Expansion of the Parkinson disease-associated SNCA-Rep1 allele upregulates human alpha-synuclein in transgenic mouse brain.

Alpha-synuclein (SNCA) gene has been implicated in the development of rare forms of familial Parkinson disease (PD). Recently, it was shown that an increase in SNCA copy numbers leads to elevated levels of wild-type SNCA-mRNA and protein and is sufficient to cause early-onset, familial PD. A critical question concerning the molecular pathogenesis of PD is what contributory role, if any, is played by the SNCA gene in sporadic PD. The expansion of SNCA-Rep1, an upstream, polymorphic microsatellite of the SNCA gene, is associated with elevated risk for sporadic PD. However, whether SNCA-Rep1 is the causal variant and the underlying mechanism with which its effect is mediated by remained elusive. We report here the effects of three distinct SNCA-Rep1 variants in the brains of 72 mice transgenic for the entire human SNCA locus. Human SNCA-mRNA and protein levels were increased 1.7- and 1.25-fold, respectively, in homozygotes for the expanded, PD risk-conferring allele compared with homozygotes for the shorter, protective allele. When adjusting for the total SNCA-protein concentration (endogenous mouse and transgenic human) expressed in each brain, the expanded risk allele contributed 2.6-fold more to the SNCA steady-state than the shorter allele. Furthermore, targeted deletion of Rep1 resulted in the lowest human SNCA-mRNA and protein concentrations in murine brain. In contrast, the Rep1 effect was not observed in blood lysates from the same mice. These results demonstrate that Rep1 regulates human SNCA expression by enhancing its transcription in the adult nervous system and suggest that homozygosity for the expanded Rep1 allele may mimic locus multiplication, thereby elevating PD risk

Cardiac power output accurately reflects external cardiac work over a wide range of inotropic states in pigs

BACKGROUND: Cardiac power output (CPO), derived from the product of cardiac output and mean aortic pressure, is an important yet underexploited parameter for hemodynamic monitoring of critically ill patients in the intensive-care unit (ICU). The conductance catheter-derived pressure-volume loop area reflects left ventricular stroke work (LV SW). Dividing LV SW by time, a measure of LV SW min- 1 is obtained sharing the same unit as CPO (W). We aimed to validate CPO as a marker of LV SW min- 1 under various inotropic states. METHODS: We retrospectively analysed data obtained from experimental studies of the hemodynamic impact of mild hypothermia and hyperthermia on acute heart failure. Fifty-nine anaesthetized and mechanically ventilated closed-chest Landrace pigs (68 ± 1 kg) were instrumented with Swan-Ganz and LV pressure-volume catheters. Data were obtained at body temperatures of 33.0 °C, 38.0 °C and 40.5 °C; before and after: resuscitation, myocardial infarction, endotoxemia, sevoflurane-induced myocardial depression and beta-adrenergic stimulation. We plotted LVSW min- 1 against CPO by linear regression analysis, as well as against the following classical indices of LV function and work: LV ejection fraction (LV EF), rate-pressure product (RPP), triple product (TP), LV maximum pressure (LVPmax) and maximal rate of rise of LVP (LV dP/dtmax). RESULTS: CPO showed the best correlation with LV SW min- 1 (r2 = 0.89; p < 0.05) while LV EF did not correlate at all (r2 = 0.01; p = 0.259). Further parameters correlated moderately with LV SW min- 1 (LVPmax r2 = 0.47, RPP r2 = 0.67; and TP r2 = 0.54). LV dP/dtmax correlated worst with LV SW min- 1 (r2 = 0.28). CONCLUSION: CPO reflects external cardiac work over a wide range of inotropic states. These data further support the use of CPO to monitor inotropic interventions in the ICU

A napraforgo megporzo rovarai es a hazimeh tevekenysege a napraforgon

Szamos kutató ramutatott arra, hogy a napraforgó viragai izolator alatt feltunoen kisebb aranyban termekenyulnek, ezert a leha szemek aranya izolalas eseten tobbszorosen meghaladja a szabadon elviragzott napraforgó tanyerokon tapasztalható merteket (Anonymus 1950, Radoev 1954, Kaskovszkij 1960, Kirjukin 1960, Kushnir 1960a, 1960b, Sanduleac 1960, Bitkolov 1961, Kurnik es Zelles 1962, 1963, Kamennobrodszkaja 1963, Free es Simpson 1964). A megporzó rovarok fontossagara utalnak azok az eredmenyek is, amelyek bizonyitjak, hogy a hazimeh csaladok kihelyezeset kovetoen nagyobb termest kapunk (Furgała 1954, Cirnu 1960, Hljusztov 1962, Cirnu es Sanduleac 1965), illetve nagyobb hazimeh suruseg eseten (Kurennoj 1957), vagy a meheszethez kozelebb eszrevehetoen nagyobb a termekenyiiles (Kushnir 1960a, 1960b, Kamennobrodszkaja 1953). Free es Simpson (1964) kiserletileg igazoltak, hogy a napraforgó tanyerok viragai masik viragzatról szarmazó pollen hatasara feltunoen nagyobb mertekben termekenyulnek. A megporzó rovarok jelentosege tehat ab ban all, hogy az idegen megporzast elvegzik. Kirjukin (1960) ugyanakkor kimutatta, hogy az eredmenyes termekenyiiles erdek eben a bibe mechanikai,,ingerlese is nelkulozhetetlen. Maskent ugyanis id egen viragpor atvitele ellenere sem ert el jobb kotodest, mint masok ontermekenyiiles eseten

Impact of coverage depth on identification of target sequences using 21-nt simulated reads.

<p>The genome of <i>E</i>. <i>coli</i> Sakai (EC20040078) was used to randomly generate triplicate datasets of simulated 21-nt reads for twelve depths of coverage (36 datasets). The reads from individual datasets were then mapped to the target sequences, and the mean percentage of sequence identity (MPI) was calculated for each dataset. The average and standard deviation of the three MPI values obtained for each dataset at different depths of coverage are shown. An MPI above 90% was used as the threshold for accurate identification.</p

Strains and results of GeneSippr analysis.

<p>^aBased on strain characterization/CHAS results in previous work [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0122928#pone.0122928.ref004" target="_blank">4</a>]</p><p>^bTotal genomic DNA isolated from a single colony</p><p>^cNumber of reads generate (in millions)</p><p>^dEstimated fold coverage of genome achieved with 21-nt reads</p><p>*low coverage was observed for three strains in one run. Analysis was repeated every hour until QC targets were identified. For strains OLC-683, OLC-715, and OLC-684, QC and virulence targets were identified at cycle 125, cycle 175 and cycle 41, respectively.</p><p>Strains and results of GeneSippr analysis.</p

Minimum read length required for accurate identification of target sequences.

<p>The genome of <i>E</i>. <i>coli</i> Sakai (EC20040078) was used to randomly generate triplicate datasets of 0.5M (top left), 1M (top right), 1.5M (bottom left) and 2M simulated reads (bottom right) for twelve read lengths ranging from 18 to 50 nt (144 datasets). The reads from individual datasets were then mapped to the target sequences, and the mean percentage of sequence identity (MPI) was calculated for each dataset. The average and standard deviation of the three mean percentage of sequence identity obtained for each dataset are shown. An MPI above 90% was used as the threshold for accurate identification.</p

GeneSippr: A Rapid Whole-Genome Approach for the Identification and Characterization of Foodborne Pathogens such as Priority Shiga Toxigenic <i>Escherichia coli</i>

<div><p>The timely identification and characterization of foodborne bacteria for risk assessment purposes is a key operation in outbreak investigations. Current methods require several days and/or provide low-resolution characterization. Here we describe a whole-genome-sequencing (WGS) approach (GeneSippr) enabling same-day identification of colony isolates recovered from investigative food samples. The identification of colonies of priority Shiga-toxigenic <i>Escherichia coli</i> (STEC) (i.e., serogroups O26, O45, O103, O111, O121, O145 and O157) served as a proof of concept. Genomic DNA was isolated from single colonies and sequencing was conducted on the Illumina MiSeq instrument with raw data sampling from the instrument following 4.5 hrs of sequencing. Modeling experiments indicated that datasets comprised of 21-nt reads representing approximately 4-fold coverage of the genome were sufficient to avoid significant gaps in sequence data. A novel bioinformatic pipeline was used to identify the presence of specific marker genes based on mapping of the short reads to reference sequence libraries, along with the detection of dispersed conserved genomic markers as a quality control metric to assure the validity of the analysis. STEC virulence markers were correctly identified in all isolates tested, and single colonies were identified within 9 hrs. This method has the potential to produce high-resolution characterization of STEC isolates, and whole-genome sequence data generated following the GeneSippr analysis could be used for isolate identification in place of lengthy biochemical characterization and typing methodologies. Significant advantages of this procedure include ease of adaptation to the detection of any gene marker of interest, as well as to the identification of other foodborne pathogens for which genomic markers have been defined.</p></div