Histology and histopathology Vol.22, nº10 (2007)
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- PublicationOpen AccessThe melanogenic system of the liver pigmented macrophages of Rana esculenta L. - Tyrosinase activity(Murcia : F. Hernández, 2007) Gallone, A.; Sagliano, A.; Guida, G.; Ito, S.; Wakamatsu, K.; Capozzi, V.; Perna, G.; Zanna, P.; Cicero, R.The enzyme system responsible for Amphibian Kupffer Cell (KC) melanogenesis has not been entirely elucidated. This research demonstrates that the KC melanosomes of Rana esculenta L. possess a tyrosine-hydroxylase (TH) activity, showing that a tyrosinase is the enzyme involved in the melanogenesis. The TH reaction depends on catalytic Dopa as a cofactor and is not affected by catalase or H2O2, showing that it is catalysed by the tyrosinase and not by the peroxidase present in the melanosomes. The TH reaction is activated by Cu2+ ions but not by other tyrosinase activators such as limited proteolysis, protein ageing, and Sodium Dodecyl Sulphate (SDS). SDS inhibited the KC TH activity even below the critical micelle concentration. All these results suggest that the KC-tyrosinase differs in structure from other known tyrosinases. Using anti-KC-tyrosinase antobodies, we observed that the sites of the tyrosinase location within the cell are the same as those described in the melanocytes. In the immunoblots, the anti-KC-tyrosinase antibodies also recognised two protein bands, at the higher molecular weight ranges, in the protein electrophoretic pattern. Moreover, the tyrosinase activity was limited to the highest molecular weight band of about 260 kDa, suggesting that the enzyme activity could depend on a molecular aggregate. The melanin produced in the liver was found to be a 5,6-dihydroxyindole-rich eumelanin similar to the Sepia melanin.
- PublicationOpen AccessExperimental change on dopaminergic neurons in striatum of Parkinson disease rats(Murcia : F. Hernández, 2007) Chen, X.Y.; Li, Jiang; Qi, W.Q.; Shen, S.H.Background: Parkinson’s disease (PD) is a common neurodegenerative disorder characterized by selective and progressive demise of dopaminecontaining neurons in the midbrain. In this study, we observe the expression of c-Jun in the striatum of rats with 6-hydroxydopamine (6-OHDA)-lesions after apomorphine (APO) intraperitoneal injection (ip) in substantia nigra compacta (SNc), and to study the mechanism of the rotations behavior. Design: The 6- OHDA was unilaterally injected into rat right SNC. The APO- induced abnormal rotations were investigated on the 1st, 4th, 7th, 14th, 21st days after lesion, respectively. Meanwhile dopaminergic degeneration and c-Jun expression were observed with microscope. Nissl’s body staining and immunohistochemical method (ABC) were employed to study the changes of tyrosine hydroxylase (TH) and c-Jun in DA neurons. Results: We found that the number of dopaminergic neurons decreased gradually in the lesioned site and those neurons’ electron-microscopic structure was severe damaged. There were over 75% of dopaminergic neurons lost, contralateral rotations over 7 turns per minute and c-Jun expressing in the ipsilateral striatum. Conclusion: Dopaminergic neurons deletion may be linked to upregulation of c-Jun.
- PublicationOpen AccessCurrent concepts in human prion protein (Prp) misfolding, Prnp gene polymorphisms and their contribution to Creutzfeldt-Jakob Disease (CJD)(Murcia : F. Hernández, 2007) Michalczyk, K.; Ziman, M.Transmissible spongiform encephalopathies are a group of neural degenerative diseases that may be infectious, sporadic, or hereditary and are associated with an abnormally folded prion protein. Unfortunately at the current time it is not at all clear what the normal structure of the prion protein actually is or how it is toxic to cells. Extensive research on prion diseases has led to a dramatic increase in understanding of the pathogenesis of prion disorders, which will hopefully lead to the development of effective treatments. The inability to detect the disease in blood using current technology has made screening difficult. While fortunately there has been a decline in the number of clinical cases of transmissible variant CJD, evidence indicates that very long incubation periods can occur in humans so there may be a long slow, gradual epidemic. In particular, clinical cases in genotypes other than those homozygous for methionine at codon 129 of PRNP have not yet occurred, but such cases might be expected to have longer incubation periods and show differences in pathology to those seen to date. proportion of infected animals develop sub-clinical disease. Moreover, results from a large prevalence study in humans show that several cases test positive but do not develop clinical disease. It is possible therefore that further cases of secondary transmission could occur by iatrogenic spread, which could result in vCJD persisting in the UK at low levels for many years, highlighting the importance of continued vigilance.
- PublicationOpen AccessMüllerianosis(Murcia : F. Hernández, 2007) Batt, R.E; Smith, R.A.; Buck Louis, G.M.; Martin, D.C.; Chapron, C.; Koninckx, P.R.; Yeh, J.Müllerianosis may be defined as an organoid structure of embryonic origin; a choristoma composed of müllerian rests - normal endometrium, normal endosalpinx, and normal endocervix - singly or in combination, incorporated within other normal organs during organogenesis. A choristoma is a mass of histologically normal tissue that is “not normally found in the organ or structure in which it is located” (Choristoma, 2006). Müllerian choristomas are a subset of non-müllerian choristomas found throughout the body. Histologically, endometrial-müllerianosis and endometriosis are both composed of endometrial glands and stroma, but there the similarity ends. Their pathogenesis is different. Sampson faced the same difficulty with pathogenesis and nomenclature when he wrote: “The nomenclature of misplaced endometrial or müllerian lesions is a difficult one to decide upon.” “The term müllerian would be inclusive and correct, but unfortunately it suggests an embryonic origin.” Sampson then divided “misplaced endometrial or müllerian tissue” into “four or possibly five groups, according to the manner in which this tissue reached its ectopic location” (Sampson, 1925). Sampson’s classification of heterotopic or misplaced endometrial tissue is based on pathogenesis: 1) “direct or primary endometriosis” [adenomyosis]; “a similar condition occurs in the wall of the tube from its invasion by the tubal mucosa” [endosalpingiosis]; 2) “peritoneal or implantation endometriosis;” 3) “transplantation endometriosis;” 4) “metastatic endometriosis;” and 5) “developmentally misplaced endometrial tissue. (I admit the possibility of such a condition, but have never been able to appreciate it.)” (Sampson, 1925). It is precisely this condition “developmentally misplaced endometrial tissue,” [müllerianosis] that is the subject of this review.
- PublicationOpen AccessThe absence of one or both nidogens does not alter basement membrane composition in adult murine kidney(Murcia : F. Hernández, 2007) Gersdorff, N.; Otto, S.; Roediger, Matthias; Kruegel, J.; Miosge, NicolaiNidogen-1 and nidogen-2 are major components of all basement membranes and are considered to function as link molecules between laminin and collagen type IV networks. Surprisingly, the knockout of one or both nidogens does not cause defects in all tissues or in all basement membranes. In this study, we have elucidated the appearance of the major basement membrane components in adult murine kidney lacking nidogen-1, nidogen-2, or both nidogens. To this end, we localized laminin-111, perlecan, and collagen type IV in knockout mice, heterozygous (+/-) or homozygous (-/-) for the nidogen-1 gene, the nidogen-2 gene, or both nidogen genes with the help of light microscopic immunostaining. We also performed immunogold histochemistry to determine the occurrence of these molecules in the murine kidney at the ultrastructural level. The renal basement membranes of single knockout mice contained a similar distribution of laminin-111, perlecan, and collagen type IV compared to heterozygous mice. In nidogen double-knockout animals, the basement membrane underlying the tubular epithelium was sometimes altered, giving a diffuse and thickened pattern, or was totally absent. The normal or thickened basement membrane of double-knockout mice also showed a similar distribution of laminin-111, perlecan, and collagen type IV. The results indicate that the lack of nidogen-1, nidogen-2, or both nidogens, plays no crucial role in the occurrence and localization of laminin-111, collagen type IV, and perlecan in murine tubular renal basement membranes
- PublicationOpen AccessHistomorphometric study of femoral heads in hip osteoarthritis and osteoporosis(Murcia : F. Hernández, 2007) De Pedro, J.A.; Martin, A.P.; Blanco, J.F.; Salvado, M.; Perez, M.A.; Cardoso, A.; Collía, F.; Ellis, S.S.; Domínguez, J.During the period from 2000 to 2003, ninety eight samples of femoral heads were collected. In these pieces, two zones were analyzed: a high-load zone (the hard core of the head) and a low-load zone (the round ligamentum teres zone). As control group, 6 femoral heads (3 of women and 3 of men), proceeding from autopsy in peoples without pathological antecedents and youngs, were studied. After the samples had been embedded in methylmethacrylate and stained, they were subjected to an histomorphometric study. By means of histomorphometry, trabecular bone volume (TBV) and osteoid substance (OSV) was determined. Statistically significant differences were found as for peripheral osteoid volume (low-load zone) ( p=0.036) and trabecular bone volume, both peripheral and central. Both volumes decreased in osteoporotic samples and in those from women (p=0.000), in comparison with control group. Regarding the relationship between the high-load and low-load zone, significant data were obtained. The high-load zone had a greater trabecular bone volume than the low-load zone, regardless of the pathology and sex, but this increase was more pronounced in the arthrosic samples and in those from men. Additionally, this trabecular bone volume in the high-load zone decreased with increasing age of the donor (p=0.037), when the control group is compared. In sum, we observed a reduction in the formation of TBV and OSV in osteoporosis but also a decrease in the arthrosic, in samples from older subjects, in women, and in the low-load zone of the samples, when the control group is compared. These data suggest the coexistence of both pathologies, which is more pronounced in older subjects and women.
- PublicationOpen AccessOccurrence of two NOS isoforms in the developing gut of sea bass Dicentrarchus labrax (L.)(Murcia : F. Hernández, 2007) Pederzoli, A.; Conte, A.; Tagliazucchi, D.; Gambarelli, A.; Mola, L.In this work we have examined the appearance and distribution of nitric oxide synthase (NOS), with histochemical, immunohistochemical and biochemical methods, during development of the sea bass (Dicentrarchus labrax) gut. The data showed that both the calcium-calmodulin dependent neuronal isoform (nNOS) and calciumindependent inducible isoform (iNOS) are present in the larval gut of sea bass. The nNOS-immunoreactivity was present in the epithelial cells and enteric nerve cells of gut both in the 8-day-old specimens and in the 24-dayold- larvae. In the adult nNOS-immunoreactivity disappeared from epithelial cells, remaining in the wall intramural neurons and fibers. The iNOSimmunoreactivity was present in the epithelial cells of 24-day-old-larvae and was not detectable in the adult gut. Western blot analysis and determination of NOS activity also demonstrated the presence of the two NOS isoforms, nNOS and iNOS, in the gut of 24-day-old specimens. The presumably different roles played by the two isoforms of enzyme are discussed. The presence of nNOS isoform in the gut enteric neurons of the same larval stages of D. labrax in which we previously demonstrated the presence of substance P and Vasoactive Intestinal Polypeptide (VIP), may suggest that all these three components of the motility control system are already present in the larval phase. Nitric oxide (NO) may be also involved in the early immune response. The present results on the occurrence of iNOS isoform in epithelial gut cells of the same regions in which the gut-associated lymphoid tissue (GALT) will differentiate, may suggest for NO a role in early defence mechanisms, before the establishment of immune responses in GALT. Finally, the developmental and regional differences in nNOS and iNOS expession also suggest a regulatory role in development and differentiation of the sea bass gut.
- PublicationOpen AccessPeriostin: Novel diagnostic and therapeutic target for cancer(Murcia : F. Hernández, 2007) Kudo, Y.; Siriwardena, B.S.M.S.; Hatano, H.; Ogawa, I.; Takata, T.Periostin is a secreted protein that shares a structural homology to the axon guidance protein fasciclin I (FAS1) in insects and was originally named as osteoblast-specific factor-2 (Osf2). Periostin is particularly highly homologus to ßig-h3, which promotes cell adhesion and spreading of fibroblasts. It has recently been reported that Periostin was frequently overexpressed in various types of human cancers. Although the detailed function of Periostin is still unclear, Periostin-integrin interaction through FAS1 domain is thought to be involved in tumor development. In addition, Periostin stimulates metastatic growth by promoting cancer cell survival, invasion and angiogenesis. Therefore, Periostin can be a useful marker to predict the behavior of cancer. This review summarizes the recent understanding of Periostin roles in tumor development and speculates on the usefulness of Periostin as a therapeutic and diagnostic target for cancer.
- PublicationOpen AccessIncreased expression of 5-lipoxygenase is common in clear cell renal cell carcinoma(Murcia : F. Hernández, 2007) Faronato, M.; Muzzonigro, G.; Milanese, G.; Menna, C.; Bonfigli, A.R.; Catalano, A.; Procopio, A.The clinical behaviour of Clear Cell Renal Cell Carcinoma (CC-RCC) is often unpredictable. To fully understand the signaling pathways involved in CCRCC development, we examined whether the 5- Lipoxygenase (5-LO), which catalyzes the biosynthesis of proinflammatory leukotrienes, is involved in renal tumorigenesis. By analyzing 46 snap-frozen primary renal cell carcinomas and their corresponding normal renal cortex biopsies, 5-LO protein levels were found to be significantly increased in the majority of CC-RCCs (P<0.001). Quantitative 5-LO mRNA expression analysis revealed up to 3-fold increased expression in the tumor tissues. There was no association between 5-LO and gender, grade or vein invasion. In contrast, increased 5-LO protein and mRNA correlated with large tumor size (>4 cm) and age of patients (P<0.001). 5-LO was frequently overexpressed in von Hippel-Lindau protein (pVHL)–reduced tumors and in Vascular Endothelial Growth Factor (VEGF)-positive tumors, which represent two frequent alterations in CC-RCC. Cell culture experiments demonstrated that VEGF expression was strongly inducible by 5-LO metabolites in RCC cell lines. The loss of pVHL expression led to high basal 5- LO and VEGF expression, which were markedly reduced by transfection with 5-LO small interfering RNA (siRNA). These results suggest that 5-LO upregulation is an important step in renal cancer progression.
- PublicationOpen AccessBone morphogenetic proteins and their receptor signaling in prostate cancer(Murcia : F. Hernández, 2007) Ye, L.; Lewis-Russell, J.M.; Kynaston, H.G.; Jiang, W.G.Bone morphogenetic proteins (BMPs) belong to the TGF-ß superfamily and are vital bone inductive factors. BMPs also play important roles during embryonic development and the postnatal homeostasis of various organs and tissues, by controlling cellular differentiation, proliferation and apoptosis. Prostate cancer is the most common cancer in men in Western countries, with a high incidence of bone metastasis. Once bony metastasis developed, the condition is incurable, and contributes significant disease specific morbidity and mortality. However, the mechanisms underlying the development of bone metastasis remain unclear. BMPs have been implicated in the development of both primary and secondary tumors, particularly skeletal metastasis. Aberrations in BMPs signaling have also been identified in various neoplasms. Recently studies have also suggested a pivotal role in bone metastasis for Noggin, which is a BMP antagonist. In this review, we discuss the current knowledge of BMPs signaling, abnormalities which have been identified and their involvement in tumour progression, and particularly in the development of bone metastasis in prostate cancer.
- PublicationOpen AccessRole of fibrosis-related genes and pancreatic duct obstruction in rat pancreatitis models: Implications for chronic pancreatitis(Murcia : F. Hernández, 2007) Miyauchi, M.; Suda, K.; Kuwayama, C.; Abe, H.; Kakinuma, C.Human chronic pancreatitis is characterized by irreversible fibrosis, whereas pancreatic fibrosis in animal models is reversible. In this study, we compare the development of pancreatic fibrosis in the dibutyltin dichloride (DBTC) model, WBN/Kob rats and bile ductligated (BDL) rats. DBTC (8 mg/kg) was administered to LEW rats, and the pancreas was histopathologically investigated sequentially. Male and female WBN/Kob rats aged 4, 6 and 8 months were also examined. BDL rats were prepared by ligation of the bile duct at the duodenal portion and sacrificed at 3 or 7 days after ligation. Fibrosis in the DBTC model peaked after 1 week and was limited to the areas around the pancreatic ducts after 2 weeks, and was composed of both type I and type III collagen. In contrast, fibrosis in male WBN/Kob rats peaked at age 4 months, expanded into intralobular area, and was composed of type III collagen. It exhibited almost no type I collagen and a marked tendency to regress. Pancreatic fibrosis in BDL rats was somewhat difficult to induce and required increased stimulation. This suggests that fibrosis in human biliary pancreatitis may gradually form based on weak, continuous stimulation. We conclude that type I collagen may be involved in the progression of irreversible fibrosis. The imbalance between synthesis and degradation of extracellular matrix molecules or degree of stimulation over a certain period may lead to pancreatic fibrosis. Gene expressions of prolyl hydroxylase and tissue inhibitors of matrix metalloproteinase-2 were elevated.
- PublicationOpen AccessExpression of heat shock protein 70 in renal cell carcinoma and its relation to tumor progression and prognosis(Murcia : F. Hernández, 2007) Ramp, U.; Mahotka, C.; Heikaus, S.; Shibata, T.; Grimm, M.O.; Willers, R.; Gabbert, H.E.Heat shock proteins (HSPs) play an important role in the cellular response to environmental stress and exert a cytoprotective effect. Especially HSP70 is an effective inhibitor of apoptosis, suggesting a role of HSP70 in carcinogenesis and tumor progression. To explore the relevance of HSP70 in renal cell carcinomas (RCCs), we analyzed nuclear and cytoplasmic HSP70 protein expression in formalin-fixed tissue from 145 clear cell RCCs by immunohistochemistry as well as Western blot analysis. Nuclear HSP70 expression was found in all RCCs and 75% of the tumors also exhibited a cytoplasmic HSP70 staining. Importantly, RCCs showed significantly reduced cytoplasmic (p=0.001) and combined nuclear/cytoplasmic (p=0.0022) HSP70 expression when compared with their cells of origin. A significant (p=0.0176) decrease of nuclear HSP70 expression became evident from well to poorly differentiated clear cell RCCs. Quite similarly, a trend (p=0.0558) for reduced combined nuclear/cytoplasmic HSP70 expression was shown from early (pT1) to advanced (pT3) tumor stages. Nevertheless, no correlation between HSP70 expression and patients´ survival became evident. In conclusion, our investigation demonstrates a significant decrease of antiapoptotic HSP70 protein expression during carcinogenesis and during progression from well (G1) to poorly (G3) differentiated clear cell RCCs. Our results suggest that HSP70-mediated inhibition of apoptosis seems to be of minor importance for carcinogenesis and tumor progression in RCCs.
- PublicationOpen Access