Association between aryl hydrocarbon receptor and 4-hydroxynonenal in oxidative stress-mediated chronic rhinosinusitis with nasal polyps

Background: Сhronic rhinosinusitis with nasal polyps (CRSwNPs) is a distinct entity within the chronic rhinosinusitis group of diseases, which are chronic upper airway diseases with several pheno- and endotypes. Oxidative stress plays an important role in the pathogenesis of CRSwNPs. Aim: The aim was to assess the association between the expression of the aryl hydrocarbon receptor (AhR) and 4-hydroxynonenal (4-HNE) in patients with CRSwNPs. Methods: The study included 26 patients who underwent endoscopic sinus surgery – 14 patients with CRSwNPs, and 12 controls with healthy sinus mucosa. The expression of AhR and 4-HNE was assessed in tissue samples using immunohistochemistry. The level of 4-HNE in serum samples was measured using the ELISA assay. The total oxidative capacity (TOC) was assessed by measuring the peroxidase activity. Results: Higher levels of 4-HNE expression were observed in tissues (3, range 1-3 vs. 0, range 0-0, p<0.001) and serum (27.7±11.5 vs. 9.8±7.7 pmol/mg, p<0.001) samples of CRSwNPs patients, as compared to healthy controls. A higher expression of AhR was found in inflammatory cells (plasma cells, lymphocytes, eosinopholes) of CRSwNPs patients, compared to controls (3, range 1-3 vs. 2, range 1-2, p=0.001). There were no differences in TOC across groups (0.0285±0.0207 vs. 0.02978±0.0197 µM H2O2 eq., p=0.848). Patients with bronchial asthma (57%) had abundant eosinophiles in tissue samples. Patients with recalcitrant CRSwNPs had higher 4-HNE serum levels, compared to non-recalcitrant cases (27.3 vs. 24.2 pmol/mg, p=0.339). Conclusion: Patients suffering from CRSwNPs have oxidative stress mediated overexpression of AhR, which is linked to a chronic inflammatory response in the paranasal sinus tissues

Merkel Cell Carcinoma of the Head and Neck and Associated Second Primary Cancers: Report of Three Cases

Merkel cell carcinoma (MCC) is a rare and aggressive neuroendocrine skin carcinoma¹. The purpose of this study is to describe clinical and pathological characteristics, diagnostic procedure and treatment outcomes of the patients with MCC of the head and neck treated in Otorhinolaryngology, Head and Neck Surgery Department of the University Hospital Center Zagreb between the years 2007 and 2011. Three patients with MCC of the head and neck were treated during this period. First patient was diagnosed with MCC of the left retroauricular region with metastases in the lymph nodes of the left side of the neck, pT2N2M0 Stage IIIB. Second patient was diagnosed with anaplastic carcinoma of the lower lip, pT1N1bM0 Stage IIIB and third patient was diagnosed with MCC of the face which was previously treated as benign lesion, cT1N1bM0 Stage IIIB. Two of the patients had second primary tumor of different histology. All of the patients were treated with wide surgical excision of the tumor and neck dissection combined with adjuvant radiotherapy. Treatment outcome was poor and reason for this was late detection of disease. Menagment of the MCC patients requires multidisciplinary approach with high clinical suspicion of the treating specialist and pathologist due to immunohistochemical techniques required for diagnosis. Detection of the MCC in earlier stages is necessary for the better survival rate

C-myc Expression in the Microvessels of Medulloblastoma

The increased expression of c-myc is related to neoplastic transformation and angiogenesis. Therefore, the assessment of expression of c-myc in endothelial cells and neovascularization could help to determine the biological behavior of the tumor. We analyzed neovascularization and c-myc expression in 36 medulloblastoma specimens. The results were shown by determining immunohistochemical staining index (ISI), the sum of staining intensity (SI) and the percentage of positive cells (PPC) in the blood vessels endothelium of the tumor. We also performed the microvessel count (MVC) in 10 high-power fields (400X) with the most prominent vascularization and expressed it as microvessel density per mm2 (MVD). C-myc immunostaining intensity index in blood vessel endothelium is grouped into four groups, 0 – no reaction, I – weak reaction (ISI = 1 or 2), II – moderate reaction (ISI = 3 or 4), III – strong reaction (ISI = 5 or 6). Statistically significant differences (p=0.0214) have been found between groups 0 and 1 compared to groups 2 and 3. A higher percentage of positive cells has been found in male patients than in female ones (p=0.0483). C-myc PPC 0 or 1 has on the average smaller density of blood vessels per mm2 than c-myc PPC 2 or 3, but the difference is not statistically significant. C-myc ISI 0 or 1 has, on the average, smaller density of blood vessels per mm2 than c-myc ISI 2 or 3, but the difference is not statistically significant.We concluded that c-myc staining intensity was associated with higher microvessels density

The Appearance of 4-Hydroxy-2-Nonenal (HNE) in Squamous Cell Carcinoma of the Oropharynx

Tumor growth is associated with oxidative stress, which causes lipid peroxidation. The most intensively studied product of lipid peroxidation is 4- hydroxy-2-nonenal (HNE), which is considered as a “second messenger of free radicals” that binds to proteins and acts as a growth-regulating signaling factor. The incidence of squamous cell carcinoma of the oropharynx is associated with smoking, alcohol and infection of human papilloma virus (HPV), with increasing incidence world-wide. The aim of this retrospective study involving 102 patients was to determine the immunohistochemical appearance of HNE-protein adducts as a potential biomarker of lipid peroxidation in squamous cell carcinoma of the oropharynx. The HNE-protein adducts were detected in almost all tumor samples and in the surrounding non-tumorous tissue, while we found that HNE is differentially distributed in squamous cell carcinomas in dependence of clinical stage and histological grading of these tumors. Namely, the level of HNE-immunopositivity was increased in comparison to the normal oropharyngeal epithelium in well- and in moderately- differentiated squamous cell carcinoma, while it was decreasing in poorly differentiated carcinomas and in advanced stages of cancer. However, more malignant and advanced cancer was associated with the increase of HNE in surrounding, normal tissue. This study confirmed the onset of lipid peroxidation, generating HNE-protein adducts that can be used as a valuable bioactive marker of carcinogenesis in squamous cell carcinoma of the oropharynx, as well as indicating involvement of HNE in pathophysiological changes of the non-malignant tissue in the vicinity of cancer

Positron emission tomography-computed tomography and 4-hydroxynonenal-histidine immunohistochemistry reveal differential onset of lipid peroxidation in primary lung cancer and in pulmonary metastasis of remote malignancies

The Aim of the study was to reveal if PET-CT analysis of primary and of secondary lung cancer could be related to the onset of lipid peroxidation in cancer and in surrounding non-malignant lung tissue

Expression and prognostic value of putative cancer stem cell markers CD117 and CD15 in choroidal and ciliary body melanoma

AIMS: The aim of the present study was to immunohistochemically investigate the expression and prognostic significance of putative cancer stem cell markers CD117 (c-kit), CD34, CD20 and CD15 in a cohort of patients with primary choroidal and ciliary body melanoma. ----- METHODS: The immunohistochemical expression of these markers was evaluated using 3,3'-diaminobenzidine tetrahydrochloride (DAB) and 3-amino-9-ethylcarbazole (AEC) chromogens on paraffin-embedded tissue samples from 40 patients who underwent enucleation in the period from 1985 through 2000. Thirty-one patients had adequate tissue specimens for the analysis. ----- RESULTS: CD117 overexpression was observed in 12 of the 31 samples (39%) when AEC chromogen was used and in 14 of 26 (54%) samples when DAB was used. CD15 positivity was seen in three out of 30 (10%) samples with AEC and in six out of 26 (23%) samples with DAB. CD20 and CD34 exhibited no positivity in the tested samples. During the average follow-up time of 8.7 years (range 0.5-22 years), 17 patients (55%) died due to metastatic disease. The Kaplan-Meier plots showed a significantly shorter overall and disease-free survival in CD117-positive patients when the AEC chromogen was used. CD15 expression was not associated with patients' survival. In multivariate analysis, patients expressing the CD117 AEC had 4.13 times higher risk of lethal outcome in comparison with CD117 AEC negative patients. ----- CONCLUSIONS: Our retrospective cohort study has for the first time demonstrated a small proportion of CD15-positive uveal melanomas. CD117 AEC overexpression was associated with a worse outcome in patients with choroidal and ciliary body melanoma. Further studies should confirm the validity of these observations and their potential for targeted treatment modalities

Diagnostic Pitfalls in Parathyroid Gland Cytology

The aim of this study is to establish possibilities of using cytology in the diagnosis of parathyroid gland adenoma. 475 patients, all suspected to have parathyroid gland disease, were examined over a three-year period (from 1 of January 2006 to 31 of December 2008) in the Clinical Department of Nuclear Medicine and Radiation Protection, University Hospital Center Zagreb, Croatia. Ultrasound guided fine needle aspiration biopsy (UG-FNAB) of suspected occurrences determined by ultrasound was done. Samples obtained by UG-FNAB were air-dried and stained using the May-Grünwald- -Giemsa (MGG) staining procedure. PTH levels were determined in all punctate and sera obtained on the day of UG- -FNAB. Samples adequate for cytological analysis were obtained from 288 patients, while 187 punctates did not contain epithelial elements. The parathyroid hormone (PTH) analysis was made for all punctates. The adenoma was diagnosed via morphological characteristics in 71 out of 288 punctates that were proven adequate for cytological analysis. Increased PTH levels were later on established in all diagnosed adenomas. All patients with cytology-based diagnosis of parathyroid gland adenoma were sent to surgery, and the cytological diagnosis was confirmed by pathohistology. In three cases, the parathyroid gland adenoma was established by pathohistology, although in these cases the cytological diagnosis was negative. The cytological diagnosis of parathyroid gland adenoma can be considered reliable in 96% of cases, provided that the echosonographic structure and localisation of the punctured node is noted, and assuming that material adequate for cytological analysis is obtained by FNAB. Possible pitfalls are oncocytic types of parathyroid adenoma, intranuclear inclusions and papillary formation of epithelial cells, and cystic degeneration of nodules. These errors can be avoided by defining the PTH level on the same punctate