The novel role of HtrA1 in gingivitis, chronic and aggressive periodontitis.

Proteolytic tissue degradation is a typical phenomenon in inflammatory periodontal diseases. HtrA1 (High temperature requirement A 1) has a serine protease activity and is able to degrade fibronectin whose fragments induce the expression and secretion of several matrix metalloproteinases (MMPs). The aim of this study was to investigate for the first time if HtrA1 has a role in gingivitis and in generalized forms of chronic and aggressive periodontitis. Expression of HtrA1 was investigated in 16 clinically healthy gingiva, 16 gingivitis, 14 generalized chronic periodontitis and 10 generalized aggressive periodontitis by immunohistochemistry and real-time PCR. Statistical comparisons were performed by the Kruskall-Wallis test. Significantly higher levels of HtrA1 mRNA and protein expression were observed in pathological respect to healthy tissues. In particular, we detected an increase of plasma cell HtrA1 immunostaining from gingivitis to chronic and aggressive periodontitis, with the higher intensity in aggressive disease. In addition, we observed the presence of HtrA1 in normal and pathological epithelium, with an increased expression, particularly in its superficial layer, associated with increasingly severe forms of periodontal disease. We can affirm that HtrA1 expression in plasma cells could be correlated with the destruction of pathological periodontal tissue, probably due to its ability to trigger the overproduction of MMPs and to increase the inflammatory mediators TNF-α and IL-1ß by inhibition of TGF-ß. Moreover, epithelial HtrA1 immunostaining suggests a participation of the molecule in the host inflammatory immune responses necessary for the control of periodontal infection.

The involvement of TGF-ß1 and CTGF in regional gingival overgrowth.

Gingival overgrowth is a multifactorial and invalidating condition. Our research is about gingival overgrowth caused by gingival plaque, its purpose being the evaluation of the presence of gingivitis and/or parodontitis in patients with gingival growth and the extent in which there is a connection between gingival overgrowth and the inflammatory process that can contribute to an exceedingly stimulation of the overgrowth. Immunohistological study was conducted on human material--gingival mucosa that came from patients with ages between 20-65 years, divided into three groups: group I--control group, group II--patients with gingivitis, group III--patients with local or general periodontitis. The intensity of immunohistochemical staining of TGF-ß1 and CTGF varies from one group to another, and also depends on the area of gingival mucosa that was observed. TGF-ß1 has a crucial role in periodontal disease fibrogenesis by intensifying the action of CTGF.

Evidence for the involvement of TGF-ß1-CTGF axis in liver fibrogenesis secondary to hepatic viral infection.

Liver fibrosis is a nonspecific response to injuries, which implies the synthesis of an abnormal extracellular matrix (ECM). TGF-ß (transforming growth factor ß) is a cytokine involved in regulation of several important processes: cell development and differentiation, apoptosis, synthesis and degradation of ECM. CTGF (connective tissue growth factor) is a cysteine rich peptide that belongs to the CCN family of proteins and plays an essential role in the formation of blood vessels, bone and connective tissue. The purpose of this study was to assess TGF-ß1 and CTGF immunohistochemical expression in different stages of liver fibrosis secondary to chronic viral hepatitis. Liver biopsies from patients diagnosed with chronic viral hepatitis B and C were embedded in paraffin and further used for histological staining and immunohistochemical reactions to detect TGF-ß1 and CTGF. Liver sections stained with trichromic Masson for collagen staining and Gömöri's silver impregnation revealed various degrees of liver fibrosis, noted in the METAVIR scale from 1 to 4. Sections with discrete degrees of fibrosis revealed the positivity only in the endothelial cells of liver sinusoids and occasionally in proinflammatory cells from the portal tracts, the number of TGF-ß1-positive cells being directly proportional to the incidence of liver injury. Positive reaction for TGF-ß1 expanded to the cytoplasm of hepatocytes located nearby fibrosis bundles while increasing the parenchymal damage. The expression of CTGF was observed in the classical areas of the hepatic lobule, such as the perisinusoidal spaces around the portal tracts or central veins, but also in the hepatocytes surrounding the fibrotic areas. Regardless of the etiological factor of liver damage, activation of liver cells causes an increased synthesis of TGF-ß1 followed by a CTGF overproduction in various polymorphic hepatic structures, in accordance with the degree of fibrosis.

Immunohistochemical aspects of endometrial glands in dysfunctional uterine hemorrhage.

The authors present a specific aspect of the modifications of the endometrium in dysfunctional uterine hemorrhages that is the behavior of the endometrial glands. These glands are studied from a immunohistochemical point of view, regarding both the normal endometrium (inclusively at the age of two years) and the endometrium in dysfunctional uterine hemorrhages. The antigens used were VGEF and PCNA. The result was a different reaction of the glandular structures to these antigens in the cases of patients with dysfunctional uterine hemorrhage.

Idiopathic gingival hypertrophy--a morphological study and a review of literature.

UNLABELLED: Gingival overgrowth occurs as a response to several drugs, in some systemic diseases, as a consequence of a genetic predisposition or could be idiopathic, the last being less studied. Independently of the etiological factor involved, histological changes are maintained and intensified by the presence of the bacterial plaque. MATERIAL AND METHODS: Fibrotic mucosa clinically diagnosed with idiopathic hypertrophy was included in paraffin and studied using usual histological stains and immunohistochemical techniques for vimentin, MMP-3 and TIMP-2. RESULTS: Evaluation of histological changes revealed an unspecific pattern like the thickening of the epithelium, with acanthosis and acantholysis and massive deposition of connective tissue in the lamina propria. Keratinocytes from the areas with acantholysis express MMP-3. In the fibrous areas we noted few fibroblasts and a discrete expression of MMP-3. Extended areas of inflammatory tissue with numerous de novo capillaries were observed among the collagen bundles. Proinflammatory and endothelial cells expressed both MMP-3 and TIMP-2. The last antibody had an intense expression deeper in the epithelium. Our results were discussed in accordance with reference data. CONCLUSIONS: We conclude that the histological changes observed were not specific for the idiopathic gingival hypertrophy and the imbalance of tissue homeostasis was due to the interaction between keratinocytes and connective cells in the milieu of specific changes induced by the inflammatory conditions.

Immunohistochemical study of the morphological changes in placental villi from fetal membranes infectious disease.

UNLABELLED: Many types of infection cause placental changes but sometimes the etiological cause may be difficult to prove. Infections may ascend from endocervical canal or they may reach placenta hematogenously through the maternal blood. Typically, placenta of "the amnionic sac infection syndrome" is an immature placenta. Complex cellular mechanisms are involved in all types of infection that often are associated with placental insufficiency. OBJECTIVES: The aim of this study was to evaluate cellular changes induced by the hypoxic conditions due to infectious disease in the placental villous structures, especially in the trophoblast layer and vascular bed. MATERIAL AND METHODS: In order to label the trophoblast layer we used anti-cytokeratin cocktail AE1/AE3. Antibodies anti-VEGF and anti-c-ErbB4 were used for the evaluation of tissue response under hypoxic conditions and its involvement in villous remodeling. RESULTS: Chorion villi from placentas with histopathological features of insufficiency due to infectious etiology show an intense immunostaining for VEGF in the trophoblast, vessel walls and some stromal cells, namely Hofbauer cells. Villous trophoblast from the infected placenta expresses c-ErbB4 receptor. CONCLUSIONS: Overexpression of VEGF and c-ErbB4 is needed for the involvement of trophoblast layer in villous remodeling processes in order to maintain placental functionality under the effects of the inflammatory cascade.

Dual targeting of IGF-1R and PDGFR inhibits proliferation in high-grade gliomas cells and induces radiosensitivity in JNK-1 expressing cells.

Increased expression and activation of receptor tyrosine kinases frequently occur in human brain tumors, mediating a variety of growth-promoting pathways and leading to radioresistance; however, little is known about their motogenic potency relative to one another. In this study, we found co-expression of Insulin like growth factor-1 receptor (IGF-1R) and platelet derived growth factor receptor (PDGFR) in two high-grade gliomas (HGG) cell lines 18 and 38. Dual targeting of IGF-1R and PDGFR increased cell death in both 18 and 38 cell lines in comparison to inhibition of either receptor alone. In addition, co-inhibition of IGF-1R and PDGFR increased radiosensitivity in 18 cells but failed to intensify the effect of radiation in 38 cells. In HGG cells, radiation-induced cell death has been connected to the activation of c-Jun-NH2-terminal kinase-1 (JNK1). We found that JNK1 was weakly expressed in 38 cells while it had an elevated expression in 18 cells. Exposure to ionizing radiation induced JNK1 activation only in 18 cells without affecting the protein activity in 38 cells. These results suggest that in 18 cell line radiation-activated JNK1 may provide an anti-proliferative signaling, parallel to receptors co-targeting. To test this hypothesis, HGG cells were treated with dominant negative JNK1 (dnJNK1) and the response to radiation was assayed in presence or absence of receptors co-inhibition. Indeed dnJNK protected 18 cells against gamma-irradiation-induced cell death. dnJNK treatment did not influence radiation response of the 38 cell line, which expressed low levels of JNK1. In conclusion we found that IGF-1R and PDGFR co-inhibition caused an increased cell death in two HGG cell line and induced the radiosensitization of the JNK1 expressing cell line.

Immunohistochemical assessment of angiogenesis in primary hepatocellular carcinoma.

BACKGROUND: Primary hepatocellular carcinoma (HCC) is characterized by the presence of angiogenesis, which is necessary for tumor growth, progression and distant metastasis. Vascular endothelial growth factor (VEGF) is clearly expressed in HCC, in variable degrees as a function of differentiation and vascularisation. AIM: To assess angiogenesis in HCC, by means of immunohistochemical analysis of the expressions of VEGF. METHOD: Immunohistochemical techniques were performed on the samples obtained by ultrasound-guided liver biopsies or intraoperative biopsies, in 32 patients with HCC. RESULTS: Positive expression of VEGF was always observed in the extracellular matrix of portal tracts of the non-neoplastic or tumor areas (extra- and intranodular areas of HCC patients). VEGF was not expressed inside the hepatocytes of extranodular non-neoplastic areas of the patients with HCC. However, we did find positive reactions for VEGF inside the tumor hepatocytes in 34.38 % of the HCC patients. The difference between VEGF expression for the patients with poor and undifferentiated HCC as compared with moderate and well-differentiated HCC was statistically significant (P < 0.05). CONCLUSION: Our study demonstrated an increased VEGF expression in tumor hepatocytes, which progressed with the dedifferentiation of HCC. VEGF expression was always present in the extracellular matrix, supporting the hypothesis of paracrine activation of VEGF at the level of tumor stroma. Consequently, increased VEGF expression might be responsible for the activation of angiogenesis in HCC.

Immunohistochemical assessment of proliferating cell nuclear antigen in primary hepatocellular carcinoma and dysplastic nodules.

A complementary way for the assessment of HCC prognosis is represented by the analysis of molecular markers. Thus, immunohistochemical assessment of proliferation can describe tumor aggressiveness, probability of local recurrence or metastasis potential, being very useful for the assessment of recurrence-free survival and survival until death. The aim of our study was to assess proliferating cell nuclear antigen activity in HCC and dysplastic nodules as compared with surrounding non-neoplasic areas. Immunohistochemical techniques were thus performed on the samples obtained by ultrasound-guided liver biopsies or intraoperative biopsies, in 32 patients with HCC, as well as in 3 patients with dysplastic nodules occurring in liver cirrhosis. Expression of PCNA within extranodular areas of the HCC patients in the absence or presence of cirrhosis, was increasing from 40% to 70%, respectively. PCNA expression further increased within intranodular areas of dysplastic nodules and HCC, to 100% and 96.88%, respectively. A progressive increase of the mean values of PCNA-LI was also observed from extranodular areas without or with cirrhosis, towards intranodular areas of dysplastic nodules and HCC (4.2%, 6.8%, 27.9%, 31.9%, respectively). Dysplastic nodules can thus be considered lesions with a high-proliferation rate, representing an early stage of hepatocarcinogenesis. This supported the current recommendations for borderline hepatocellular nodules identified by ultrasound, which indicate an aggressive treatment similar to malignant lesions. In summary, we demonstrated a progressively increasing rate of cellular proliferation, from extranodular non-neoplasic areas to intranodular areas (dysplastic nodules and HCC), as reflected by an increased expression of proliferating cell nuclear antigen labelling index.