Impact of vitamin D metabolism on clinical epigenetics

The bioactive vitamin D (VD) metabolite, 1,25-dihydroxyvitamin D3 regulates essential pathways of cellular metabolism and differentiation via its nuclear receptor (VDR). Molecular mechanisms which are known to play key roles in aging and cancer are mediated by complex processes involving epigenetic mechanisms contributing to efficiency of VD-activating CYP27A1 and CYP27B1 or inactivating CYP24 enzymes as well as VDR which binds to specific genomic sequences (VD response elements or VDREs). Activity of VDR can be modulated epigenetically by histone acetylation. It co-operates with other nuclear receptors which are influenced by histone acetyl transferases (HATs) as well as several types of histone deacetylases (HDACs). HDAC inhibitors (HDACi) and/or demethylating drugs may contribute to normalization of VD metabolism. Studies link VD signaling through the VDR directly to distinct molecular mechanisms of both HAT activity and the sirtuin class of HDACs (SIRT1) as well as the forkhead transcription factors thus contributing to elucidate complex epigenetic mechanisms for cancer preventive actions of VD

The effect of chair massage on muscular discomfort in cardiac sonographers: a pilot study

<p>Abstract</p> <p>Background</p> <p>Cardiac sonographers frequently have work-related muscular discomfort. We aimed to assess the feasibility of having sonographers receive massages during working hours in an area adjacent to an echocardiography laboratory and to assess relief of discomfort with use of the massages with or without stretching exercises.</p> <p>Methods</p> <p>A group of 45 full-time sonographers was randomly assigned to receive weekly 30-minute massage sessions, massages plus stretching exercises to be performed twice a day, or no intervention. Outcome measures were scores of the <it>Quick</it>DASH instrument and its associated work module at baseline and at 10 weeks of intervention. Data were analyzed with standard descriptive statistics and the separation test for early-phase comparative trials.</p> <p>Results</p> <p>Forty-four participants completed the study: 15 in the control group, 14 in the massage group, and 15 in the massage plus stretches group. Some improvement was seen in work-related discomfort by the <it>Quick</it>DASH scores and work module scores in the 2 intervention groups. The separation test showed separation in favor of the 2 interventions.</p> <p>Conclusion</p> <p>On the basis of the results of this pilot study, larger trials are warranted to evaluate the effect of massages with or without stretching on work-related discomfort in cardiac sonographers.</p> <p>Trial Registration</p> <p>NCT00975026 ClinicalTrials.gov</p

Regulation of hTERT by BCR-ABL at multiple levels in K562 cells

<p>Abstract</p> <p>Background</p> <p>The cytogenetic characteristic of Chronic Myeloid Leukemia (CML) is the formation of the Philadelphia chromosome gene product, BCR-ABL. Given that BCR-ABL is the specific target of Gleevec in CML treatment, we investigated the regulation of the catalytic component of telomerase, hTERT, by BCR-ABL at multiple levels in K562 cells.</p> <p>Methods</p> <p>Molecular techniques such as over expression, knockdown, real-time PCR, immunoprecipitation, western blotting, reporter assay, confocal microscopy, telomerase assays and microarray were used to suggest that hTERT expression and activity is modulated by BCR-ABL at multiple levels.</p> <p>Results</p> <p>Our results suggest that BCR-ABL plays an important role in regulating hTERT in K562 (BCR-ABL positive human leukemia) cells. When Gleevec inhibited the tyrosine kinase activity of BCR-ABL, phosphorylation of hTERT was downregulated, therefore suggesting a positive correlation between BCR-ABL and hTERT. Gleevec treatment inhibited <it>hTERT </it>at mRNA level and significantly reduced telomerase activity (TA) in K562 cells, but not in HL60 or Jurkat cells (BCR-ABL negative cells). We also demonstrated that the transcription factor STAT5a plays a critical role in <it>hTERT </it>gene regulation in K562 cells. Knockdown of STAT5a, but not STAT5b, resulted in a marked downregulation of <it>hTERT </it>mRNA level, TA and hTERT protein level in K562 cells. Furthermore, translocation of hTERT from nucleoli to nucleoplasm was observed in K562 cells induced by Gleevec.</p> <p>Conclusions</p> <p>Our data reveal that BCR-ABL can regulate TA at multiple levels, including transcription, post-translational level, and proper localization. Thus, suppression of cell growth and induction of apoptosis by Gleevec treatment may be partially due to TA inhibition. Additionally, we have identified STAT5a as critical mediator of the <it>hTERT </it>gene expression in BCR-ABL positive CML cells, suggesting that targeting STAT5a may be a promising therapeutic strategy for BCR-ABL positive CML patients.</p

Effects of imatinib and nilotinib on the whole transcriptome of cultured murine osteoblasts.

Numerous clinical observations have confirmed that breakpoint cluster region-abelson fusion oncoprotein tyrosine kinase inhibitors used in leukemia treatment alter bone physiology in a complex manner. The aim of the present study was to analyze the whole transcriptome of cultured murine osteoblasts and determine the changes following treatment with imatinib and nilotinib using Sequencing by Oligonucleotide Ligation and Detection next generation RNA sequencing. This study also aimed to identify candidate signaling pathways and network regulators by multivariate Ingenuity Pathway Analysis. Based on the right-tailed Fisher's exact test, significantly altered pathways including upstream regulators were defined for each drug. The correlation between these pathways and bone metabolism was also examined. The preliminary results suggest the two drugs have different mechanisms of action on osteoblasts, and imatinib was shown to have a greater effect on gene expression. Data also indicated the potential role of a number of genes and signaling cascades that may contribute to identifying novel targets for the treatment of metabolic bone diseases

The Molecular Binding of Resveratrol to Human Serum Albumin using Affinity Capillary Electrophoresis

Resveratrol is a type of natural phenol which is found in the skin of grapes, blueberries and raspberries which is gaining popularity in the health field as it is believed to have anti-cancer, anti-inflammatory, cardio-protective and chemo-preventative effects. Due to the low aqueous solubility of resveratrol, a delivery protein must be used to transport resveratrol from the plasma to the cell-active site. It is hypothesized that human serum albumin (HSA) is the major protein which allows for the transfer of resveratrol into the blood plasma; therefore, it would be essential to perform binding studies between resveratrol and HSA. In this work, an affinity capillary electrophoresis method was developed to determine the binding constant which correlates to the strength of the binding between resveratrol and HSA. In addition, a thermodynamic study was also performed to determine the type of non-covalent binding between the two molecules

Radiostereometric Analysis Permits in Vivo Measurement of Very Small Levels of Wear in TKA

© 2019 by the Association of Bone and Joint Surgeons. Unauthorized reproduction of this article is prohibited. Background Revision of TKA as a result of polyethylene wear is decreasing, but long-term wear performance of polyethylene is still a topic of interest to surgeons and device manufacturers seeking to improve longevity. Measuring wear of modern, wear-resistant implants has been described using radiostereometric analysis (RSA). Performing in vivo measurements would establish whether implant retrieval studies are representative of wear in well-performing knees.Questions/purposesFor a single knee implant system, we sought to determine (1) the linear wear rate using RSA; (2) the association between demographic factors and wear rate; and (3) the association between limb alignment and wear rate.MethodsA total of 49 patients with a minimum followup of 10 years (median, 12 years; range, 10-20 years) were retrospectively selected. During the examined period, 4082 TKAs were performed of which 2085 were the implant examined in this study. There were 71 of these patients who met the criteria including an available full-leg radiograph postoperatively, and 34 of these patients returned for examination along with 15 additional from a separate RSA study that also met the criteria. All patients received a posterior-stabilized, cobalt-chromium-on-conventional polyethylene total knee implant from a single implant system, which was the most commonly used at our institution at the time. Patients underwent standing RSA examinations from 0° to 120° of flexion at a single time point without the use of marker beads. Linear wear rates (including creep) were measured based on intersections between the femoral component and tibial insert models. Associations between wear and patient age at surgery, sex, height, weight, body mass index, tibial insert size, and limb alignment were examined.ResultsUsing the maximum linear wear rate from any flexion angle, the lateral rate was 0.047 mm/year (interquartile range [IQR], 0.034-0.066 mm/year) and the medial rate was 0.052 mm/year (IQR, 0.040-0.069 mm/year). Using the median of the linear wear rates across all flexion angles, the lateral rate was 0.027 mm/year (IQR, 0.017-0.046 mm/year) and the medial rate was 0.038 mm/year (IQR, 0.022-0.054 mm/year). This rate for males was 0.049 mm/year medially (IQR, 0.042-0.077 mm/year) and 0.032 mm/year laterally (IQR, 0.026-0.059 mm/year), and for females was 0.027 mm/year medially (0.016-0.039 mm/year) and 0.020 mm/year laterally (IQR, 0.013-0.032 mm/year). The wear rate for males was greater medially (difference = 0.022 mm/year, p \u3c 0.001) and laterally (difference = 0.012 mm/year, p = 0.008). There were associations between greater wear and increasing height (ρ = 0.48, p \u3c 0.001 medially and ρ = 0.30, p = 0.04 laterally), decreasing body mass index (ρ = -0.31, p = 0.03 medially), and greater implant size (ρ = 0.34, p = 0.02 medially). Increasingly varus leg alignment was associated with greater medial wear (ρ = 0.33, p = 0.02).ConclusionsGreater wear rates were associated with demographic factors and leg alignment. Further RSA wear studies of other modern implant systems would provide complementary information to retrieval studies and valuable data on wear resistance.Clinical RelevanceGood wear resistance was demonstrated by well-performing implants in patients at long-term followup with wear magnitudes in agreement with reported values from retrieval studies