Evaluation of Nuclear Parameters in Relation to Regional Lymph Node Involvement in Oral Squamous Cell Carcinoma: A Cytomorphometric Study

Background: Up-regulation of ribosome biogenesis encodes the factors related to carcinogenesis. It has been shown that the nucleus diameter and number of nucleoli increase from normal mucosa to oral squamous cell carcinoma (OSCC). The relationship between nuclear parameters and lymph node involvement in OSCC has not been established, yet. The aim of this study was to evaluate the nuclear parameters comprising nucleoli count and nucleus: nucleoli ratio in relation to regional lymph node involvement in OSCC. Methods: Thirty-four formalin-fixed, paraffin embedded sections from different histopathologic grades of OSCC were stained with methyl green-pyronin. Mean number of nucleoli and nucleus: nucleoli ratio were calculated in 100 tumor cells from 10 random selected fields and compared based on lymph node involvement. Results: Nucleoli count in cases with metastasis to regional lymph nodes was not significantly different from that in cases without regional lymph nodes involvement (P = 0.29). The difference of nucleus: nucleoli ratio in cases with and without lymph nodes involvement were not significant (P=0.52). No significant correlation was found between the nucleoli count and lymph node involvement (r=0.08, P=0.78). The correlation between nucleus: nucleoli ratio and lymph node involvement was significant (r=0.58, P=0.02). Conclusion: The nucleus: nucleoli ratio of tumoral cells in OSCC was correlated to lymph node involvement. Based on the results, nucleus: nucleoli ratio can potentially be a useful tool to determine the lymph node involvement in OSCC

AGS cell line xenograft tumor as a suitable gastric adenocarcinoma model: growth kinetic characterization and immunohistochemistry analysis

Objective(s): Gastric cancer is the third leading cause of cancer-related death worldwide. The overall survival rate of patients is poor because gastric cancers are usually diagnosed at the late stages. Therefore, further research is needed and appropriate research tools are required to develop novel therapeutic approaches.Materials and Methods: Eight female athymic nude mice with a C57BL/6 background were used in this study. AGS cells were inoculated into the flank. The tumor volumes were calculated and growth curves were drawn. When the volume of the tumors reached 1000 mm3, the animals were humanely euthanized with CO2 gas. After harvesting, tumors were analyzed with Hematoxylin and Eosin (H&E) and immunohistochemistry (IHC). A pathologist confirmed tumor entity through H&E staining. Tumors were evaluated for expression of HER-2, P53, Ki-67, CD34, cytokeratin 8 (CK8), vimentin, estrogen receptor (ER), and progesterone receptor (PR) utilizing immunohistochemistry.Results: The tumor take rate was 62.5%, mean doubling time was 40.984 d, and the latency period was 30.62 days. The H&E staining results showed highly malignant hyperchromatin epithelial cells. IHC assessment showed the mutation status of P53 gene. The expression score of the CK8 protein in the tumor cells was +3. Vimentin protein was not expressed and changes in mesenchymal phenotype were not observed. Ki-67 IHC indicated that the proliferation rate was >43% and angiogenesis was defined as high MVD.Conclusion: The respective AGS xenograft model provides an opportunity to understand the pattern of tumor growth as well as to evaluate new gastric cancer therapies in in vivo studies

BRAF-V600E Protein Expression in Canine Malignant Cutaneous Melanoma, in Accordance with the Introduction of Biomarkers in Comparative Oncology Studies

Background: Melanoma is the cause of death of 1.3% of all cancer patients in humans. The key role of BRAF protein in the progression of human melanoma has been confirmed and its prognostic significance has been revealed. Because canine cancer resembles human cancer in biological behavior and molecular abnormalities, BRAF protein may be expressed in canine melanoma, the same as human melanoma. despite the investigation of BRAF mutation in canine melanoma, the status of BRAF at the protein level in canine skin melanoma has not yet been examined.   Methods: Thirty-two formalin-fixed, paraffin-embedded tissue samples of canine malignant cutaneous melanoma were randomly selected. After cutting into 3-μm-thick sections, the samples were evaluated for BRAF protein expression by immunohistochemistry and using the anti-BRAF V600E (VE1) mouse monoclonal antibody.   Results: The BRAF status was assessed using the Allred scoring system. Among the 32 samples examined, 21 samples were negative and 11 cases showed high BRAF protein expression.   Conclusion:  The detection of positive BRAF expression in 34.3% of canine cutaneous melanoma samples could be a step forward to improve treatment options, use the dog as an animal model in human melanoma clinical trials, and possibly identify a new prognostic biomarker in canine melanoma

The protective effect of nano-curcumin in experimental model of acute pancreatitis: The involvement of TLR4/NF-kB pathway

Objective(s): The objective of the present study is to explore whether Nanocurcumin improves pancreatic inflammation through the inhibition of the TLR4/NFkB signaling pathway in cerulein-induced acute pancreatitis. Methods: Acute pancreatitis was induced by five intraperitoneal (i.p.) injection of cerulein (50 μg/kg) with 1h intervals. Vehicle and nanocurcumin (100mg/kg/day) were given to the animals by oral gavage six days before the induction of pancreatitis. The last dose was administered 1 hour before pancreatitis induction. The serum level of amylase and lipase and the tissue level of MPO enzymes were assessed by biochemical analysis. Microscopic lesions were examined. In addition, the expression level of TLR4, NF-kB p65 and TNF-α proteins were measured by western blotting analysis. Results: Nanocurcumin reduced the microscopic lesions. In addition, the drug decreased the level of amylase, lipase and MPO enzymes. Furthermore, nanocurcumin inhibited the cerulein-induced expression of TLR4, NF-kB p65 and TNF-α proteins.   Conclusion: It is suggested that the anti-inflammatory effect of nanocurcumin on cerulein-induced acute pancreatitis may involve the inhibition of the TLR4/NFkB signaling pathway

Evaluation of antitumor activity of a TGF-beta receptor I inhibitor (SD-208) on human colon adenocarcinoma.

BACKGROUND: Transforming growth factor-β (TGF-β) pathway is involved in primary tumor progression and in promoting metastasis in a considerable proportion of human cancers such as colorectal cancer (CRC). Therefore, blockage of TGF-β pathway signaling via an inhibitor could be a valuable tool in CRC treatment. METHODS: To evaluate the efficacy of systemic targeting of the TGF-β pathway for therapeutic effects on CRC, we investigated the effects of a TGβRI (TGF-β receptor 1) or TβRI kinase inhibitor, SD-208, on SW-48, colon adenocarcinoma cells. In this work, in vitro cell proliferation was studied by methyl thiazolyl tetrazolium (MTT) and bromo-2'-deoxyuridine (BrdU) assays. Also, the histopathological and immunohistochemical evaluations were conducted by hematoxylin and eosin, and Ki-67 and CD34 markers were stained, respectively. RESULTS: Our results showed no significant reduction in cell proliferation and vessel formation (170 ± 70 and 165 ± 70, P > 0.05) in treated SW-48 cells with SD-208 compared to controls. CONCLUSION: Our data suggested that SD-208 could not significantly reduce tumor growth and angiogenesis in human colorectal cancer model at least using SW-48 cells

Acute Transplantation of Human Olfactory Mucosa-Derived Olfactory Ensheathing Cells Fails to Improve Locomotor Recovery in Rats

Olfactory ensheathing cells-based therapy for spinal cord injury (SCI) repair has been a possible treatment for clinical study because of their safety in autologous transplantation and potential regenerative capability. However, there are contradictory reports on the results after transplantation in animal models. The purpose of this research was to investigate the effect of acute transplantation of human mucosa-derived olfactory ensheathing cells (OECs) on the repair of the spinal cord. Human olfactory ensheathing cells were isolated from the human mucosa and cultured under supplemented neuronal cell culture medium. They were characterized by immunocytochemistry for olfactory ensheathing cell markers. We induced spinal cord injury at T8-T9 of rats by aneurysm clips and simultaneously injected two million OECs into subarachnoid space of spinal cord. Sensory and motor behaviors were recorded by tail-flick reflex (TFR) and BBB scores, respectively every week for seven weeks after injury. Morphology and S100-beta antigen expression in olfactory ensheathing cells of the human olfactory mucosa was confirmed by immunostaining. OECs transplantation did not recover inflammation, neuronal vacuolation, hemorrhage, and cyst formation. These findings suggest that OECs transplantation in this experimental setting did not lead to tissue regeneration to enhance locomotion. These results broaden current knowledge and are additions to the science and literature

Comparison of apoptotic bodies' count & mitotic index in oral squamous cell carcinoma with regional lymph node involvement

Background & objectives: In early stages of oral cancers, 20-40 per cent of cases have occult metastasis in cervical lymph nodes. Biologic imbalance between cellular proliferation and death culminates in metastasis. The importance of cell cycle dysregulation in relation to lymph node involvement in oral squamous cell carcinoma (OSCC) has not been established yet. The aim was to determine the association between apoptotic bodies count and mitotic index in relation to regional lymph node involvement in OSCC. Methods: Thirty two methyl green-pyronin stained slides from paraffin-embedded sections of OSCC were evaluated for apoptotic bodies count and mitotic index in relation to regional lymph node involvement using light microscopy. Number of apoptotic bodies and mitotic figures were counted in 10 randomly selected hot spot areas (×400). Average count of apoptotic bodies and mitotic figures were determined and compared with regard to the presence/absence of lymph node involvement. Results: The count of apoptotic bodies in cases without metastasis to the regional lymph node was significantly higher than in cases with regional lymph node involvement. The mitotic index was not significantly different between groups in terms of regional lymph node involvement (P=0.24). No significant correlation was found between the apoptotic bodies count (r=−0.094, P=0.72) and mitotic index (r=−0.08, P=0.75) to the number of regional lymph nodes involved. Interpretation & conclusions: Based on the results, it is suggested that apoptotic cell count can be a good parameter for showing the possibility of regional lymph node involvement in people with OSCC who do not have clinical symptoms of lymph node involvement

Development of New Therapeutic Strategies in Gynecological Cancers in Iran by Utilizing Xenograft Model of Ovarian Adenocarcinoma

Objective: To evaluate the potentiality of OVCAR–3 cell line of ovarian adenocarcinoma as a xenograft model for ovarian adenocarcinoma in nude mice. Materials and methods: The cell line isolated from advanced human ovarian adenocarcinoma, were inoculated to eight nude mice and two months later. Established tumors were transferred to pathology laboratory to be prepared by H&E staining and immunohistochemical staining with CA125 antibody. Results: Study of H&E slides showed advanced adenocarcinoma. The CA125 Tumor marker was also positive in tumoral tissue. Conclusion: Established tumors showed excellently the characteristics of ovarian adenocarcinoma. This model can be used to evaluate new treatment strategies

Feasibility assessment of in vitro chemoresponse assay on stereotactic biopsies of glioblastoma multiforms: a step towards personalized medicine

Objective(s):P In vitro chemosensitivity and resistance assays (CSRAs) are a promising tool for personalized treatment of glioblastoma multiform (GBM). These assays require a minimum of 1 to 2 g of tumor specimen for testing, but this amount is not always accessible. We aimed to assess the feasibility and validity of utilizing stereotactic biopsies of GBM in CSRAs. Materials and Methods: Single cell suspension was prepared from 1 g weight explants of the established xenograft tumor of GBM. Also, primary culture was carried out on 35 mg weight specimens, as a surrogate for stereotactic biopsies. Then, chemoresponse profile of cells obtained by direct cell disaggregation and primary culture was determined using temozolomide and carmustine by clonogenic assay[AGA1] . Results: There was no statistically significant difference in the cytotoxicity of temozolomide and carmustine between cells obtained from both methods. Conclusion: This work supports the feasibility of using stereotactic biopsies of GBM in CSRAs