Advanced classical mechanics

This book is designed to serve as a textbook for postgraduates, researchers of applied mathematics, theoretical physics and students of engineering who need a good understanding of classical mechanics. In this book emphasis has been placed on the logical ordering of topics and appropriate formulation of the key mathematical equations with a view to imparting a clear idea of the basic tools of the subject and improving the problem solving skills of the students. The book provides a largely self-contained exposition to the topics with new ideas as a smooth continuation of the preceding ones. It is expected to give a systematic and comprehensive coverage of the methods of classical mechanics

Hippocampal relative expression of <i>Gad65</i>, <i>Gad67</i>, <i>Gfap</i>, <i>Tnf-α</i>, <i>Npy</i> and <i>Ppia</i> genes comparing 0h and 6h of <i>postmortem</i> intervals.

<p>Values are mean ± SEM, n = 6 per group, Unpaired t-test with Welch´s correction, *p< 0.05 and ***p<0.001.</p

Using <i>Postmortem</i> hippocampi tissue can interfere with differential gene expression analysis of the epileptogenic process

<div><p>Neuropathological studies often use autopsy brain tissue as controls to evaluate changes in protein or RNA levels in several diseases. In mesial temporal lobe epilepsy (MTLE), several genes are up or down regulated throughout the epileptogenic and chronic stages of the disease. Given that <i>postmortem</i> changes in several gene transcripts could impact the detection of changes in case-control studies, we evaluated the effect of using autopsy specimens with different <i>postmortem</i> intervals (PMI) on differential gene expression of the Pilocarpine (PILO)induced <i>Status Epilepticus</i> (SE) of MTLE. For this, we selected six genes (<i>Gfap</i>, <i>Ppia</i>, <i>Gad65</i>, <i>Gad67</i>, <i>Npy</i>, and <i>Tnf-α</i>) whose expression patterns in the hippocampus of PILO-injected rats are well known. Initially, we compared hippocampal expression of naïve rats whose hippocampi were harvested immediately after death (0h-PMI) with those harvested at 6h <i>postmortem</i> interval (6h-PMI): <i>Npy</i> and <i>Ppia</i> transcripts increased and <i>Tnf-α</i> transcripts decreased in the 6h-PMI group (p<0.05). We then investigated if these PMI-related changes in gene expression have the potential to adulterate or mask RT-qPCR results obtained with PILO-injected rats euthanized at acute or chronic phases. In the acute group, <i>Npy</i> transcript was significantly higher when compared with 0h-PMI rats, whereas <i>Ppia</i> transcript was lower than 6h-PMI group. When we used epileptic rats (chronic group), the RT-qPCR results showed higher <i>Tnf-α</i> only when compared to 6h-PMI group. In conclusion, our study demonstrates that PMI influences gene transcription and can mask changes in gene transcription seen during epileptogenesis in the PILO-SE model. Thus, to avoid erroneous conclusions, we strongly recommend that researchers account for changes in <i>postmortem</i> gene expression in their experimental design.</p></div

Selection of the most suitable reference genes for 0h and 6h PMI in the hippocampus of Naïve rats.

<p>Expression stability measurements for the 5 reference genes calculated by geNorm (A) and NormFinder (B). The x-axis from left to right indicates the ranking of the genes according to their expression stability; lower values indicate higher expression stability. C) Determination of the optimal number of reference genes for normalization by geNorm. The Software calculates the normalization factor from at least two genes at which the variable V defines the pair-wise variation between two sequential normalization factors.</p

Genetic susceptibility in Juvenile Myoclonic Epilepsy: Systematic review of genetic association studies

<div><p>Background</p><p>Several genetic association investigations have been performed over the last three decades to identify variants underlying Juvenile Myoclonic Epilepsy (JME). Here, we evaluate the accumulating findings and provide an updated perspective of these studies.</p><p>Methodology</p><p>A systematic literature search was conducted using the PubMed, Embase, Scopus, Lilacs, epiGAD, Google Scholar and Sigle up to February 12, 2016. The quality of the included studies was assessed by a score and classified as low and high quality. Beyond outcome measures, information was extracted on the setting for each study, characteristics of population samples and polymorphisms.</p><p>Results</p><p>Fifty studies met eligibility criteria and were used for data extraction. With a single exception, all studies used a candidate gene approach, providing data on 229 polymorphisms in or near 55 different genes. Of variants investigating in independent data sets, only rs2029461 SNP in GRM4, rs3743123 in CX36 and rs3918149 in BRD2 showed a significant association with JME in at least two different background populations. The lack of consistent associations might be due to variations in experimental design and/or limitations of the approach.</p><p>Conclusions</p><p>Thus, despite intense research evidence established, specific genetic variants in JME susceptibility remain inconclusive. We discussed several issues that may compromise the quality of the results, including methodological bias, endophenotype and potential involvement of epigenetic factors.</p><p>PROSPERO registration number</p><p><a href="http://www.crd.york.ac.uk/PROSPERO/display_record.asp?ID=CRD42016036063" target="_blank">CRD42016036063</a></p></div

Flow diagram of study identification.

<p><i>From</i>: MoherD, Libeiati A, TetzlaffJ, Allman DG, The PRISMA Group {2009). <i>Preferred Reporting</i> /terns for Systematic Reviews and <i>Meta- Analyses</i>: The PRISMA Statement. PLoS Med 6(7): e1000097. doi:<a href="https://doi.org/10.1371/journal.pmed1000097" target="_blank">10.1371/journal.pmed1000097</a>. <b>For more information, visit</b> <a href="http://www.prisma-statement.org" target="_blank">www.prisma-statement.org</a>.</p

Identification of microRNAs with Dysregulated Expression in Status Epilepticus Induced Epileptogenesis

<div><p>The involvement of miRNA in mesial temporal lobe epilepsy (MTLE) pathogenesis has increasingly become a focus of epigenetic studies. Despite advances, the number of known miRNAs with a consistent expression response during epileptogenesis is still small. Addressing this situation requires additional miRNA profiling studies coupled to detailed individual expression analyses. Here, we perform a miRNA microarray analysis of the hippocampus of Wistar rats 24 hours after intra-hippocampal pilocarpine-induced Status Epilepticus (H-PILO SE). We identified 73 miRNAs that undergo significant changes, of which 36 were up-regulated and 37 were down-regulated. To validate, we selected 5 of these (10a-5p, 128a-3p, 196b-5p, 352 and 324-3p) for RT-qPCR analysis. Our results confirmed that miR-352 and 196b-5p levels were significantly higher and miR-128a-3p levels were significantly lower in the hippocampus of H-PILO SE rats. We also evaluated whether the 3 miRNAs show a dysregulated hippocampal expression at three time periods (0h, 24h and chronic phase) after systemic pilocarpine-induced status epilepticus (S-PILO SE). We demonstrate that miR-128a-3p transcripts are significantly reduced at all time points compared to the naïve group. Moreover, miR-196b-5p was significantly higher only at 24h post-SE, while miR-352 transcripts were significantly up-regulated after 24h and in chronic phase (epileptic) rats. Finally, when we compared hippocampi of epileptic and non-epileptic humans, we observed that transcript levels of miRNAs show similar trends to the animal models. In summary, we successfully identified two novel dysregulated miRNAs (196b-5p and 352) and confirmed miR-128a-3p downregulation in SE-induced epileptogenesis. Further functional assays are required to understand the role of these miRNAs in MTLE pathogenesis.</p></div

RT-qPCR validation of microarray results.

<p>miRs 196b-5p and 352 were significantly increased and miR-128a-3p was significantly reduced in 24h post-SE hippocampus compared with control group. miRs 10a-5p and 324-3p did not show statistically significant differences. (values are mean±SEM, n = 5-6 in each group, *p < 0.05, Unpaired t test).</p

Relative expression of miR-128a-3p and miR-196b-5p in hippocampi from epileptic and non-epileptic humans.

<p>RT-qPCR analysis (values are meanSEM, biopsy specimens from TLE-HS patients (n = 10-14) and non-epileptic autopsy samples (n = 4), Mann Whitney test).</p

GO Pathway significantly overrepresented among the miR-196b-5p validated targets.

<p>GO Pathway significantly overrepresented among the miR-196b-5p validated targets.</p