Karolina Gołąbek

Association between omentin-1, bone metabolism markers, and cytokines of the RANKL/RANK/OPG system in girls with anorexia nervosa

INTRODUCTION Omentin-1, secreted by visceral adipose tissue, has been indicated in the regulation of bone metabolism in girls with anorexia nervosa (AN). The aim of the study was to evaluate the relationship between omentin-1 and bone metabolism in girls with AN as well as the potential involvement of OPG and RANKL in this relationship. MATERIAL AND METHODS Serum omentin-1, OC, CTx, OPG, and sRANKL were determined by ELISA in 49 girls with AN and in 30 healthy controls, aged 13 to 17 years. RESULTS Girls with AN exhibited significant reduction in body weight, BMI, and Cole index as well as a significant increase in serum omentin-1 levels, compared to healthy participants. These changes were associated with a significant decrease in serum OC and CTx levels and a significant increase in OPG and sRANKL while the OC/CTx and OPG/sRANKL ratios were significantly decreased. BMI and the Cole index correlated negatively and significantly with omentin-1 levels, positively with CTx levels and the OC/CTx ratio in the control group (C), girls with AN, and all study participants (C + AN). Girls with AN showed a significant negative correlation between BMI, the Cole index, and OPG levels. The combined group (C + AN) showed a significant positive correlation between BMI, the Cole index, and the OPG/sRANKL ratio. Omentin-1 levels correlated negatively and significantly with OC and CTx levels as well as with the OC/CTx and OPG/sRANKL ratios in the C, AN, and C + AN groups. CONCLUSIONS The relationship between omentin-1, bone markers, and the OC/CTx and OPG/sRANKL ratios observed in girls with AN indicates the involvement of this adipokine in the regulation of dynamic balance between bone formation and resorption processes. Omentin-1 might exert a negative effect on bone remodelling in girls with AN by inhibiting both bone formation and resorption. The OPG/sRANKL system plays an important role in the latter.

Selected mechanisms of molecular resistance of Candida albicans to azole drugs

A phenomenon of increasing resistance of Candida spp. to azoles has been observed for several years now. One of the mechanisms of lack of sensitivity to azoles is associated with CDR1, CDR2, MRD1 genes (their products are active transport pumps conditioning drug efflux from pathogens cell), and ERG11 gene (encoding lanosterol 14α-demethylase). Test material was 120 strains of Candida albicans (60 resistant and 60 susceptible to azole drugs) obtained from clinical samples. The first stage of experiment assessed the expression of CDR1, CDR2, MDR1 and ERG11 genes by Q-PCR. The impact of ERG11 genes mutations on the expression of this gene was analysed. The final stage of the experiment assessed the level of genome methylation of Candida albicans strains. An increase in the expression of CDR2, MDR1 and ERG11 was observed in azole-resistant strains of Candida albicans in comparison to strains sensitive to this class of drugs. Furthermore, 19 changes in the sequence of ERG11 were detected in tested strains. Four of the discovered mutations: T495A, A530C, G622A and A945C led to the following amino acid substitutions: D116E, K128T, V159I and E266D, respectively. It has also been found that statistically five mutations: T462C, G1309A, C216T, C1257T and A945C affected the expression of ERG11. The applied method of assessing the level of methylation of Candida albicans genome did not confirm its role in the development of resistance to azoles. The results indicate however, that resistance of Candida albicans strains to azole drugs is multifactorial.

Potential use of histone deacetylase inhibitors in cancer therapy

Epigenetics is a branch of science that focuses on mechanisms related to control and modification of expression of genetic material without any changes to its sequences. Such mechanisms include post-translational modifications of histones. It is widely known that carcinogenesis is related to hypoacetylation of genes that influence apoptosis, the cell cycle, cell signaling, the immunologic response, angiogenesis and occurrence of metastasis. Currently conducted research focuses on several strategies related to epigenetic therapy. One such strategy is based on the use of histone deacetylase inhibitors. This paper presents mechanisms through which these compounds work and a summary of their characteristics. It also includes a review of clinical tests related to histone deacetylase inhibitors, as well as their relationship with other chemotherapeutic methods. A better understanding of the involved mechanisms will provide a rational basis to improve the therapeutic outcome of available antitumor agents.

TGF-β1, bone metabolism, osteoprotegerin, and soluble receptor activator of nuclear factor-kB ligand in girls with anorexia nervosa

INTRODUCTION Numerous investigations, and especially in vitro studies, indicate that TGF-β1 may act as an important regulator of bone remodelling. Thus, it could be expected that disturbances of this cytokine production observed by several researchers might play a role in the mechanism leading to the development of osteoporosis in girls with anorexia nervosa (AN). The aim of the study was to determine whether 1) girls with AN exhibited a relationship between TGF-β1 and bone metabolism (as assessed based on serum OC and CTx concentrations) and 2) whether OPG and sRANKL might modify the possible relationship between TGF-β1 and bone metabolism. MATERIAL AND METHODS Serum concentrations of TGF-β, OC, CTx, OPG, and its soluble ligand sRANKL were determined by ELISA in 60 girls with AN and in 20 healthy controls (C). All study participants were aged 13 to 17 years. RESULTS Body weight, BMI, BMI-SDS and the Cole index, serum TGF-β1, OC, CTx, and the OPG/sRANKL ratio were significantly reduced, while OPG and sRANKL levels were significantly increased, in girls with AN compared to healthy participants. BMI and the Cole index correlated negatively and significantly with serum CTx and OPG (AN group) or CTx only (groups C and C + AN). Girls with AN showed a positive and significant correlation between the Cole index and serum TGF-β1. The combination group (C + AN) showed a positive and significant correlation between BMI, the Cole index, and the OPG/sRANKL ratio and TGF-β1 concentration, while TGF-β1 correlated positively and significantly with OC concentrations and the OPG/sRANKL ratio. The Cole index and BMI were identified to be significant and independent predictors of CTx (C, AN, and C+AN groups) and OPG (AN group); the Cole index, BMI, and TGF-β1 independently predicted the OPG/sRANKL ratio (C, AN, and C + AN groups); TGF-β1 was found to be an independent predictor of OC (C + AN group). CONCLUSIONS Changes in bone markers, OPG, and/or OPG/sRANKL ratio observed in girls with AN are associated with changes in serum TGF-β1 concentrations. TGF-β1 suppression in girls with AN might lead to disturbances in the relationship between bone metabolism and the OPG/sRANKL system, which, in turn, might compromise the mechanism compensating for bone remodelling disturbances. (Endokrynol Pol 2016; 67 (5): 493-500).

Vaspin and selected indices of bone status in girls with anorexia nervosa

INTRODUCTION In vitro studies indicate that vaspin may act as a regulator of bone metabolism. The aim of the study was to evaluate the relationship between vaspin and bone metabolism in girls with anorexia nervosa (AN), as well as the potential involvement of OPG and RANKL in this relationship. MATERIAL AND METHODS Serum vaspin, OC, CTx, OPG, and sRANKL were determined by ELISA in 50 girls with AN and in 30 healthy controls aged 13 to 17 years. RESULTS Girls with AN exhibited significant reduction in body weight, BMI, and Cole index as well as a significant increase in serum level of vaspin compared to healthy participants. These changes were associated with a significant decrease in serum OC and CTx levels and a significant increase in OPG and sRANKL, while the OPG/sRANKL ratio was significantly decreased. BMI and Cole index correlated negatively and significantly with CTx levels in the control group (C), girls with AN, and all study participants (C+AN). Girls with AN showed a significant negative correlation between BMI, the Cole index, and OPG levels. The combination group (C+AN) showed a significant positive correlation between BMI, Cole index, and the OPG/sRANKL ratio. In this group of girls vaspin levels correlated positively and significantly with sRANKL and negatively with body weight, BMI, Cole index, and OPG/sRANKL ratio. Girls with AN showed a significant negative correlation between vaspin levels and the OPG/sRANKL ratio. CONCLUSIONS Undernourishment and associated deficit of adipose tissue may result in inadequate vaspin production and bone metabolism disorders in girls with AN. Vaspin acts as a coordinator of the dynamic balance between bone formation and resorption processes; its action is affected by the cytokines of the RANKL/RANK/OPG system. Changes in the relationships between vaspin, bone markers, OPG, and RANKL might contribute to the development of osteoporosis in girls with AN. (Endokrynol Pol 2016; 67 (6): 599-606).

The Content of Reticulocyte Hemoglobin and Serum Concentration of the Soluble Transferrin Receptor for Diagnostics of Anemia in Chronically Hemodialyzed Patients

BACKGROUND Chronic renal disease constitutes a serious worldwide clinical problem. An important issue arising early during the treatment of renal failure is anemia. Patients in the end-stage of renal disease chronically treated with hemodialysis frequently suffer from anemia with iron deficiency. OBJECTIVES The aim of the study was to evaluate the usefulness of determining the reticulocyte hemoglobin content and serum concentration of soluble transferrin receptor in the detection of anemia caused by iron deficiency in comparison with the classic markers of iron circulation in serum in chronic dialysis patients with ESRD. MATERIAL AND METHODS 66 sets of hematologic results and iron turnover rates were analyzed, sampled from hemodialyzed patients (test group), as well as 34 sets of the same results taken from healthy people (control group). Statistically significant variables were found and a stepwise backward discriminant analysis was performed for them. RESULTS The results showed that dialyzed patients have a significantly lower serum concentration of hemoglobin, CHr, HCT, TSAT, Fe and TIBC and significantly higher serum concentration of sTfR, ferritin and C-reactive protein compared to the control group. Based on the results of discriminant analysis, we proposed a scheme for assessing the risk of anemia. CONCLUSIONS The concentrations of hemoglobin, soluble transferrin receptor, iron in the serum and C-reactive protein turned out to be the most useful for diagnostic purposes. Moreover, the concentration of soluble transferrin receptor confirmed its high diagnostic value in the detection of iron deficiency-based anemia in patients undergoing dialysis for chronic renal failure at the end-stage compared to conventional iron turnover ratios in the serum.

Prevalence of Borrelia burgdorferi sensu lato in Ixodes ricinus ticks collected from southern Poland.

In 2008–2011 ticks were collected from southern Poland. Out of 6336 individuals collected and identified as Ixodes ricinus, 768 (2 larvae, 84 nymphs, 417 females, 265 males) were included in molecular study. The aim of this study was to investigate the prevalence and types of genospecies of Borrelia burgdorferi sensu lato in ticks. The polymerase chain reaction (PCR) was applied to detect the presence of pathogens in ticks. Subsequently the amplified DNA was digested with TasI enzyme. The infection rate was 15% (116) of examined ticks. PCR-RFLP analysis allowed distinguishing three genospecies of B. burgdorferi s.l.: B. burgdorferi sensu stricto, B. afzelii, and B. garinii. RFLP analyses of 116 positive samples revealed 96 (83%) monoinfections and 13 (11%) coinfections, whereas unidentified genospecies were present in 7 (6%) of positive samples. In the case of monoinfections, B. burgdorferi s.s. was the predominant species of pathogen in infected ticks - 61.4%. Other genospecies: B. garinii and B. afzelii were detected in 22.9% and 15.6% of the samples, respectively. To sum up, 15 % of ticks were infected by B. burgdorferi s.l which increases the risk of human infections in the recreational areas of southern Poland. Furthermore, there is a need to increase public awareness and implement more preventive measures concerning Lyme disease.

Selected genetic determinants of head and neck squamous cell carcinoma including of some genes for cell cycle control and DNA repair

Raki płaskonabłonkowe narządów głowy i szyi, występujące w różnych strukturach anatomicznych, w tym w jamie ustnej, gardle i krtani, stają się poważnym problemem klinicznym ze względu na wciąż rosnącą liczbę zachorowań, jak i trudności w leczeniu pacjentów. Wśród czynników ryzyka związanych z tą grupą nowotworów dobrze opisano czynniki egzogenne, takie jak: ekspozycja na kancerogeny obecne w dymie tytoniowym, spożywanie alkoholu, niewłaściwe nawyki dietetyczne, nieprawidłowa higiena jamy ustnej, jak i zakażenie wirusem brodawczaka ludzkiego oraz wirusem Epsteina–Barr. Drugą grupą potencjalnych czynników ryzyka są czynniki endogenne, do których należą m.in. uwarunkowania genetyczne i zaburzenia funkcjonowania układu immunologicznego. Od niedawna, ze względu na rozwój technik z zakresu biologii molekularnej, tematem badań stały się polimorfizmy licznych genów kodujących białka o różnych funkcjach. W artykule omówiono problemy związane z podłożem molekularnym rozwoju raka płaskonabłonkowego narządów głowy i szyi. Skupiono się zwłaszcza na polimorfizmach pojedynczego nukleotydu i mutacjach genów uczestniczących w kontroli cyklu komórkowego (TP53, p73, CDKN1A, CDKN2A, MDM2, E2F1, E2F2 i EGFR) oraz w różnych mechanizmach naprawy DNA (XPA, XPB, XPC, XPD, XPF, XPG, ERCC1, OGG1, XRCC1, NBS1, RAD51, BRCA2, XRCC2, XRCC3, XRCC5 i XRCC6). Przedstawione wyniki badań wskazują na niejednorodny obraz uwarunkowań genetycznych nowotworów głowy i szyi. Można jednak zauważyć, że wystąpienie tylko jednego polimorfizmu konkretnego genu w umiarkowanym stopniu wpływało na patogenezę omawianych nowotworów. Dopiero jednoczesne współistnienie kilku polimorfizmów zwiększa ryzyko zachorowania w istotny sposób. Jednak, niezaprzeczalnie należy stwierdzić, że poruszane zagadnienia powinny podlegać dalszym analizom.

Expression profiles of selected genes in tumors and matched surgical margins in oral cavity cancer: Do we have to pay attention to the molecular analysis of the surgical margins?

BACKGROUND Head and neck squamous cell carcinomas (HNSCCs) are associated with an interplay between genetics and the environment; they account for 3% of all diagnosed malignant tumors in men and 2% of those in women. OBJECTIVES The aim of the study was to analyze the significance of TIMP3, SFRP1, SFRP2, CDH1, RASSF1, RORA, and DAPK1 gene expression in head and neck squamous cell carcinoma tumors, and in matching surgical margin samples. We also analyzed the association between clinical parameters and the expression of the selected genes. MATERIAL AND METHODS Following surgical resection, 56 primary HNSCC tumors and matching surgical margin samples were collected from patients at the Clinic of Oncological and Reconstructive Surgery of Maria Skłodowska-Curie Memorial Cancer Center and the Institute of Oncology in Gliwice, Poland. The gene expression levels were analyzed by quantitative reverse transcription (qRT)-PCR. RESULTS SFRP1 gene expression was statistically significantly lower in the tumor samples than in the surgical margins (0.30 ±0.36 vs 0.62 ±0.36; p < 0.01). No correlation was found between gene expression and clinical parameters, except DAPK1, where low expression correlated with alcohol abuse (0.85 ±1.19 vs 1.97 ±3.22; p = 0.074). Moreover, patients with G3 grade tumors, i.e., poorly differentiated tumors, had significantly higher values of DAPK1 gene expression than the G1 (well-differentiated tumors) and G2 (moderately differentiated) groups. CONCLUSIONS There are many different reasons and concepts for altered gene expression in tumors and surgical margin tissue. Tumor heterogeneity and its microenvironment are undoubtedly linked to the biology of HNSCC. In order to understand specific tumor behavior and the microenvironment, further studies are needed. To find markers connected with cancer development and to provide insight into the earliest stages of cancer development, attention should also be focused on molecular analysis of the surgical margins.