TC2 C776G polymorphism studies in patients with oral cancer in the Polish population.

The first signs of oral cancer may resemble developing infections in the mucous membranes, with throat cancer symptoms being similar to those of upper respiratory tract infections. This greatly hinders rapid diagnosis and treatment. Better knowledge of the changes occurring in the metabolism of folic acid can help in understanding the carcinogenesis affecting DNA methylation and genome stability. Polymorphisms in genes encoding enzymes involved in this pathway may influence enzyme activity and thereby interfere with the concentrations of homocysteine and S-adenosylmethionine, which are important for DNA synthesis and cellular methylation reactions. The aim of the study was to determine the risk of oral cancer associated with the TC2 C776G polymorphism, as determined in 119 patients. Genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). The test genotype was found to correspond to the Hardy-Weinberg (HW) equilibrium (p > 0.05). In our population G/G homozygosity of C776G TC2 gene polymorphism increases the risk of oral cancer; OR (odds ratio): 4.3875; 95% CI (confidence interval): 2.0518-9.319; p = 0.001. Regarding C/G genotype of the C776G TC2 gene, polymorphism also increases the risk of developing this cancer; OR 2.4146 95% CI: 1.2803-4.5541; p = 0.01.

Genetic variability of the endangered fish lake minnow (Eupallasella percnurus) in populations newly established by translocation and those existing long term in Poland.

The lake minnow Eupallasella percnurus is a small leuciscid fish. In Poland, this species has been in a continuous decline since the mid-20th century and is presently considered as a extremely endangered. According to Polish law, E. percnurus is a strictly protected species that requires active conservation measures. In Poland, one the most common and effective measure of active protection E. percnurus is initiation of new populations. For this purpose, in 2004-2012, juvenile individuals originating from aquaculture conditions were translocated to group of isolated water bodies not inhabited by this species. The juveniles were offspring of parental fish belonging to the same local population, which is extinct at present. Five of those attempts were successful. The aim of the present study was to assess the genetic variation in a group new populations and compare genetic variation indicators with 13 old populations that had existed for decades. The polymorphism of 13 microsatellite markers was investigated, significance of differences in the genetic variation indicators between the groups were tested using a one-way analysis of variance (ANOVA). The mean values of all summary statistics under study, i.e. observed heterozygosity, expected heterozygosity and the total number of alleles, were higher in the group of new populations compared to almost all old ones. A similar dependence was found for Garza-Williamson M values, where the mean for the group of new populations was higher than in almost all old populations. Our results indicate that all recently established E. percnurus populations have not yet experienced any extensive founder effects or bottlenecks. They have preserved a large part of the genetic variability typical of their maternal population, which might also have been relatively high. This feature of new populations, may give them a relatively high ability to adapt to changing environments in the future.

Ribosomal intergenic spacer analysis as a tool for monitoring methanogenic Archaea changes in an anaerobic digester.

The applicability of a newly-designed PCR primer pair in examination of methanogenic Archaea in a digester treating plant biomass was evaluated by Ribosmal Intergenic Spacer Analysis (RISA). To find a suitable approach, three variants of RISA were tested: (1) standard, polyacrylamide gel-based, (2) automated, utilized capillary electrophoresis (GA-ARISA), and (3) automated microfluidics-based (MF-ARISA). All three techniques yielded a consistent picture of archaeal community structure changes during anaerobic digestion monitored for more than 6 weeks. While automated variants were more practical for handling and rapid analysis of methanogenic Archaea, the gel-based technique was advantageous when micro-organism identification was required. A DNA-sequence analysis of dominant bands extracted from the gel revealed that the main role in methane synthesis was played by micro-organisms affiliated with Methanosarcina barkeri. The obtained results revealed that RISA is a robust method allowing for detailed analysis of archaeal community structure during organic biomass conversion into biogas. In addition, our results showed that GA-ARISA has a higher resolution and reproducibility than other variants of RISA and could be used as a technique for tracking changes in methanogenic Archaea in an anaerobic digester.

Management of Genetic Variation in the Gamete Bank of the Endangered Lake Minnow Eupallasella percnurus, Using Genassemblage 2.2 Software.

The management of genetic resources deposited in gene banks requires knowledge of the genetic profiles of the gamete donors and bioinformatics tools to process this information. In this work, we show how to use Genassemblage 2.2 software in managing the genetic variation deposited in a bank of cryopreserved semen. Our demonstration was based on the leuciscid fish species, lake minnow Eupallasella percnurus, which is designated as endangered in Poland. The semen samples (n = 192) were taken from four Polish lake minnow populations and frozen in the gene bank. Fin clips were taken and DNA extracted. Across 13 investigated microsatellite loci, 21-53 alleles were identified in each population and 66 in the entire group of samples. The module "Management of genetic variation in gamete bank" of Genassemblage 2.2 software was used to find the set of samples that will preserve 100% of the detected allelic diversity in the next generation. Our results showed that a small group of 4-19 semen samples was enough to transfer all alleles detected across this set of samples. We, therefore, recommend Genassemblage 2.2 as a convenient tool for the detection of genetic differences between donors, the construction of optimal sets of samples for conservation of genetic variation, and for managing genetic variation deposited in gamete banks. Consequently, it can be used in breeding human-dependent populations and to optimize the use of genetic diversity in samples in the gamete banks. It can be especially useful for conserving populations of species characterized by low genetic variation, such as the lake minnow.

Cytotoxicity and genotoxicity of capecitabine in head and neck cancer and normal cells.

The interaction between a chemical and a cell may strongly depend on whether this cell is normal or pathological. Side effects of anticancer drugs may sometimes overcome their benefit action, so it is important to investigate their effect in both the target and normal cells. Capecitabine (Xeloda, CAP), a prodrug of 5-fluorouracil, is mainly used in colon cancer, but little is known about its action in head and neck cancer. We compared the cyto- and genotoxicity of CAP in head and neck HTB-43 cells and normal human lymphocytes by comet assay and flow cytometry. CAP at concentration up to 50 µM significantly decreased the viability of the cancer cells, whereas it did not affect normal lymphocytes. The drug did not interact with isolated plasmid DNA, but it damaged DNA in both cancer and normal cells. However, the extent of the damage in the former was much higher than in the latter. CAP induced apoptosis in the cancer cells, but not in normal lymphocytes. Pre-treatment of the cells with the nitrone spin traps α-(4-pyridil-1-oxide)-N-tert-butylnitrone and N-tert-butyl-α-phenylnitrone decreased the extent of CAP induced DNA damage, suggesting that free radicals may be involved in the formation of DNA lesions induced by CAP. The drug evoked an increase in the G0/G1 cell population accompanied by a decrease in the S cell population. CAP may evoke a pronounced cyto- and genotoxic effects in head and neck cancer cells, whereas it may or may not induce such effects in normal cells to far lesser extent.

Difficulties associated with the diagnosis of mycosis of the oral cavity and throat in chronic lymphocytic leukemia (CLL).

Cases of fungal infections are being encountered more often in clinical practice. The factors associated with a high risk of mycoses include, among others, corticosteroidotherapy, the administration antibiotics with wide spectrum of antibacterial properties, neutropenia, neoplasms. Fungi may play a role in cancer formation, may act as a complication in the course of treatment, and may mimic a neoplastic process by giving a similar clinical picture. In the case of fungal throat infection, patients complain of increased body temperature, a general feeling of weakness, malaise, headache, spontaneous pain intensifying during swallowing, a feeling of an obstacle in the throat or a cough. A physical examination may reveal congestion of the mucosa followed by a unilateral crater ulceration often covered with fat, as well as a thick coating, which is accompanied by foetor ex ore. The submandibular and neck lymph nodes are often greatly enlarged and painful. These symptoms may resemble those associated with the neoplastic process and changes in the course of systemic diseases (agranulocytosis). A correct diagnosis in these cases is necessary for adequate therapy. Chronic lymphocytic leukemia (CLL) is the most common type of leukemia among adults in Europe and North America. It is estimated that in Poland, CLL affects approximately 1,400 people per year. In this paper, a case of 62-years old patient with CLL with fungal infection of oral cavity and throat is presented.

The human aural myiasis caused by Lucilia sericata.

Myiasis is a rare, worldwide, human disease with seasonal variation, caused by developing larvae of a variety of fly species. It can be dangerous when infestations penetrate into the brain. In the available literature, we have found only a few papers concerning ear myiasis caused by Lucilia sericata. Here, we report 2 cases of aural myiasis. Early intervention (surgical removal, occlusion) in these cases should prevent complications. Larvae, for further examination, should be killed by immersion in very hot water, then preserved in an ethanol.

Influence of Arg399Gln, Arg280His and Arg194Trp XRCC1 gene polymorphisms of Base Excision Repair pathway on the level of 8-oxo-guanine and risk of head and neck cancer in the Polish population.

BACKGROUND: Reduced efficiency of DNA repair systems has long been a suspected factor in increasing the risk of cancer. OBJECTIVE: In this work we investigate influence of three selected polymorphisms of DNA repair gene XRCC1 and level of oxidative damage (measured as level of 8-oxo-guanine) on modulation of the risk of HNSCC. METHODS: In group of 359 patients with HNSCC (diagnosed with OSCC) the occurrence of polymorphic variants in Arg399Gln, Arg280His and Arg194Trp of XRCC1 were studied with TaqMan technique. In addition we determined level of 8-oxo-guanine with ELISA. RESULTS: Arg399Gln polymorphism and Arg194Trp polymorphism of XRCC1 gene increases the risk of HNSCC. The coexistence of Arg399Gln and Arg194Trp simultaneously enhances this effect. At the same time, their coexistence with His280His raises the risk to a level higher than in the absence of such coexistence, although the His280His itself is not associated with an increased risk of HNSCC. Patients have higher levels of 8-oxo-guanine than control group, and His280His is polymorphism with highest mean value of 8-oxoG level among studied. CONCLUSION: Patients with HNSCC not only have an increased level of 8-oxoguanine and the Arg399Gln and Arg/Trp of XRCC1 modulate risk of cancer, but there is also a relationship between these two phenomena, and it can be explained using intragenic combinations revealing that a high level of 8-oxoG could be a potential mechanism behind the modulation of HNSCC risk by the polymorphisms studied.

Genassemblage 2.0 software facilitates conservation of genetic variation of captively propagated species.

In conservation of captively propagated species, conserving genetic diversity is important. Here, we present an example of the use of Genassemblage 2.0 software in conserving the genetic variation of the lake minnow (Eupallasella percnurus). This fish has low genetic variation and is at risk of extinction in the western edge of its range, which includes Poland. Fish from one Polish population were captured (23 males, 25 females). Fin clips were taken, and DNA was extracted. Polymorphic microsatellites (13) were used to prepare genetic profiles, assess genetic variation in the fish and estimate genetic diversity in their progeny. Alleles were scored using an automatic capillary sequencer. The four and eight best variants of spawning pairs, and the optimal sets for group volitional breeding (four males, four females; eight males, eight females) were identified using Genassemblage 2.0. In the sets of 8 and 16 fish for group breeding, the mean heterozygosity, the number of alleles, and the share of "weak" heterozygotes (0.493, 24, 0.239 and 0.479, 23, 0.257, respectively) were better than the mean values for the progeny of all potential breeding pairs. For group volitional breeding, one set of four males and four females, and numerous sets of eight males and eight females would enable transmission of all 33 alleles identified in the potential broodstock and an expected progeny heterozygosity of 0.441 and 0.414, respectively. These expected heterozygosity values are higher than those in the broodstock. For practical purposes, the larger sets would be preferable for avoiding a future inbreeding and genetic drift.

Comparative study of DNA damage and repair in head and neck cancer after radiation treatment.

We compared DNA damage and the efficacy of its repair after genotoxic treatment with gamma-radiation of lymphocytes and tissue cells isolated from patients with squamous cell carcinoma of head and neck (HNSCC) and healthy donors. Thirty-seven subjects with HNSCC and 35 healthy donors were enrolled in the study. The extent of DNA damage including oxidative lesions and efficiency of the repair were examined by alkaline comet assay. HNSCC cancer cells were more sensitive to genotoxic treatment and displayed impaired DNA repair. In particular, lesions caused by gamma-radiation were repaired less effectively in metastasis of HNSCC than in healthy controls. The differences in radiation sensitivity of cancer and control cells suggested that DNA repair might be critical for HNSCC treatment. We conclude that gamma-radiation might be considered as an effective therapeutic strategy for head and neck cancers, including patients in advanced stage of the disease with clear evidence of metastasis.

Role of impaired DNA repair in genotoxic susceptibility of patients with head and neck cancer.

DNA repair is critical for genotoxic susceptibility and cancer development. Forty-seven patients with head and neck squamous cell carcinoma (HNSCC) and 38 healthy controls were enrolled in this study. Among the patients, 16 subjects had metastasis of HNSCC. The extent of DNA damage, including oxidative lesions, and efficiency of repair after genotoxic treatment with hydrogen peroxide were examined using the alkaline comet assay. HNSCC cells were sensitive to genotoxic treatment and displayed impaired DNA repair. In particular, lesions caused by hydrogen peroxide were repaired less effectively in cancer cells from patients with metastasis than in cells from healthy controls. We suggest that impaired DNA repair might play a role in genotoxic susceptibility of patients with head and neck cancer. Finally, as a consequence of this finding we have shown that treatment with DNA-reactive drugs could be considered as an effective therapy strategy for head and neck cancer.

The Cys326 allele of the 8-oxoguanine DNA N-glycosylase 1 gene as a risk factor in smoking- and drinking-associated larynx cancer.

Tobacco smoke-related products and ethanol would induce oxidative modifications to the DNA bases, thereby contributing to larynx cancer. Human 8-oxoguanine DNA N-glycosylase 1 (hOGG1) deals with oxidative DNA damage, and the base changes in the hOGG1 gene may alter the susceptibility of the human cells to tobacco smoke-related compounds and/or ethanol. In the present work, we investigated the association between smoking, drinking or the Ser326Cys polymorphism of the hOGG1 gene and the risk of larynx cancer in a Polish population. It has been reported that the Ser326 allele exhibits higher activity than the Cys326 variant. In this study, 253 age-matched controls and 253 patients with larynx cancer were enrolled. The polymorphism was determined with DNA from blood lymphocytes by polymerase chain reaction. The frequencies (%) of the genotypes were Ser/Ser 65.6, Ser/Cys 30.4, and Cys/Cys 4.0 in the controls and those in patients were 55.7, 36.0 and 8.3, respectively. Stratification of individuals according to their smoking and drinking habits indicated that these habits might be significant risk factors in larynx cancer. The Ser/Cys and Cys/Cys genotypes are significantly associated with the increased risk of larynx cancer. These genotypes increased the risk ratio of larynx cancer among heavy smokers, but did not change the risk in former smokers and moderate smokers. These genotypes also increased the risk of larynx cancer in moderate and heavy drinkers. Therefore, the Cys326 allele of the hOGG1 gene may increase the risk of larynx cancer associated with smoking or alcohol consumption.

DNA damage and repair of head and neck cancer cells after radio- and chemotherapy.

DNA repair is critical for successful chemo- and radiotherapy of human tumours, because their genotoxic sensitivity may vary in different types of cancer cells. In this study we have compared DNA damage and the efficiency of its repair after genotoxic treatment with hydrogen peroxide, cisplatin and gamma-radiation of head and neck squamous cell carcinoma (HNSCC). Lymphocytes and tissue cells from biopsies of 37 cancer patients and 35 healthy donors as well as the HTB-43 larynx cancer cell line were employed. The cell sensitivity to genotoxic treatment was estimated by the MTT survival assay. The extent of DNA damage and efficiency of its repair was examined by the alkaline comet assay. Among the examined treatments, we found that HNSCC cells were the most sensitive to gamma-radiation and displayed impaired DNA repair. In particular, DNA damage was repaired less effectively in cells from HNSCC metastasis than healthy controls. In conclusion, our results suggest that the different genotoxic sensitivity of HNSCC cells may depend on their DNA repair capacity what in turn may be connected with the effectiveness of head and neck cancer therapy.

Polymorphisms in the Bovine Tumour Necrosis Factor Receptor Type Two Gene (TNF-RII) and Cell Subpopulations Naturally Infected with Bovine Leukaemia Virus.

INTRODUCTION: Numerous mutations in the bovine tumour necrosis factor receptor type two (TNF-RII) gene have been identified, but their biological consequences remain poorly understood. The aim of this study was to determine whether polymorphism in the analysed loci of the bovine TNF-RII gene is linked with the size of cell subpopulations naturally infected with bovine leukaemia virus (BLV) which serve important immune functions in the host. MATERIAL AND METHODS: Samples originated from 78 cows. Polymorphisms in the studied gene were determined by PCR-RFLP and DNA sequencing by capillary electrophoresis. BLV infection was diagnosed by the immunofluorescence (IMF) technique and nested PCR. Cell subpopulations were immunophenotyped with IMF. RESULTS: Similar and non-significant differences in the average percentages of TNFα±, IgM+TNFα±, and CD11b+TNFα±cells infected with BLV were noted in individuals with various genotypes in the polymorphic sites g.-1646T > G and g. 16534T > C of the TNF-RII gene, and significant differences in the percentages of these subpopulations were observed between selected microsatellite genotypes (g.16512CA(n)). CONCLUSION: STR polymorphism and the number of CA dinucleotide repeats in intron 1 of the TNF-RII gene influence the frequency of TNF+, CD11b+TNF+, and IgM+TNF+ subpopulations naturally infected with BLV. Polymorphism in the gene's other two sites do not affect the size of these cell subpopulations.

Antioxidative assessment of new trans-palladium (II) complexes in head and neck cancer.

BACKGROUND: Head and neck neoplasms stand for 6% of all malignant neoplasms worldwide. Chemotherapy has limited use due to the biological properties of the tumor (in the majority of cases moderately and poorly differentiated squamous cell carcinoma). The fundamental molecule used in treatment is cisplatin and its derivates, that can be associated with fluorouracil. The new chemotherapeutic agents are not in common use during the treatment of head and neck malignancies. However, the use of low molecular weight complexes Pd (II) carries the potential of being more effective in therapy. MATERIAL AND METHODS: Fifty-one patients, 30 men and 21 women (aged 52.9 ± 12.1 years) with head and neck cancer were included in the study. Fifty-one healthy subjects, 31 men and 20 women, (aged 54.1 ± 14.7 years) years formed the control group. Antioxidant enzymes, superoxide dismutase, and catalase activities in erythrocytes were examined. RESULTS: An increased level of antioxidant enzymes was seen in the blood samples from patients with head and neck cancer after incubation with Pd (II) complex. In the group we obtained a statistically significant result p = <0.001. DISCUSSION: That project may contribute to the development of new, more efficient head and neck cancer treatment strategies. In our opinion, the results can be used in the future to develop a valuable prognostic marker of the disease. This is important because the initial phase of cancer is asymptomatic. The search for factors involved in pathogenesis translates into economic benefits and makes therapy more effectiveness through the reduction of treatment expenses.

[Use of tissue autofluorescence in the diagnostics and assessment of treatment efficacy of the head end neck cancer]. / Autofluorescencja tkankowa w diagnostyce i ocenie skutecznosci leczenia nowotworów glowy i szyi.

There are still serious diagnostic and therapeutic challenges in case of carcinoma of the larynx, pharynx and oral cavity most often in patients after radiotherapy. An appropriate choice of a site to take a specimen for histopathology is crucial with regard to establish a diagnosis and find an origin of primary lesion and local recurrence. In this regard our attention is focused on those diagnostic methods which enable to detect even small pathologic lesions. Among them is tissue autofluorescence (DAFE). Its advantages are non-invasiveness, high sensitivity and repeatability. The aim of the study was to validate DAFE as a method for early diagnosis and monitoring of treatment efficacy of head and neck cancers. Forty seven patients were included. Each patient had a fibroscopic investigation with the use of white light and light inducing the tissue fluorescence. A specimen for the histopathology was taken from each site which showed fluorescence in order to verify the diagnosis. The presence of neoplastic lesions was always confirmed with histopathology in parallel with the assessment of the lesion area with fluorescence. DAFE is a non-invasive and useful method for the detection of early cancer lesions with the potential to assess the advances of the disease and to monitor its progress.

[Surgery treatment of salivary gland tumors]. / Leczenie chirurgiczne guzów nowotworowych slinianek.

INTRODUCTION: Tumors of parotid gland represent a diverse group of neoplasms with varied clinical behaviors. MATERIAL AND METHODS: The diagnostic methods and treatment of salivary gland tumors are presented in this paper. The authors describe the group of 44 patients with salivary gland tumors treated surgically in ENT Departament Laryngology of Medical University in Lódz. Enucleation, partial and total parotidectomy were performed. Neurosign 100 unit was used during parotidectomy to monitoring course of facial nerve. RESULTS: Tumor mixtus was a dominate type of tumors. Only three patients with malignant tumor had facial nerve paralysis after surgery. CONCLUSIONS: (1) The extent of the surgical procedure depends on histopathologic diagnosis. (2) Using of facial nerve monitoring during parotid surgery improves its outcomes.

[Isolated sarcoidosis of the neck--a report of two cases]. / Izolowana sarkoidoza szyi--opis dwóch przypadków.

Although the first description of sarcoidosis comes from 1877, its etiology still has not been determined. Its frequency in Europe is estimated at 10-80 new cases per 100 thousand people. Sarcoidosis is a granulomatous disease, manifesting mostly in enlargement of lymph nodes of a pulmonary hilus, in changes in pulmonary parenchyma and skin, eyeball, spleen and liver. Changes in about 30% cases also occur in lymph nodes of other regions (neck or retroperitoneal lymph nodes). Changes in salivary glands and heart have also been reported. Isolated sarcoidosis of peripheral lymph nodes occurs rarely. The paper presents two cases of the isolated sarcoidosis of neck lymph nodes. One of them relates to a 70-year-old woman, in which a lymph node sarcoidosis preceded by several months the occurrence of a typical systemic sarcoidosis engaging pulmonary hiluses. The other case is a 30-year-old man, in which the isolated sarcoidosis of lymph nodes was originally diagnosed as a tumor of a submandibular salivary gland. The difficulties in diagnostics of the isolated neck sarcoidosis is discussed as well as its atypical course in both described cases.

[Castleman disease--a case report]. / Choroba Castlemana--opis przypadku.

The Castleman disease was described for the first time in 1956 by Castleman. It is a rare disease of the lymphatic tissue, manifesting in excessive proliferation of lymphocites B and plasmatic cells in lymph nodes. The etiopathogenesis if the disease still is unknown. The paper presents a case of the Castlemana disease (hyaline vascular type) in 20-year-old man with unilateral enlargement of neck lymph nodes. The case exemplifies the difficulties met in diagnostics and treatment of the Castleman disease.

[Diagnostic problems in a case of the pyriform sinus carcinoma in a man]. / Trudnosci diagnostyczne w przypadku raka zachylka gruszkowatego u mezczyzny.

Despite a use of many diagnostic tools to assess the stage of the carcinoma of hypopharynx and larynx various problems can still arise. A 45 years old man was admitted with an initial diagnosis of carcinoma of the hypopharynx with metastases to neck lymphnodes (Tin situ N1). Computed tomography of the neck revealed pathologic remodeling of the thyroid cartilage. An oncologist decided to commence a chemotherapy. After 4 cycles of chemotherapy a second CT scan revealed a suspected neoplastic infiltration of the cricoid and thyroid cartilages. After that the patients was disqualified from both radio- and chemotherapy. The consulting laryngologist did not find any pathologies in the larynx and hypopharynx. On palpation there were no enlarged neck lymph nodes. The second specimen taken from the right pyriform sinus was a loosen fragment of the epithelium with the Ca male differentiatum G3. The positron emission tomography imaging found a suspected site 11 mm in diameter situated in front of the carotid vessels. The neoplastic infiltration of the larynx was not confirmed. The patient started the radiotherapy. We are of the opinion that in the presented case the erroneous interpretation of the CT scan was a likely consequence of the improper setting of a window of brightness and contrast. Strong artifacts are also observed in 3D imaging. Another cause of the diagnostic problems could stem form an unfinished calcification of the cartilages which produced an image of irregular areas of calcification and rarely diagnosed T in situ in a pyriform sinus.