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Various types of neuromuscular diseases differ in symptoms, pathology, and clinical picture but one of their common elements is muscle weakness, which could lead to human motor activities impairment and in many cases to shortening of life span and even death due to respiratory failure. That is why it is very important to better understand the underlying causes of these diseases to be able to implement new methods of treatment more effectively. This paper presents the results of the elemental analysis of human muscular tissues affected by dystrophy and myopathy. For this purpose, the particle-induced X-ray emission method was used, which is perfectly suited for measuring light elements. The samples were analysed for differences in the elemental composition of Na, Mg, P, S, Cl, K, Fe, Zn, and Br. The results were presented in the form of elemental concentration maps and a thorough statistical analysis of the obtained data using the advanced statistical methods.

Introduction: Prothymosin alpha (PTMA), a nuclear oncoprotein involved in cell cycle regulation, is used as a prognostic marker in many cancers. The histopathology of pituitary carcinomas and locally invasive adenomas is indistinguishable from that of benign tumors. A new marker is needed to differentiate these lesions. We evaluated PTMA in pituitary adenomas to determine its usefulness as a prognostic factor of tumor proliferation.Material/Methods: We conducted a retrospective analysis of a group of 27 patients, including 15 females (56%) and 12 males (44%) with a mean age of 58.6±12 years, who underwent pituitary tumor surgery between 2003 and 2012. The Ki-67 and PTMA-nuclear (PTMA-n) and PTMA-cytoplasmic (PTMA-c) indices were determined by immunohistochemical staining. We studied histopathological features, clinical symptoms, and magnetic resonance imaging or computed tomography performed before surgery and one year following surgery to evaluate tumor size and progression.Results: The expression of Ki-67 was revealed in 77.8% of adenomas, PTMA-n in 81.5% and PTMA-c in 92.6%. The mean value of the Ki-67 index was 1.8%, PTMA-n was 1.84%, and PTMA-c was 35.6%. There was a significant positive correlation between Ki-67 and PTMA-n (p=0.009). We did not find any correlation between Ki-67, PTMA-c, and tumor progression. PTMA-n was found to be correlated with tumor size (p=0.045) and was higher in the case of gonadotropinomas (p=0.026).Conclusions: The positive nuclear expression of Ki-67 and PTMA was observed in the majority of pituitary adenomas. Neither the expression of Ki-67 nor that of PTMA-c was related to tumor recurrence or local invasion.

Neoplastic and healthy brain tissues were analysed to discern the changes in the spatial distribution and overall concentration of elements using micro X-ray fluorescence spectroscopy. High-resolution distribution maps of minor and trace elements such as P, S, Cl, K, Ca, Fe, Cu and Zn made it possible to distinguish between homogeneous cancerous tissue and areas where some structures could be identified, such as blood vessels and calcifications. Concentrations of the elements in the selected homogeneous areas of brain tissue were compared between tumours with various malignancy grades and with the controls. The study showed a decrease in the average concentration of Fe, P, S and Ca in tissues with high grades of malignancy as compared to the control group, whereas the concentration of Zn in these tissues was increased. The changes in the concentration were found to be correlated with the tumour malignancy grade. The efficacy of micro X-ray fluorescence spectroscopy to distinguish between various types of cancer based on the concentrations of studied elements was confirmed by multivariate discriminant analysis. Our analysis showed that the most important elements for tissue classification are Cu, K, Fe, Ca, and Zn. This method made it possible to correctly classify histopathological types in 99.93% of the cases used to build the model and in as much as 99.16% of new cases.

The process of carcinogenesis may influence normal biochemical reactions leading to alterations in the elemental composition of the tissue. Total reflection X-ray fluorescence analysis (TXRF) was applied to the elemental analysis of different brain tumors. The following elements were present in all the neoplastic tissues analyzed: K, Ca, Fe, Cu, Zn and Rb. The results of the analysis showed that the elemental composition of a relatively small fragment of tissue represents satisfactorily the biochemical “signature” of a cancer. On the basis of the element concentrations determined, it was possible to differentiate between some types of brain tumors.

ficiency of chemotherapy of malignant gliomas may be combined administration of alkylating agents and those targeted at epithelial growth factor receptor (EGFR). The patients eligible for such therapy should show hypermethylation of MGMT (O6-methylguanineDNA-methyltransferase) promoter as well as amplification of EGFR. Aim of the study: Preliminary assessment of frequency of MGMT hypermethylation and amplification of EGFR co-occurrence in malignant gliomas in patients diagnosed and operated on in the Neurosurgery Department, JUMC in Cracow. Material and methods: 21 consecutive cases of glioblastoma multiforme (W 65%, M 35%, age average 51 y). The methylation status of the MGMT promoter was determined by methylationspecific PCR (MS-PCR). Before MS-PCR, genomic DNA was treated with sodium bisulphite, purified, denatured, precipitated and eluted. EGFR amplification was investigated using the FISH method. Results: With regards to MGMT methylation status and the occurrence of EGFR amplification, four groups of tumours (patients) were established: I – hypermethylation and amplification (4/21, 19.04%); II – hypermethylation, no amplification (6/21, 28.6%); III – no hypermethylation with amplification (4/21, 19.04%); IV – no hypermethylation, no amplification (7/21, 33.3%). Conclusions: Theoretically almost one fifth (19%) of patients with glioblastoma multiforme, i.e. those featuring MGMT hypermethylation and EGFR amplification, could potentially benefit from bimodal chemotherapy with an alkylating agent and an EGFR blocker. Our results confirm other reports indicating that there are no relations between MGMT methylation and EGFR amplification. Financial support by Grant No. K/ZDS/ 001044 Jagiellonian Univ.

Human brain aging is considered to be the leading risk factor for a variety of neurodegenerative alterations. In particular, it is thought that the human substantia nigra might play a pivotal role in age-associated dopamine depletion which could be responsible for neuronal demise and subsequent emergence of different neurological alterations. A plethora of neurochemical redox- and non-redox-driven mechanisms is mainly associated with modifications in the elemental composition of both neuromelanin-pigmented neurons and extraneuronal spaces in the human substantia nigra pars compacta (SNpc). An age-associated variation in the content of Fe, Cu, Zn and Ca has recently received great interest in neurology, as these elements are implicated in different biochemical mechanisms underlying malicious neurodegenerative disorders, including Parkinson's and Alzheimer's diseases. Interestingly, to the best our knowledge, there is lack of a comprehensive study on age-associated variation in the elemental composition of the human SNpc. In that respect, the aim of the present study was to make a preliminary attempt to unravel some of the age-associated mechanisms responsible for the metabolism of some redox-active and redox-inactive elements in the elderly. To do so, substantia nigra tissue specimens, drawn from 37 individuals who deceased without any signs of neurodegeneration, were subjected to spectroscopic studies using synchrotron radiation based X-ray fluorescence. Both neuromelanin-pigmented neurons and extraneuronal areas were studied. It appears that in the neurons, Fe tends to decrease, whilst Cu, Zn and Ca were found to accumulate as an individual gets older.

There is growing evidence that a variety of biochemical processes that underlie the most frequent neurodegenerative diseases may have much in common with those connected with natural aging. It was shown that they involve, among others, lipid peroxidation and/or generation of insoluble in water protein deposits (i.e. alpha-synuclein and/or beta amyloid). Therefore, it is likely that the analysis of changes in both lipid and protein composition may be interesting in the light of any potential pathologies occurring within the dopaminergic system during physiological aging. Thereby, this paper presents a methodology for the analysis of age-related changes in a lipid and protein composition within human subtantia nigra tissue by means of Fourier transform infrared microspectroscopy (FTIRM). Particularly, the changes in the lipid saturation, unsaturation as well as in the protein secondary structure were examined. The studies were carried out on samples from 35 individuals who died without any signs of neurologic dysfunctions. Our results show that the level of lipid saturation increases inside the subtantia nigra tissue with age, though the total content of lipid decreases with age of individuals. Moreover, the statistically significant decrease in the protein content within neuron bodies was observed. Interestingly, it is presented that the content of the anti-parallel beta sheets for neuron bodies decreases from seventh to eighth decades of life and subsequently markedly increases from eighth to ninth decades of life, whilst, as regards extraneuronal spaces, the opposite trends are reported i.e. increase from the seventh to eighth decades, and subsequent decrease in the ninth decade of life. These observations, though preliminary, shed the light on a putative contribution of various pathological lipid- and protein-related processes underlying senescence, suggesting a "biochemical link" between the aetiology of the most common neurodegenerative diseases and physiological aging.

The aims of this paper were to present a reliable morphometric procedure for glioma analysis for preliminary prognosis and to develop a semi-automatic procedure that is easy to use. The data presented are important to the extent that they verify the reliability of the results by showing that they are consistent with the findings from more complicated automatic analytical tools. The objects for analysis were digital images of haematoxylin-eosin stained glioma samples. The overall analysis consisted of digital image analysis and the determination of morphometric parameters. Interestingly, an increase in the mean values of aspect ratio with increasing malignancy grade was found. Moreover, the morphometric parameters in relation to the histological origin of gliomas were examined and it was found that, the cellular nuclei of glioblastoma multiforme reveal the biggest mean values of aspect ratio compared with other gliomas.

Despite the enormous advances in medicine, brain tumours are still among the lesser-known types of tumours and carry the worst prognoses. Transition metals are believed to play an essential role in carcinogenesis. The aim of this study was to determine differences in the average oxidation state and trends in the changes in the chemical environment of iron and zinc contained in healthy and neoplastic tissues of the human brain. For this purpose, X-ray Absorption Spectroscopy was used, which enables the study of disordered matter. The samples were taken intraoperatively and then immediately frozen to slow down chemical processes. Sixteen tumour samples with various malignancy grades were studied as well as one control sample. For each sample four to eight spectra were recorded, with a shift between them not greater than 0.2 eV. In all of the samples, iron occurred in compounds with both Fe(2+) and Fe(3+). However, the ratio of Fe(ii) to Fe(iii) content in the tissue visibly increased with the tumour malignancy grade. The change in the oxidation state of iron did not correlate with the hypoxia level of the tissues. Analysis of EXAFS spectra of zinc atoms showed that the chemical environment of zinc atoms differed with the tumour malignancy grade. Additionally, cryogenic conditions were found to produce positive results in studies of biological samples, whose form under such conditions is close to their native state, without preparation-caused artefacts.

Patients with a PROP1 inactivating mutation present combined pituitary hormone deficiency (CPHD) and pituitary maldevelopment. A retrospective analysis of 31 CPHD patients with a PROP1 mutation revealed two individuals, aged 18 and 4.5 years, who had undergone subtotal surgery to remove pituitary tumors, 16.8 x 12 mm and 9 x 10 x 12 mm in size. Histological reassessment of tissue samples revealed epithelial cells, partially oxyphilic, forming gland-like microcystic structures, most of them filled with eosinophilic colloid. These structures were directly linked with fragments of the posterior lobe. Neither atypia nor any traces of proliferation activity (Ki-67 LI=0%) were noted. Immunohistochemistry showed the presence of all hormonal phenotypes of cells. These findings corresponded to the intermediate lobe of the pituitary gland. For this type of pathology we propose the term 'cystic hyperplasia of the intermediate pituitary lobe' and suggest PROP1 gene assessment in patients with CPHD in order to avoid unnecessary neurosurgical interventions.