Seminal plasma N-glycome as a new biomarker of environmental exposure associated with semen quality.

Male infertility, a condition that has during the last decade raised significant concern, is a diagnostically demanding and socially sensitive topic. The number of unsolved issues on infertility etiology, especially potential environmental causes, in couples demonstrates the need for further investigations into infertility biomarkers. Semen parameters are often insufficient for reliable profiling of male infertility. Thus, this study aims to evaluate for the first time seminal plasma N-glycosylation as a biomarker of environmental exposure in semen samples from 82 normozoospermic men and 84 men with abnormal semen parameters and compare it with genome damage measured by DNA fragmentation. We obtained information about chronic exposure to environmental factors from the self-reported questionnaire, and determined sperm DNA fragmentation by sperm chromatin dispersion, while N-glycans were characterized with liquid chromatography-mass spectrometry (LC-MS). Based on previously published results, ten N-glycans were selected. Results show that the selected seminal plasma N-glycans were significantly associated with smoking, exposure to pesticides, air pollution, agents emitted during photocopying, alcohol consumption, and obesity. Some N-glycans showed a simultaneous association with DNA fragmentation, semen parameters, and environmental stressors. These subgroups of N-glycans are new potential candidates for biomonitoring of exposure to different environmental factors in men with semen abnormalities.

Inkjet printed highly porous TiO2 films for improved electrical properties of photoanode.

The aim of presented work is to show the improvements obtained in the properties of TiO2 films for dye sensitized solar cells fabricated by inkjet printing using an innovative methodology. We describe the development and properties of TiO2-based inks used in a lab-scale printer, testing various commercial TiO2 pastes. The porosity of the deposited inkjet printed TiO2 films is much higher than using the conventional "doctor blade" deposition technique, as the ink solvent evaporates during the droplet fly from the nozzle to the substrate due to its picoliter volume and the applied heating of a printing stage (70°C). Thanks to higher surface area, the dye sensitized solar cells incorporating inkjet printed TiO2 film gave higher efficiencies (ηmax≈3.06%) than the more compact films obtained by the "doctor blade" method (ηmax≈2.56%). Furthermore, electrochemical analysis indicates that for whole tested thickness range, the inkjet printed layers have higher effective electron diffusion length indicating their better transport properties.

Use of eggs derived from the interspecific charr hybrids to induce androgenetic development of the brook charr (Salvelinus fontinalis Mitchill 1814).

Although, brook charr (Salvelinus fontinalis Mitchill 1814) and Arctic charr (Salvelinus alpinus Linnaeus 1758) are able to cross and give fertile offspring, their androgenetic nucleocytoplasmic hybrids are not viable. To overcome incompatibility between the egg cytoplasm of one charr species and the sperm nucleus of another charr species, application of F1 interspecific hybrids as egg donors for the purpose of androgenesis has been proposed. Here, androgenetic development of the brook charr was successfully induced in the brook charr eggs and the eggs derived from the reciprocal brook charr × Arctic charr F1 hybrids. A working androgenesis protocol included inactivation of the maternal nuclear DNA achieved by irradiation of the eggs with 420 Gy of X-rays, insemination of such treated eggs with the haploid sperm cells and exposition of the haploid androgenetic zygotes to the high hydrostatic pressure shock (51.711 MPa for 4 min) applied 420 min after insemination. Androgenetic larvae that hatched from the brook charr and the hybrid eggs were shown to be homozygous brook charr individuals. Androgenetic individuals exhibited 84 chromosomes and 100 chromosome arms (FN), values characteristic for the brook charr diploid cells. Strategy hybridize first than induce androgenesis should be tested in order to provide androgenetic offspring in other salmonids that are able to cross and produce fertile offspring.

Role of trimethylboron to silane ratio on the properties of p-type nanocrystalline silicon thin film deposited by radio frequency plasma enhanced chemical vapour deposition.

Trimethylboron (TMB) has been receiving attention as a valid alternative to diborane and methane mixtures for the deposition of p-type silicon films for applications in optoelectronic devices such as solar cells. In this paper we report on p-type hydrogenated nanocrystalline silicon carbide (nc-Si:C:H) films produced by standard 13.56 MHz plasma enhanced chemical vapour deposition technique, using TMB as gas source, under high hydrogen dilution (98%) and using high deposition pressures (3 Torr). The films obtained were characterized by spectroscopic ellipsometry (SE), Raman spectroscopy (RS), and electrical measurements to determine their optical, structural and electrical properties. We achieved conductivities as high as 8.3 (omega cm)(-1), one of the highest values of conductivity published to date using TMB with standard rf-PECVD. Spectroscopic ellipsometry modeling revealed that the films growth mechanism proceeds through a sub-surface layer mechanism that leads to the formation of nanocrystalline silicon.

Abnormalities of the P53, MDM2, BCL2 and BAX genes in acute leukemias.

Abnormalities of the P53 network have been implicated in the pathogenesis of acute lymphoblastic leukemia (ALL) and acute myeloblastic leukemia (AML). The purpose of this study was to define P53 gene mutations, to detect MDM2 gene amplification and to estimate mRNA expression of P53, MDM2, BCL2 and BAX genes in patients with ALL and AML. Twenty-five patients with ALL and 65 patients with AML, both recently diagnosed, were included into this study. Exons 5-8 of the P53 gene with flanking intronic sequence were amplified by the polymerase chain reaction (PCR) method and subjected to mutation screening by single-strand conformation polymorphism analysis (SSCP). Mutation of the P53 gene was found in one patient of the 25 with ALL and in five patients of the 65 with AML. Sequence analysis was subsequently performed. One mutation in intronic sequence in ALL and four missense mutations and one silent nucleotide substitution in AML were identified. Amplification of MDM2 gene was detected by multiplex-PCR analysis in only one sample from patient with ALL, but was not observed in any case of AML. To gain further insight into the role of P53 network in the evolution of acute leukemias, the P53, MDM2, BCL2 and BAX mRNAexpressions in portion samples from patients with ALL and AML were analyzed using multiplex RT-PCR. Although a low frequency of molecular disturbances of the P53 and the MDM2 genes was detected in this study, there was a high percentage of cases with increased mRNA level of P53 and MDM2. A high frequency of BCL2 mRNA overexpression and a relatively low frequency of BAX mRNA overexpression detected in both analyzed leukemias in this study, indicate that altered transcription of these genes may be involved in leukemogenesis.

Detection of lymph node metastases of papillary thyroid cancer-comparison of the results of histopathology, immunohistochemistry and reverse transcription-polymerase chain reaction-a preliminary report.

BACKGROUND: The range of lymphadenectomy in differentiated thyroid cancer remains still a matter of controversy because of the lack of reliable diagnostic methods for nodal metastases, other than histopathology. AIM: To compare the results of detection of lymph node metastases of papillary thyroid cancer by conventional histopathology and immunohistochemistry with the results of reverse transcription-polymerase chain reaction for thyroglobulin mRNA. PATIENTS AND METHODS: Each of 166 cervical lymph nodes obtained from 21 patients was divided into two halves: one was used for conventional histopathology and immunohistochemistry, the other part was investigated by molecular examination. RESULTS: We obtained different results from examination of the lymph nodes in six (28.6%) patients. In four patients (19.1%) reverse transcription-polymerase chain reaction (RT-PCR) was more sensitive in detection of positive lymph nodes; in two patients (9.5%) it revealed fewer metastasised lymph nodes than did histopathology. The rest of the patients did not have any differences: 12 (57.1%) of them had negative lymph nodes and three (14.3%) had positive lymph nodes in all examinations. CONCLUSIONS: (1) Thyroglobulin (Tg) RT-PCR is an appropriate method of detection for thyroid cancer cells. (2) In combination with histopathology, it might help to qualify patients' nodal status better.

[Involvement of WT1 gene expression in regulation of P53 and MDM2 proteins function in acute lymphoblastic leukemia]. / Rola ekspresji genu WT1 w regulacji funkcji bialek P53 i MDM2 w ostrej bialaczce limfoblastycznej.

Using sensitive techniques such as reverse transcription polymerase chain reaction (RT-PCR) the expression of WT1 gene in acute lymphoblastic leukemias (ALL) is indicated. High level of mRNA WT1 was only observed in ALL cases with leukemic cells characterized by P53- and MDM2-positive staining in cytometric analysis. The overexpression of P53 protein has not been induced by P53 gene abnormalities and MDM2 protein synthesis was independent from respective gene amplification. The data suggest that WT1 may play a distinct role in the pathophysiology of acute leukemias. It can regulate the function of the main oncoprotein network factors--P53 and MDM2 proteins. There was concluded that the most important mechanism of tissue P53-immunopositivity was connected with the P53 interactions with other oncoproteins, especially with MDM2 and WT1. They have caused different effects in particular cases and several phenotypes of leukemic cells were described. However, the negative tissue staining with anti-P53 monoclonal antibodies can not be evidence of the proper P53 protein function. The immunohistochemical estimations of the P53 level in the cells are insufficient for diagnostic and clinical evaluations. Molecular analyses of P53 and MDM2 genes, as well the WT1 gene transcription, are necessary for the proper characterisation of functional and structural status of P53.