(Murcia : F. Hernández, 2008) Toth, Bettina; Bastug, Mehmet; Scholz, Christoph; Arck, Petra; Schulze, Sandra; Kunze, Sussane; Friese, Klaus; Jeschke, UdoRecent in vitro and in vivo studies emphasize the impact of leptin, peroxisome proliferator-activated receptors (PPAR) and PPAR coactivators (retinoic X receptor a (RXR), amplified in breast cancer-3 gene (AIB3)) on placental and fetal development. Therefore, the frequency and distribution pattern of PPAR, RXR, AIB3 and leptin expression in normal human first trimester pregnancy, miscarriage and hydatidiform mole was investigated by immunohistochemistry and double immunofluorescence staining. Enhanced expression of PPARß/d, RXR and AIB3 was identified in miscarried placentas. With regard to hydatidiform mole, increased expression of PPARg and PPARß/d was observed, whereas RXR was significantly down-regulated. Leptin expression was lowest in miscarriage and highest in mole pregnancies. In contrast to trophoblast tissue, expression of leptin in glandular epithelial cells of the decidua was increased in miscarriage. PPAR and leptin expressing cells at the feto-maternal interface were identified as extravillous trophoblast (EVT) by double immunofluorescence and CK7 staining. In summary, significantly reduced leptin expression was accompanied by enhanced PPARß/d, RXR and AIB3 expression in miscarried placentas. However, in mole pregnancy, up-regulation of leptin and increased expression of PPAR was detected. RXR, on the other hand, was down-regulated in mole decidua. So far, the study results implicate strong regulatory interaction of PPARs, their coactivators and leptin in human placentas. PPAR and leptin are potential targets for new treatment strategies concerning pregnancy disorders, such as miscarriage. The increasing knowledge about the role of PPARs and leptin in normal and disturbed pregnancy may help to improve pregnancy outcome.
- PublicationOpen AccessTemporal and quantitative analysis of expression of metalloproteinases MMPs and their endogenous inhibitors in atherosclerotic lesions(Murcia : F. Hernández, 2008) Yu, Ying; Koike, Tomonari; Kitajima, Shuji; Liu, Enqi; Morimoto, Masatoshi; Shiomi, Masashi; Hatakeyama, Kinta; Asada, Yujiro; Wang, Ke-Yong; Sasaguri, Yasuyuki; Watanabe, Teruo; Fan, JianglinMatrix metalloproteinases (MMPs) play an important role in the pathogenesis of vascular diseases, such as atherosclerosis, plaque rupture and aneurysms. Although several MMPs have been demonstrated in the lesions of atherosclerosis, their expression profiles during the initiation and progression of lesions have not been fully determined. We hypothesized that the expression of various MMPs, along with their endogenous inhibitors, may be differentially regulated dependent upon the lesion progression. Therefore, we made a temporal and quantitative analysis of the mRNA and protein expression of MMPs and tissue inhibitors of metalloproteinases expressed in the different stages of atherosclerotic lesions of rabbits and humans. We found that MMP-1, MMP-12 and MMP-13 expression was nearly absent in the normal arterial wall, but was remarkably increased with lesion progression. Furthermore, the expression of these MMPs in the lesions was closely associated with intimal macrophages and monocyte chemoattractant protein-1 expression, suggesting that the intimal macrophages are the major source of production of these MMPs. MMP-3 and MT1- MMP were also significantly upregulated in the earlystage lesions and fatty streaks compared to the normal aortas of rabbits. Our results indicate that MMP-1, -12, and -13 derived from intimal macrophages may play a pivotal role in both lesion initiation and progression, and therefore are potential therapeutic targets for the treatment of plaque rupture and aneurysm formation.
- PublicationOpen AccessNutritional cellular biomarkers in early life stages of fish(Murcia : F. Hernández, 2008) Gisbert, E.; Ortiz-Delgado, J.B.; Sarasquete, C.Histological (tissular and cellular) indices have a tradition of determining the nutritional condition of fish both in the laboratory and in the wild. The assessment of condition by means of microscopical methods is probably the mos accurate indicator of nutritional status during the early life stages of fish. This success is partly attributable large amount of information that can be derived from their study and because they are thought to be the only true starvation indices. The technique usually consists of the examination of cells and organs and the establishment of a grading system based on the presence/absence of standardised biomarkers. Each organ is examined, and the cellular aspect or tissular cohesion is evaluated qualitatively and even quantitatively in order to obtain a measure of the general condition of a larva. The literature indicates that there are certain tissular and cellular responses to food availability and quality, particularly in the digestive and muscular tissues, which are common to most teleost fish larvae. These responses, which are independent of water temperature, can be used for assessing fish larvae nutritional condition. In this regard, the microscopical organization of the liver hepatocytes, the intestinal mucosa, the exocrine pancreas and the muscular fibers, which are generally used as target tissues and organs to assess the nutritional condition of fish larvae, is deeply reviewed. The advantages and disadvantages of the use of different cellular biomarkers of effect are discussed considering different conditions.
- PublicationOpen AccessExtensive alteration in the expression profiles of TGFB pathway signaling components and TP53 is observed(Murcia : F. Hernández, 2008) Seok-Hyung Kim; Seung-Hyun Lee; Yoon-La Choi; Li-Hui Wang; Cheol Keun Park; Young Kee ShinSummary. Aims: The expression patterns of TGFB signaling proteins, such as TGFB1/2, TGFBR1(ALK5), TGFBR2, SMAD1/2/3, SMAD2/3, SMAD4, SMAD7, and of downstream targets of TGFB signaling, CDKN1A (p21CIP1), CDKN1B (p27KIP1), MYC, CDC25A, TP53, and RELA (p65NF-kB) were investigated in gastric carcinomas and other gastric lesions. Methods and results: A total of 112 gastric carcinomas, 37 dysplasias, 54 intestinal metaplasias, 29 chronic atrophic gastritis and 54 normal gastric epithelium were analyzed by tissue microarray-based immunohistochemical analysis. Extensive changes in expression profiles of these proteins were observed. Three types of expression patterns were observed along the normal epithelium-atrophic gastritis-dysplasiacarcinoma sequence. (1) Expression of TGFB1/2, TGFBR1, MYC, and TP53 continually increased along this sequence. (2) Expression of SMAD4, CDKN1A, SMAD1/2/3, SMAD2/3, and CDKN1B was enhanced in dysplasia but decreased in carcinoma. (3) Expression of TGFBR2, SMAD7, RELA, and CDC25A was enhanced in dysplasia and the enhanced level was maintained in carcinoma. In addition, we also evaluated the clinical significance of the expression of TGFB signaling proteins in gastric carcinoma. TGFB and MYC were positively correlated with advanced stages, whereas SMAD1/2/3 and SMAD4 were strongly associated with earlier stages. Conclusions: The extensive change in expression of TGFB signaling components is implicated during tumorigenesis of gastric neoplasias.
- PublicationOpen AccessReduced expression of sarcospan in muscles of Fukuyama congenital muscular dystrophy(Murcia : F. Hernández, 2008) Yoshihiro Wakayama; Masahiko Inoue; Hiroko Kojima; Sumimasa Yamashita; Seiji Shibuya; Takahiro Jimi; Hajime Hara; Yoko Matsuzaki; Hiroaki Oniki; Motoi Kanagawa; Kazuhiro Kobayashi; Tatsushi TodaSummary. Expression profiles of sarcospan in muscles with muscular dystrophies are scarcely reported. To examine this, we studied five Fukuyama congenital muscular dystrophy (FCMD) muscles, five Duchenne muscular dystrophy (DMD) muscles, five disease control and five normal control muscles. Immunoblot showed reactions of sarcospan markedly decreased in FCMD and DMD muscle extracts. Immunohistochemistry of FCMD muscles showed that most large diameter myofibers expressed sarcospan discontinuously at their surface membranes. Immature small diameter FCMD myofibers usually did not express sarcospan. Immunoreactivity of sarcospan in DMD muscles was similarly reduced. With regard to dystroglycans and sarcoglycans, immunohistochemistry of FCMD muscles showed selective deficiency of glycosylated α- dystroglycan, together with reduced expression of ß- dystroglycan and α-, ß-, γ-, δ-sarcoglycans. Although the expression of glycosylated α-dystroglycan was lost, scattered FCMD myofibers showed positive immunoreaction with an antibody against the core protein of α-dystroglycan. The group mean ratios of sarcospan mRNA copy number versus GAPDH mRNA copy number by real-time RT-PCR showed that the ratios between FCMD and normal control groups were not significantly different (P>0.1 by the two-tailed t test). This study implied either O-linked glycosylation defects of α-dystroglycan in the Golgi apparatus of FCMD muscles may lead to decreased expression of sarcoglycan and sarcospan molecules, or selective deficiency of glycosylated α-dystroglycan due to impaired glycosylation in FCMD muscles may affect the molecular integrity of the basal lamina of myofibers. This, in turn, leads to decreased expression of sarcoglycans, and finally of sarcospan at the FCMD myofiber surfaces.
- PublicationOpen AccessRAD50-MRE11-NBS1 proteins in relation to tumour development and prognosis in patients with microsatellite stable colorectal cancer(Murcia : F. Hernández, 2008) Gao, Jingfang; Zhang, Hong; Arbman, Gunnar; Sun, Xiao-FengRAD50/MRE11/NBS1 complex is essential for DNA double-strand break repair and for maintaining genomic integrity. In this study, we immunohistochemically examined MRE11, NBS1 and RAD50 expression in primary CRCs (n=208), the corresponding distant (n=41) and adjacent normal mucosa (n=130), and lymph node metastases (n=26), and investigated their clinicopathological significance in colorectal cancers (CRCs). We found that the intensity and percentage of MRE11 and NBS1 in primary CRCs were positively correlated with each other and with RAD50 (P<0.0001). Strong expression of MRE11, NBS1 or combined RAD50/MRE11/NBS1 was related to MSS, positive hMLH1 expression, earlier tumour stage (TNM stage I and II) and favourable survival (P<0.05). A high percentage of MRE11 expression was associated with less local recurrence and high apoptotic activity (P<0.05). In MSS CRCs, the expression of MRE11 and NBS1 was stronger than that in normal mucosa (P<0.05), and strong expression of NBS1 in primary tumour was related to favourable survival of patients in TNM stage I and II (univariate analysis: P=0.03; multivariate analysis: P=0.07). In MSI CRCs, neither MRE11 nor NBS1 expression showed differences among normal mucosa, primary tumour and metastasis, or among clinicopathological variables. In conclusion, RAD50/MRE11/NBS1 proteins interacted with each other, which had different clinicopathological significance in MSS and MSI CRCs, and further, each component of the complex might have additional roles. NBS1 might be a prognostic factor for patients with MSS tumour in TNM stage I and II.
- PublicationOpen AccessPrimordial germ cell specification, the importance of being blimped(Murcia : F. Hernández, 2008) Chuva de Sousa Lopes, Susana M.; Roelen, Bernard A.J.In mouse embryos, the expression of Blimp1 has recently revealed a population of allocated primordial germ cell precursors 24 hours earlier than previously thought. Those ‘blimped’ precursors have been shown to give rise, by mitotic division, to germ cells only and no other cell lineages. Here, we try to understand the events that lead to Blimp1 expression in the primordial germ cell precursors and speculate on what can be the role of Blimp1 during primordial germ cell specification and gastrulation in the mouse. Finally, we discuss the possible involvement of Blimp1 in the two know modes of germ line segregation (epigenesis and preformation).
- PublicationOpen AccessMucinous tubular and spindle cell carcinoma with Fuhrman nuclear grade 3, A histological, immunohistochemical, ultrastructural and FISH study(Murcia : F. Hernández, 2008) Kuroda, Naoto; Hes, O.; Michal, Michal; Nemcova, J.; Gal, V.; Yamaguchi, T.; Kawada, T.; Imamura, Y.; Hayashi, Yoshihiro; Lee, Gang-HongMucinous tubular and spindle cell carcinoma (MTSCC) of the kidney generally shows low nuclear grade. MTSCC with high nuclear grade is relatively rare. In this article, we report two cases of MTSCC with Fuhrman grade 3. One case occurred in a 57-year-old Japanese female and the second case in a 49-year-old Caucasian female. Histologically, the tumors were composed of neoplastic cells with cuboidal or columnar and spindle morphology, and Fuhrman nuclear grade 3. The myxoid stroma was also observed. This stroma was positive for Alcian blue stain. Immunohistochemically, neoplastic cells of both cases were positive for AMACR, but negative for CD10 and RCC Ma. Ultrastructurally, tumorous cells of one case contained numerous mitochondria. In FISH analysis, many neoplastic cells of both cases demonstrated monosomy of chromosomes 15 and 22 and disomy of chromosomes 7 and 17. One of the two patients died of respiratory failure due to pleuritis carcinomatosa 48 months postoperatively. Finally, the pathologist should recognize that high grade MTSCC exists despite its rare frequency. FISH analysis may be helpful in establishing the diagnosis of this entity. Furthermore, we present the first report of a patient with MTSCC dying of distant metastasis.
- PublicationOpen AccessThe MUC gene family, Their role in the diagnosis and prognosis of gastric cancer(Murcia : F. Hernández, 2008) Senapati, Shantibhusan; Sharma, Poonam; Bafna, Sangeeta; Roy, Hemant K.; Batra, Surinder K.Early diagnosis of gastric cancer and its differential diagnosis from other non-malignant gastric diseases like gastritis is still a major clinical problem. Most patients are asymptomatic in the early stages of gastric cancer, and there is no reliable marker available for the early and specific diagnosis of gastric cancer. Many attempts have been made to define the biological profile of gastric cancer to improve the chances of its early diagnosis, prognosis and treatment. Several studies have shown the aberrant expression profile of mucins in different malignancies, suggesting that mucins have a great potential to be used as a diagnostic and prognostic marker in gastric cancer. In this review, we have briefly described the different types of gastric adenocarcinomas and the progression of gastric cancer. Furthermore, the role of mucins and their related carbohydrate epitope is discussed in the normal stomach and in the diagnosis and prognosis of gastric adenocarcinomas.