Pros and cons of plant nuclear protein enrichment

23rd International PhD Students Conference (MendelNet); NOV 09-10, 2016; Mendel Univ Brno, Fac AgriSciencesNuclear proteome contains important regulatory proteins. To improve the detection of these proteins, Percoll gradient-based fractionation techniques have been developed and optimized. However, owing to the ever increasing sensitivity of identification methods based on liquid chromatography and mass spectrometry, the time and material consuming fractionation methods may no longer be necessary. Here, we show that a Percoll-based nuclear protein fractionation of tomato leaf proteome increased the number of detected proteins, but at least some nuclear proteins were lost or depleted in the process

Effectiveness of coping strategies for victims of cyberbullying

Although cyberbullying is a well-studied online risk, little is known about the effectiveness of various coping strategies for its victims. Therefore, this study on 2,092 Czech children aged 12-18 examined which coping strategies were applied most often and to what extent victims of cyber aggression judged them to be effective. Effectiveness was measured in two ways: the extent to which the coping strategy buffered negative emotions, and the extent to which it stopped the cyber aggression. The subsample of 422 respondents (68% girls) who experienced online attacks was divided into two groups: victims of cyberbullying, who experienced long-term and severe victimization, and victims of less severe online harassment. The selection of strategies was compared between these groups and the most prevalent and effective strategies were identified. Overall, respondents in both groups used more than one strategy at once, but for victims of cyberbullying, most of the strategies proved less effective. Both groups showed similar patterns in their use of technical solutions, and both quite frequently sought social support. Also, both groups showed a low tendency for retaliation. Large differences were found in how respondents employed cognitive responses, such as trying to detach mentally from bullying incidents; these strategies were less frequent among victims of cyberbullying. In terms of effectiveness, victims of harassment considered technical solutions such as blocking contacts on social websites, seeking support, or ignoring the bully on purpose the most effective in stopping victimization, as opposed to victims of cyberbullying, who found them less effective, even though they used them as frequently as victims of online harassment. Differences in the effectiveness of coping strategies are discussed with regard to the intervention and prevention of cyberbullying

Chromosomal damage in peripheral blood lymphocytes of newly diagnosed cancer patients and healthy controls

Background: The majority of human cancers arise from cells unable to maintain genomic stability. Recent prospective studies indicated that enhanced chromosomal aberrations (CAs) frequencies are predictive of gastrointestinal and lung cancers. However, studies on incident cancer patients are lacking; thus, we investigated chromosomal damage in newly diagnosed cancer patients and healthy individuals. Methods: We analyzed chromosomal damage in peripheral blood lymphocytes in a group of 300 incident cancer patients (with different malignancies) in comparison with 300 healthy controls. Results and Conclusions: The frequencies of aberrant cells (ACs) and CAs were significantly higher in patients (2.38 +/- 1.56 and 2.53 +/- 1.69, respectively) as compared with controls (1.81 +/- 1.31 and 1.94 +/- 1.47, respectively, P < 0.01). The percentual difference in chromatid-type aberrations (CTAs) between patients and controls was moderately significant (1.37 +/- 1.20 and 1.11 +/- 0.99, respectively, P < 0.05), whereas the difference in chromosome-type aberrations (CSAs) was stronger (1.16 +/- 1.24 versus 0.83 +/- 1.12, P < 0.01). Using binomial logistic regression, the estimated odds ratios and 95% confidence interval for ACs were 1.33 (1.18-1.49), P < 0.01; for CAs, 1.27 (1.14-1.41), P < 0.01; for CTA 1.24 (1.07-1.44), P < 0.01 and for CSA, 1.27 (1.10-1.47), P < 0.01. By stratifying patients for distinct neoplasia, markers of chromosomal damage were significantly enhanced in patients with breast, prostate and head/neck cancers, whereas no effect was recorded in patients affected by gastrointestinal cancers. The present study shows for the first time evidence of increased chromosomal damage in lymphocytes of incident cancer patients compared with healthy controls. The effects were observed in different cancer types but as the number of patients was relatively small, further studies are warranted

Interferon-α Revisited: Individualized Treatment Management Eased the Selective Pressure of Tyrosine Kinase Inhibitors on BCR-ABL1 Mutations Resulting in a Molecular Response in High-Risk CML Patients.

Bone marrow transplantation or ponatinib treatment are currently recommended strategies for management of patients with chronic myeloid leukemia (CML) harboring the T315I mutation and compound or polyclonal mutations. However, in some individual cases, these treatment scenarios cannot be applied. We used an alternative treatment strategy with interferon-α (IFN-α) given solo, sequentially or together with TKI in a group of 6 cases of high risk CML patients, assuming that the TKI-independent mechanism of action may lead to mutant clone repression. IFN-α based individualized therapy decreases of T315I or compound mutations to undetectable levels as assessed by next-generation deep sequencing, which was associated with a molecular response in 4/6 patients. Based on the observed results from immune profiling, we assumed that the principal mechanism leading to the success of the treatment was the immune activation induced with dasatinib pre-treatment followed by restoration of immunological surveillance after application of IFN-α therapy. Moreover, we showed that sensitive measurement of mutated BCR-ABL1 transcript levels augments the safety of this individualized treatment strategy

Cerebral organoids derived from patients with Alzheimer’s disease with PSEN1/2 mutations have defective tissue patterning and altered development

Summary: During the past two decades, induced pluripotent stem cells (iPSCs) have been widely used to study human neural development and disease. Especially in the field of Alzheimer’s disease (AD), remarkable effort has been put into investigating molecular mechanisms behind this disease. Then, with the advent of 3D neuronal cultures and cerebral organoids (COs), several studies have demonstrated that this model can adequately mimic familial and sporadic AD. Therefore, we created an AD-CO model using iPSCs derived from patients with familial AD forms and explored early events and the progression of AD pathogenesis. Our study demonstrated that COs derived from three AD-iPSC lines with PSEN1(A246E) or PSEN2(N141I) mutations developed the AD-specific markers in vitro, yet they also uncover tissue patterning defects and altered development. These findings are complemented by single-cell sequencing data confirming this observation and uncovering that neurons in AD-COs likely differentiate prematurely

Asymmetric dimethyl arginine induces pulmonary vascular dysfunction via activation of signal transducer and activator of transcription 3 and stabilization of hypoxia-inducible factor 1-alpha

Pulmonary hypertension (PH), associated with imbalance in vasoactive mediators and massive remodeling of pulmonary vasculature, represents a serious health complication. Despite the progress in treatment, PH patients typically have poor prognoses with severely affected quality of life. Asymmetric dimethyl arginine (ADMA), endogenous inhibitor of endothelial nitric oxide synthase (eNOS), also represents one of the critical regulators of pulmonary vascular functions. The present study describes a novel mechanism of ADMA-induced dysfunction in human pulmonary endothelial and smooth muscle cells. The effect of ADMA was compared with well-established model of hypoxia-induced pulmonary vascular dysfunction. It was discovered for the first time that ADMA induced the activation of signal transducer and activator of transcription 3 (STAT3) and stabilization of hypoxia inducible factor la (HIF-1 alpha) in both types of cells, associated with drastic alternations in normal cellular functions (e.g., nitric oxide production, cell proliferation/Ca2+ concentration, production of pro-inflammatory mediators, and expression of eNOS, DDAH1, and ICAM-1). Additionally, ADMA significantly enhanced the hypoxia-mediated increase in the signaling cascades. In summary, increased ADMA may lead to manifestation of PH phenotype in human endothelial and smooth muscle cells via the STAT3/HIF-1 alpha cascade. Therefore this signaling pathway represents the potential pathway for future clinical interventions in PH. (C) 2015 Elsevier Inc. All rights reserved