Evaluation of Metalloproteinase-8 Levels in Crevicular Fluid of Patients with Healthy Implants or Periodontitis.

Evaluation of periodontal and peri-implant tissue condition is mainly based on clinical examination and imaging diagnostics. Some data imply that Metalloproteinase-8 (MMP-8) level examination in peri-implant sulcular fluid (PISF) might be useful for evaluating the condition of peri-implant tissues and monitoring a development of peri-implant inflammation, including both mucositis and peri-implantitis. Hence, in this study, we decided to evaluate the level of MMP-8 in PISF obtained from patients without clinical symptoms of mucositis or peri-implantitis and compare it with MMP-8 level in gingival crevicular fluid (GCF) obtained from patients with healthy periodontium and those with varying severity of periodontitis. A total of 189 subjects were included in the study, and GCF/PISF samples were analysed for MMP-8 level by ELISA test. We documented that MMP-8 level in PISF obtained from patients without symptoms of mucositis or peri-implantitis was significantly higher not only than in GCF of periodontally healthy patients but also, which seems to be very interesting, than in GCF of patients with varying degrees of periodontal inflammation, consistent with earlier studies. Our observation might imply that monitoring of MMP-8 level in PISF could help to diagnose mucositis/peri-implantitis in an early stage, prior to clinical manifestations, which may allow for quick start of appropriate therapy.

The c.*229C > T gene polymorphism in 3'UTR region of the topoisomerase IIß binding protein 1 gene and LOH in BRCA1/2 regions and their effect on the risk and progression of human laryngeal carcinoma.

Topoisomerase IIß binding protein 1 (TopBP1), a multiple-BRCT-domain, protein plays crucial roles in chromosome replication, DNA damage repair, apoptosis, and cell cycle checkpoint signalling. The aim of this study was to identify five SNPs at loci potentially located in the 3'UTR region of the TopBP1 gene (rs185903567, rs116645643, rs115160714, rs116195487, rs112843513), their relationship with the risk of squamous cell laryngeal cancer (SCLC), tumor invasiveness, and prognosis. Genotyping was performed in 323 genetically unrelated individuals with SCLC and 418 randomly selected healthy volunteers. Allele-specific TopBP1 mRNA and protein expressions were determined by using real-time PCR and Western blotting techniques, respectively. LOH in BRCA1/BRCA2 was determined by using microsatellite markers. Compared to homozygous common allele carriers, heterozygosity for the T variant was associated with increased risk of SCLC (adjusted odds ratio [OR] = 9.83, 95 % confidence interval [CI]: 3.12-22.16, p dominant < 0.0001). The presence of risk allele at rs115160714 TopBP1 determined a higher incidence of nodal metastases (OR = 7.98, 95 % CI: 3.94-16.00, p = 0.001) and higher tumor grade (OR = 6.48, 95 % CI: 0.86-48.01, p = 0.03). The heterozygotes displayed diffuse tumor growth with no distinct borderline (OR = 3.10, 95 % Cl: 0.92-10.62, p = 0.049) and higher depth of invasion (OR = 2.66, 95 % Cl: 0.78-9.03, p = 0.04). Relationships were also identified between TopBP1 mRNA/protein expression and overall survival (p < 0.0001). The incidence of LOH in BRCA1/BRCA2 was significantly related to higher tumor grade and TFG (p < 0.05). The results of this study suggest that rs115160714 TopBP1 may be a genetic marker of etiology and progression in laryngeal cancer.

Gene/protein expression of CAPN1/2-CAST system members is associated with ERK1/2 kinases activity as well as progression and clinical outcome in human laryngeal cancer.

Recent evidence indicates the involvement of calpains (CAPNs), a family of cysteine proteases, in cancer development and progression, as well as the insufficient response to cancer therapies. The contribution of CAPNs and regulatory calpastatin (CAST) and ERK1/2 kinases to aggressiveness, disease course, and outcome in laryngeal cancer remains elusive. This study was aimed to evaluate the CAPN1/2-CAST-ERK1/2 enzyme system mRNA/protein level and to investigate whether they can promote the dynamic of tumor growth and prognosis. The mRNA expression of marker genes was determined in 106 laryngeal cancer (SCLC) cases and 73 non-cancerous adjacent mucosa (NCLM) controls using quantitative real-time PCR. The level of corresponding proteins was analyzed by Western Blot. SLUG expression, as indicator of pathological advancement was determined using IHC staining. Significant increases of CAPN1/2-CAST-ERK1/2 levels of mRNA/protein were noted in SCLC compared to NCLM (p < 0.05). As a result, a higher level of CAPN1 and ERK1 genes was related to larger tumor size, more aggressive and deeper growth according to TFG scale and SLUG level (p < 0.05). There were also relationships of CAPN1/2 and ERK1 with incidences of local/nodal recurrences (p < 0.05). An inverse association for CAPN1/2, CAST, and ERK1/2 transcripts was determined with regard to overall survival (p < 0.05). In addition, a higher CAPN1 and phospho-ERK1 protein level was related to higher grade and stage (p < 0.05) and was found to promote worse prognosis. This is the first study to show that activity of CAPN1/2- CAST-ERK1/2 axis may be an indicator of tumor phenotype and unfavorable outcome in SCLC.

The effect of metallothionein 2A core promoter region single-nucleotide polymorphism on accumulation of toxic metals in sinonasal inverted papilloma tissues.

Metallothioneins (MTs) are intracellular thiol-rich heavy metal-binding proteins which join trace metal ions protecting cells against heavy metal toxicity and regulate metal distribution and donation to various enzymes and transcription factors. The goal of this study was to identify the -5 A/G (rs28366003) single-nucleotide polymorphism (SNP) in the core promoter region of the MT2A gene, and to investigate its effect on allele-specific gene expression and Cd, Zn, Cu and Ni content in sinonasal inverted papilloma tissue (IP), with non-cancerous sinonasal mucosa (NCM) as a control. The MT2A promoter region -5 A/G SNP was identified by restriction fragment length polymorphism using 117 IP and 132 NCM. MT2A gene analysis was performed by quantitative real-time PCR. Metal levels were analyzed by flame atomic absorption spectrometry. The frequency of A allele carriage was 99.2% and 100% in IP and NCM, respectively. The G allele carriage was detected in 23.9% of IP and in 12.1% of the NCM samples. As a result, a significant association of -5 A/G SNP in MT2A gene with mRNA expression in both groups was determined. A significant association was identified between the -5 A/G SNP in the MT2A gene with mRNA expression in both groups. A highly significant association was detected between the rs28366003 genotype and Cd and Zn content in IP. Furthermore, significant differences were identified between A/A and A/G genotype with regard to the type of metal contaminant. The Spearman rank correlation results showed the MT2A gene expression and both Cd and Cu levels were negatively correlated. The results obtained in this study suggest that the -5 A/G SNP in the MT2A gene may have an effect on allele-specific gene expression and toxic metal accumulation in sinonasal inverted papilloma.

Gene and protein expression of glucose transporter 1 and glucose transporter 3 in human laryngeal cancer-the relationship with regulatory hypoxia-inducible factor-1α expression, tumor invasiveness, and patient prognosis.

Increased glucose uptake mediated by glucose transporters and reliance on glycolysis are common features of malignant cells. Hypoxia-inducible factor-1α supports the adaptation of hypoxic cells by inducing genes related to glucose metabolism. The contribution of glucose transporter (GLUT) and hypoxia-inducible factor-1α (HIF-1α) activity to tumor behavior and their prognostic value in head and neck cancers remains unclear. The aim of this study was to examine the predictive value of GLUT1, GLUT3, and HIF-1α messenger RNA (mRNA)/protein expression as markers of tumor aggressiveness and prognosis in laryngeal cancer. The level of hypoxia/metabolic marker genes was determined in 106 squamous cell laryngeal cancer (SCC) and 73 noncancerous matched mucosa (NCM) controls using quantitative real-time PCR. The related protein levels were analyzed by Western blot. Positive expression of SLC2A1, SLC2A3, and HIF-1α genes was noted in 83.9, 82.1, and 71.7% of SCC specimens and in 34.4, 59.4, and 62.5% of laryngeal cancer samples. Higher levels of mRNA/protein for GLUT1 and HIF-1α were noted in SCC compared to NCM (p < 0.05). SLC2A1 was found to have a positive relationship with grade, tumor front grading (TFG) score, and depth and mode of invasion (p < 0.05). SLC2A3 was related to grade and invasion type (p < 0.05). There were also relationships of HIF-1α with pTNM, TFG scale, invasion depth and mode, tumor recurrences, and overall survival (p < 0.05). In addition, more advanced tumors were found to be more likely to demonstrate positive expression of these proteins. In conclusion, the hypoxia/metabolic markers studied could be used as molecular markers of tumor invasiveness in laryngeal cancer.

Metallothionein 2A core promoter region genetic polymorphism and its impact on the risk, tumor behavior, and recurrences of sinonasal inverted papilloma (Schneiderian papilloma).

Inverted papillomas are a unique group of locally aggressive benign epithelial neoplasms in the nasal cavity and paranasal sinuses arising from the Schneiderian mucosa. Metallothioneins are sulfhydryl-rich heavy metal-binding proteins required for metal toxicity protection and regulation of biological mechanisms including proliferation and invasion. The goal of this study was to identify three SNPs at loci -5 A/G (rs28366003) and -209 A/G (rs1610216) in the core promoter region and at locus +838 C/G (rs10636) in 3'UTR region of the MT2A gene with IP risk and with tumor invasiveness according to Krouse staging. Genotyping was performed using the PCR restriction fragment length polymorphism technique in 130 genetically unrelated IP individuals, and 418 randomly selected healthy volunteers. The presence of the rs28366003 SNP was significantly related to the risk of IP within the present population-based case-control study. Compared to homozygous common allele carriers, heterozygosity and homozygosity for the G variant had a significantly increased risk of IP (adjusted odds ratio [OR] = 7.71, 95% confidence interval [CI]: 4.01-14.91, p(dominant) < 0.001). Moreover, risk allele carriers demonstrated higher Krouse stage (pT1 vs. pT2-4) (OR = 19.32; 95% CI, 2.30-173.53; p < 0.0001), diffuse tumor growth (OR = 4.58; 95% CI, 1.70-12.11; p = 0.0008), bone destruction (OR = 4.13; 95% CI, 1.50-11.60; p = 0.003), and higher incidence of tumor recurrences (OR = 5.11; 95% CI, 1.68-15.20; p = 0.001). The findings suggest that MT2A gene variation rs28366003 may be implicated in the etiology of sinonasal inverted papilloma in a Polish population.

Expression of Th17 cell population regulatory cytokines in laryngeal carcinoma - Preliminary study.

AIM OF THE STUDY: Aim of the study was to evaluate the potential role of regulatory and proinflammatory cytokines IL-23 and IL-17 as Th17 lymphocyte activity markers in relation to invasiveness in laryngeal cancer. MATERIAL AND METHODS: The immunological analysis was conducted in 50 patients treated for squamous cell laryngeal carcinoma and 30 healthy volunteers as controls. The levels of IL-23 and IL-17 in supernatants of purified peripheral blood mononuclear cell cultures were determined by using the enzyme-linked immunosorbent assay (ELISA). The clinicomorphological criteria included pTNM, stage, G, and the total tumour front grading (TFG) score. RESULTS: Our data demonstrated higher concentrations of IL-23 in patients as compared to controls (p = 0.0001). No statistical difference for IL-17 in these groups was observed. Our study revealed significant dependences in IL-23 expression on pT (p = 0.04), histological differentiation (p = 0.04), and TFG total score (p = 0.02). Advanced tumours (pT3-pT4) with higher grade (G2-G3) and higher invasiveness (> 14 TFG points) were characterised by elevated IL-23 levels in PBMC supernatants. Our data did not indicate a relationship between cytokine levels and three- and five-year survival. However, a tendency towards lower content of IL-23 in PBMC cultures in patients who lived longer than five years after treatment was noted. The relationships between IL-17 level in PBMC cultures and clinicomorphological and prognostic parameters have not been disclosed. CONCLUSIONS: The results of this study suggest the importance of regulatory cytokine IL-23 in determining the aggressive potential of laryngeal carcinomas.

Expression of prostaglandin E2 prostanoid receptor EP2 and interleukin-1ß in laryngeal carcinoma - preliminary study.

AIM OF THE STUDY: Expression of EP2 protein, the prostaglandin E2 (PGE2) receptor, produced by tumour microenvironment inflammatory cells as well as tumour cells, may promote cellular proliferation and growth in an autocrine and paracrine fashion. The phenomenon involving these proteins is regulated by interleukin 1ß (IL-1ß). Many researchers indicate a connection of EP2 and IL-1ß in various types of neoplasms with higher tumour progression and poor prognosis. The aim of this study was to analyse the EP2 expression within laryngeal carcinoma tissue and IL-1ß levels in peripheral blood mononuclear cell supernatants and to find relationships between clinicomorphological features. MATERIAL AND METHODS: A group of 50 patients with verified squamous cell laryngeal carcinoma was analysed in this study. The pathological evaluation included pTNM depth of invasion according to tumour front grading criteria. Immunohistochemical analysis for membranous staining of EP2 in tumour tissues was used. The IL-1ß expression was determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: Increased EP2 expression in carcinoma cells was confirmed for more advanced tumours (pT3-pT4 vs. pT1-pT2, p < 0.0001 and pN1-3 vs. pN0, p = 0.02). Tumours with the highest aggressiveness identified by deeper invasion of submucosa or cartilage were characterised by the highest expression of EP2 (p < 0.0001). In laryngeal carcinomas characterised by a lower differentiation the highest EP2 expression in tumour cells was noted (p = 0.009). A positive relationship between IL-1ß expression and the presence of lymph node metastases was also confirmed (p = 0.04). CONCLUSIONS: The study indicates the potential effect of EP2 receptor and IL-1ß on tumour progression in laryngeal carcinoma.

The -5 A/G single-nucleotide polymorphism in the core promoter region of MT2A and its effect on allele-specific gene expression and Cd, Zn and Cu levels in laryngeal cancer.

Metallothioneins (MTs) are low molecular weight, cysteine-rich heavy metal-binding proteins which participate in the mechanisms of Zn homeostasis, and protect against toxic metals. MTs contain metal-thiolate cluster groups and suppress metal toxicity by binding to them. The aim of this study was to determine the -5 A/G (rs28366003) single-nucleotide polymorphism (SNP) in the core promoter region of the MT2A gene and to investigate its effect on allele-specific gene expression and Cd, Zn and Cu content in squamous cell laryngeal cancer (SCC) and non-cancerous laryngeal mucosa (NCM) as a control. The MT2A promoter region -5 A/G SNP was determined by restriction fragment length polymorphism using 323 SCC and 116 NCM. MT2A gene analysis was performed by quantitative real-time PCR. The frequency of A allele carriage was 94.2% and 91.8% in SCC and NCM, respectively, while G allele carriage was detected in 5.8% and 8.2% of SCC and NCM samples, respectively. As a result, a significant association was identified between the -5 A/G SNP in the MT2A gene with mRNA expression in both groups. Metal levels were analyzed by flame atomic absorption spectrometry. The significant differences were identified between A/A and both the A/G and G/G genotypes, with regard to the concentration of the contaminating metal. The Spearman rank correlation results showed that the MT2A expression and Cd, Zn, Cu levels were negatively correlated. Results obtained in this study suggest that -5 A/G SNP in MT2A gene may have an effect on allele-specific gene expression and accumulation of metal levels in laryngeal cancer.

Expression of cell adhesion molecules in laryngeal carcinoma - preliminary analysis.

AIM OF THE STUDY: Intercellular adhesion molecules present in immunocompetent cells as well as endothelium and tumour cells can regulate cell migration, angiogenesis, apoptosis, proliferation, and metastases in solid tumours. The aim of this study was to analyse the sICAM-1 (soluble intercellular adhesion molecule 1) and sVCAM-1 (soluble vascular cell adhesion molecule 1) expression in peripheral blood mononuclear cell (PBMC) cultures, and to find their relationships with clinicomorphological characteristics in laryngeal cancer. MATERIALS AND METHODS: The analysis included a group of 50 patients with verified squamous cell carcinoma of the larynx. The control group constituted 30 healthy volunteers. The pathological assessment included pTNM, stage, histological grade, and type of invasion according to the tumour front grading. The expression of adhesion molecules was assessed using the enzyme-linked immunosorbent assay (ELISA). RESULTS: Increased expression of sICAM-1 and sVCAM-1 was an indicator of more aggressive laryngeal carcinomas. More advanced local changes evaluated on the pT feature were connected with a higher sVCAM-1 (p = 0.017), but not sICAM-1 level. The presence of lymph node metastases correlated with a higher expression of adhesion molecules (p = 0.012 and p = 0.003, for sICAM-1 and sVCAM-1, respectively). Tumours with more diffuse growth and infiltrating with small cell groups (< 15/hpf) was characterised by the highest level of adhesive proteins (p = 0.001 and p = 0.02 for sICAM and sVCAM, respectively). Moreover, lower levels of sICAM-1 and sVCAM-1 were observed more frequently in patients who lived longer than five years after treatment. CONCLUSIONS: The study indicates the importance of the sICAM and sVCAM expression as indicators of advanced changes and prognosis in patients with laryngeal carcinoma.

Diagnostic impact of promoter methylation and E-cadherin gene and protein expression levels in laryngeal carcinoma.

AIM OF THE STUDY: Inactivation of the tumor suppressor E-cadherin (CDH1) and its decreased expression is an important occurrence during carcinogenesis. Nevertheless, the relationship of CDH1 expression and the promoter methylation with laryngeal cancer cell aggressiveness is still unclear. The purpose of this study was to elucidate the gene and protein E-cadherin expression and the DNA methylation levels and to describe the correlations with morphological features in squamous cell laryngeal cancer. MATERIAL AND METHODS: The authors studied E-cadherin and the DNA methylation level in 86 cases to gain a further understanding of the clinicopathologic significance of analyzed parameters. The pathological evaluation included pTNM classification and the tumor front grading (TFG) criteria. Quantitative analysis of the amplified product in real time (qRT-PCR) for estimation of CDH1 mRNA was used. The methylation status was investigated by using methyl-specific polymerase chain reaction (MSP). The level of CDH1 protein expression by Western blot was determined. RESULTS: Downregulation of E-cadherin was found to be related to promoter methylation (p < 0.001). In tumors with the highest invasiveness according to TFG criteria the lowest E-cadherin gene and protein level in the study group was observed (p = 0.046 and p = 0.0002, respectively). In SCLC with muscle and cartilage invasion and disperse infiltration the lowest CDH1 gene and protein expression was noted (p = 0.0003 and p = 0.003 for deep invasion, p = 0.033 and p = 0.003 for multifocal infiltration, respectively). CONCLUSIONS: The current findings suggest an association of E-cadherin tumor expression with progression of laryngeal cancer. CDH1 gene level may be an auxiliary molecular marker for advanced cases of laryngeal carcinoma; however, further studies are necessary.

Expression of CTLA-4 and Foxp3 in peripheral blood T cells of patients with squamous cell laryngeal carcinoma.

Cytotoxic T lymphocyte-associated antigen 4 (CTLA-4, CD152) and Foxp3 (forkhead box P3) are receptors present on T cells which play a critical role in the down-regulation of antigen-activated immune responses. To evaluate the potential influences of CTLA-4 and Foxp3 on cancer invasiveness, a case-control study was conducted in 86 patients treated for squamous cell laryngeal carcinoma. The abundance of CTLA-4 and Foxp3 gene transcripts in the purified peripheral blood mononuclear cells (PBMCs) by quantitative real-time PCR (qRT-PCR) was determined. The analysis of proteins by Western blot was performed. The relationships between CTLA-4 and Foxp3 gene and protein expression as well as the aggressiveness of tumor determined on pT, type and depth of invasion were investigated. Our work revealed a significant dependence of mRNA CTLA-4 on tumor front grading (TFG) total score (p = 0.04) as well as CTLA-4 protein expression on pT (p = = 0.03) and type of invasion (p = 0.03). Advanced pT3-pT4 tumors with diffuse infiltration and > 14 TFG points were characterized by higher average values of CTLA-4 protein in PBMCs. Our data also demonstrated significant differences between Foxp3 protein levels in relation to pT (p = 0.04), depth of invasion (p = = 0.02) and type of invasion (p = 0.03). In tumors with the highest invasiveness identified by the pT3-pT4 status, deep invasion with involvement of cartilage and diffuse infiltration, the highest Foxp3 protein level was observed. In conclusion, these results suggest an impact of CTLA-4 and Foxp3 in determining proliferative and aggressive potential of laryngeal carcinoma, highlighting the significance of CTLA-4 and Foxp3 as potential predictive indicators.

[The role of stromal mast cells in the modification of CD4+CD25+Foxp³ regulatory T cells, Th17 lymphocytes and cytotoxic lymphocytes Tc1 in the development and progression of tumor]. / Rola komórek tucznych naciekajacych guz w modyfikacji funkcji limfocytów regulatorowych CD4+CD25+Foxp³, limfocytów Th17 oraz limfocytów cytotoksycznych Tc1 w rozwoju i progresji guza nowotworowego.

Despite the lack of direct evidence that immune surveillance cells protect against tumor development, indirect clinical observations and experimental studies indicate activity in the immune response against cancer cells of various origin. Little is known about the effects of the stromal tumor mast cell (MC) in the activity of immune cells, i.e. CD4+CD25+Foxp³+ regulatory T cells, Th17 lymphocytes, cytotoxic lymphocytes Tc1 and their mutual modulatory function and regulation of the antitumor immune response. Factors synthesized by stromal tumor mast cells including histamine, COX-2, CXCL8 (IL-8), VEGF, IL-6, TNF, iNOS, MMP-8, and MMP-9 may, on the one hand, directly affect the activity of T lymphocyte subpopulations, i.e. iTreg, Tc1, and Th17, and thus regulate immunological processes occurring in the vicinity of the tumor. On the other hand, through effects on angiogenesis, apoptosis, the cell cycle, secretion of cytokines and the expression of adhesion molecules, they may indirectly determine the progression of the neoplasm. Understanding the regulatory mechanisms occurring in the system: tumor stroma mast cell → immune cells infiltrating the tumor (iTreg, Tc1, Th17 lymphocytes) → expression of factors involved in angiogenesis, apoptosis, the cell cycle, and secretion of cytokines and adhesion molecules creates the future possibility of influencing the activation and regulation of selected proneoplastic and antineoplastic factors appearing in the neoplasm environment. Research on these mechanisms may be the beginning of a new approach to the fight against cancer growth and provide an opportunity to introduce new methods of treatment. The aim of this study was to present the current knowledge on the role of stromal tumor CD117+ mast cells and factors secreted by these cells in the activation of T lymphocyte subpopulations, i.e. CD4+CD25+Foxp³+ regulatory T cells, Th17 lymphocytes, and cytotoxic lymphocytes Tc1, as well as to present their impact on the degree of tumor invasiveness by regulating the synthesis of factors secreted by the lymphocyte subpopulations studied, e.g. IL-10 and TGF-ß (iTreg), IL-17A and IL-6 (Th17), IFN-γ and IL-2 (Tc1).

Impact of EGFR immunoexpression on STAT3 activation and association with proinflammatory/regulatory cytokine pattern in laryngeal squamous cell carcinoma.

Stimulation of epidermal growth factor receptor (EGFR) results in the activation of signal transducer and activator of transcription-3 (STAT3), a transcriptional factor associated with carcinogenesis. Proinflammatory cytokines are capable of activating a tumor cell death program by reducing EGFR tyrosine phosphorylation. This study aimed to identify EGFR expression in laryngeal carcinoma and determine the relationship with STAT3 and proinflammatory/regulatory cytokine secretion. An analysis of EGFR expression (membranous EGFR-m and cytoplasmic EGFR-c) was performed in tumor tissues by immunohistochemical (IHC) staining in 45 medical cases of laryngeal carcinoma. STAT3 expression in freshly isolated tumor and non-cancerous normal epithelial cells by RT-PCR was analyzed in 24 patients after total larynx resection. The concentrations of TNFalpha, IL-2, IL-6, IL-8 and IFNgamma secreted by purified peripheral blood mononuclear cells (PBMCs) or contained in whole blood samples were measured by ELISA. The relationship between EGFR and mRNA STAT3 expression as well as the level of secreted cytokines was investigated. In our study, 93.3% tumors expressed EGFR-m and 37.8% EGFR-c. It also revealed a statistically significant dependence of the EGFR status on STAT3 expression in neoplastic tissues. Tumors with IHC EGFR-m positive staining >50% of the total number of cells, as well as with EGFR-c positive staining, were characterized by the most frequent presence of STAT3 expression. Our data demonstrate a significant negative relationship between EGFR-m expression and TNFalpha concentration, and a positive connection between membranous EGFR and IL-8 or IFNgamma levels recorded in isolated PBMCs. Furthermore, this study revealed a significant relationship between EGFR-c immunoexpression and IL-8 or IFNgamma concentration. Our findings have confirmed a key role of EGFR in determining the proliferative and malignant potential of laryngeal carcinoma.

EGFR immunoexpression and peripheral blood cytokine secretion as potential biomarkers of tumor behavior in laryngeal squamous cell carcinoma.

BACKGROUND: Epidermal growth factor receptor (EGFR) and proinflammatory cytokines have been implicated in the dysregulated cell growth and increased aggressiveness of human cancers. The purpose of this study was to analyze EGFR immunoexpression in neoplastic tissues and IL-6 and TNFalpha secretion by peripheral blood mononuclear cells (PBMCs) and to investigate their relationships with certain clinicopathological characteristics. MATERIAL/METHODS: Tumor expression of membranous and cytoplasmic EGFR was measured in 45 cases of laryngeal squamous cell carcinoma by IHC staining. IL-6 and TNFalpha concentrations in 21-h PBMC cultures were measured by ELISA. Relationships between EGFR and cytokine secretion and clinicopathological characteristics such as pT status, pN status, and TFG classification, which include the parameters of the most invasive zones of neoplastic tissue and histological grade, were analyzed. RESULTS: The membranous EGFR index had a very strong association with pT stage, mode of invasion, and lymphocytic plasma infiltration of the tumor stroma. Relationships between the cytoplasmic EGFR index and nuclear polymorphism as well as TFG score for advanced carcinomas and histological grade in less invasive tumors were highlighted. The correlations of IL-6 and TNFalpha levels with TFG score, pT status, histological grade, and mode of tumor invasion were also significant. CONCLUSIONS: These findings confirmed the importance of EGFR immunoexpression rate as well as IL-6 and TNFalpha secretion by PBMCs as potential biomarkers for assessing the aggressive tumor phenotype in laryngeal carcinoma.

[Production of proinflammatory cytokines by peripheral blood mononuclear cells in cocultures with squamous cell carcinoma of the larynx]. / Wydzielanie cytokin prozapalnych przez jednojadrzaste komórki krwi obwodowej w hodowli z komórkami raka plaskonablonkowego krtani.

PURPOSE: Antitumor mechanisms of cellular immune response are connected with the cytokines activity secreted by peripheral blood mononuclear cells (PBMC). The aim of this study was estimation of proinflammatory cytokine (IL-6 and TNF- ) concentration in patients with laryngeal squamous cell carcinoma and analysis of directed influence of neoplasm cells to the function of PBMC and modification of cytokine profile. MATERIALS AND METHODS: For analysis of cytokine secretion, the cultures of isolated peripheral blood mononuclear cells (PBMC) with marginal neoplasm cells and noncancerous adjacent mucosa cells from 55 patients with laryngeal squamous cell carcinoma were established. Production of cytokines in 21h and 72h culture supernatants was detected by Elisa. RESULTS: Authors reported the increase of IL-6 and decrease of TNF- concentration in cultures of isolated peripheral blood mononuclear cells (PBMC) with marginal neoplasm cells and non-cancerous adjacent mucosa cells. CONCLUSION: The presence of laryngeal squamous carcinoma cells in PBMC culture can modify immunocompetent cells activity and production of proinflammatory cytokines.

[Expression of transcription nuclear factor NFkappaB in laryngeal carcinoma tumor cells--correlation with IL-10 expression by blood mononuclear cells and clinicomorphological features]. / Ekspresja transkrypcyjnego czynnika jadrowego NFkappaB w komórkach raka krtani--korelacja z ekspresja IL-10 oraz cechami kliniczno-morfologicznymi guza.

PURPOSE: Transcription nuclear factor NFkappaB (p65-p50/RelA) is an activator of transcription process and regulates the apoptosis, cell cycle, proliferation, secretion of cytokines, angiogenic and growth factors by initiation gene transcription. NFkappaB can be activated by signaling pathway following IL-10 stimulation. The aim of this study was to estimate NFkappaB cytoplasmic and nuclear immunoexpression and to analyze the connection with clinicopathological features and IL-10 concentration produced by blood mononuclear cells in patients with laryngeal carcinoma. MATERIALS AND METHODS: Analysis of NFkappaB expression by immunmohistochemistry and IL-10 production by PBMCs and measured by Elisa in 45 patients with squamous cell carcinoma of the larynx were performed. Connections NFkappaB immunoexpression with clinicomorphological features (pT, pN, Anneroth, Batsakis i Lunas' classification) and IL-10 secretion were analyzed. RESULTS: Authors reported statistical correlation between NFkappaB cytoplasmic level and clinicomorphological parameters such as neoplasm advance according to Anneroth, Batsakis i Lunas' classification and depth of invasion as well as IL-10 secretion by PBMCs. CONCLUSION: Our studied indicated the important influence of NFkappaB activity on advance in laryngeal carcinoma as well as connection of anti-inflammatory and regulatory cytokine IL-10 produced by immunocompetent cells for course of neoplasm disease.

[Clinical ocular-motor disturbances in multiple sclerosis]. / Kliniczna ocena zaburzen ruchowych galek ocznych w stwardnieniu rozsianym.

UNLABELLED: Otoneurologic bedside examination with testing eye movements gives valuable information about static and dynamic properties of balance system and may give topodiagnostyc information about the side of lesion in patients with vertigo, dizziness and disequilibrium. THE AIM OF THE STUDY: was to present the scheme of otoneurological bedside examination and usefulness of ocular motor disturbances index in evaluation of Multiple Sclerosis patients status. MATERIAL AND METHODS: Sixty patients with diagnosis of MS, seen in outpatient neurology clinic, Medical University of Lodz, from 2002 to 2004, were enrolled into the study. Patient's history of vertigo, dizziness, hearing loss and vision disturbances were evaluated. The clinical bedside ocularmotor examination was performed in all patients. It was composed of seven tests on the basis on which we introduce ocular motor disturbances index--IRZ. RESULTS: The most frequent abnormalities were found in clinical saccadic test in 30% and smooth pursuit in 22%. MS patients who had in clinical eye movements examination IRZ bigger than 3 point formed the abnormal clinical examination group' (ACE)-- 31.7%. In u 68.3% the index was less than normal clinical examination group' (NCE). The longer duration of the disease was observed in ACE group. Comparisons of functional neurological score EDSS and for cerebellar and brainstem subscale were significantly greater in ACE group. CONCLUSION: Otoneurological bedside examinations with dynamic tests and introducing quantitative the ocular motor disturbances index is a valuable method of evaluation of visual-ocularmotor reflex and may be used in monitoring MS course of disease.

[Expression of activation antigens on the lymphocytes T in patients with carcinoma of the larynx and connection with tumor features]. / Ekspresja wybranych antygenów aktywacji na limfocytach T u chorych z rakiem krtani a cechy kliniczno-morfologiczne guza.

INTRODUCTION: Results of studies analyzing the role of immunocompetent cells in tumor environment and whole peripheral blood indicate their responsibility for aggressiveness of neoplasm, prognosis and therapeutic effect. Atcivation of lymhocytes T is connected with expression the markers (antigens) on their surface. The aim of this study was the analysis of activation antigens expression on lymphocytes T in patients with laryngeal carcinoma and the connection with clinicomorphological features. MATERIAL AND METHODS: Analysis of activation antigens expression CD69, CD71 and CD25, CD26, HLA/DR on lymphocytes T CD4+ i CD8+ in 33 patients with squamous cell carcinoma of the larynx was performed. Flow cytometry-based analysis of activation antigens in T cell cultures with and without PHA stimulation was used. The connection of these molecules and clinicomorphological features was examined (pT, pN, G, Anneroth, Batsakis and Lunas' classification). RESULTS: The significant correlation between chosen markers of activation and tumor features were noted: pT with HLA/DR/CD4, CD69CD8, CD71CD8, pN with CD26CD8, G with CD25CD8, CD71CD8, ABL score with CD25CD4. CONCLUSION: Our data indicated the connetion of immunocompetent cell activity and spread of neoplasm in patients with laryngeal carcinoma.

[Results of treatment papilloma of nasal cavity and paranasal sinuses]. / Wyniki leczenia brodawczaków jamy nosa i zatok przynosowych.

INTRODUCTION: Papilloma of the nose and paranasal sinuses is a benign tumor originated from nose mucosa. Especially inverted papilloma tumor has a significant recurrence and malignancy potential rate. The aim of the study was the analysis of clinical and treatment outcomes of patients with papilloma of the nose and paranasal sinuses. MATERIAL AND METHODS: The retrospective analysis was curried out on 41 patients--16 with papiloma of the nasal vestibule and 25 with inverted papilloma the nose and paranasal sinuses surgically treated in I ENT Clinic Medical University in Lodz between 1998-2004 years. We analyzed patient's complains, clinical data and surgical follow-up results. RESULTS: The most frequent complains was increasing unilateral nasal obstruction and rhinorhea. Nasal vestibule papilloma were intranasal removed in all cases. In extended tumor nose and paranasal sinuses in 14 cases intranasal procedures, in 7 sublabial approached, in 4 lateral rhynothomy were performed. In 5 patient local recurrences was observed and in 3 neoplasmatic transformation. CONCLUSION: The choice of surgical management should be individual with respect to tumor localization and extension of neoplasmatic process. The treatment result depends of radical tumor resection.