Crystallization of SHARPIN using an automated two-dimensional grid screen for optimization

An N-terminal fragment of human SHARPIN was recombinantly expressed in Escherichia coli, purified and crystallized. Crystals suitable for X-ray diffraction were obtained by a one-step optimization of seed dilution and protein concentration using a two-dimensional grid screen. The crystals belonged to the primitive tetragonal space group P4(3)2(1)2, with unit-cell parameters a = b = 61.55, c = 222.81 Å. Complete data sets were collected from native and selenomethionine-substituted protein crystals at 100 K to 2.6 and 2.0 Å resolution, respectively

Impact of Reddit Discussions on Use or Abandonment of Wearables

Discussion platform, Reddit, is the third most visited website in the US. People can post their questions on this platform to get varying opinions from fellow users, which in turn might also influence their behavior and choices. Wearables are becoming widely adopted, yet challenges persist in their effective long term use because of technical and device related, or personal issues. Therefore, by employing sentiment analysis, this paper aims to analyze how decisions of use or abandonment of wearables are influenced by discussions on Reddit. The results are based on the analysis of 6680 posts and their associated 50,867 comments posted between December 2015 - December 2017 on the subreddit (user created groups) on android wear. Our results show that sentiment of the discussion is majorly dictated by the sentiment of the post itself, and people decide to continue using their devices when fellow Redditors offer them workarounds, or the discussion receives majority of positive or fact-driven neutral comments

Precision cancer monitoring using a novel, fully integrated, microfluidic array partitioning digital PCR platform.

A novel digital PCR (dPCR) platform combining off-the-shelf reagents, a micro-molded plastic microfluidic consumable with a fully integrated single dPCR instrument was developed to address the needs for routine clinical diagnostics. This new platform offers a simplified workflow that enables: rapid time-to-answer; low potential for cross contamination; minimal sample waste; all within a single integrated instrument. Here we showcase the capability of this fully integrated platform to detect and quantify non-small cell lung carcinoma (NSCLC) rare genetic mutants (EGFR T790M) with precision cell-free DNA (cfDNA) standards. Next, we validated the platform with an established chronic myeloid leukemia (CML) fusion gene (BCR-ABL1) assay down to 0.01% mutant allele frequency to highlight the platform's utility for precision cancer monitoring. Thirdly, using a juvenile myelomonocytic leukemia (JMML) patient-specific assay we demonstrate the ability to precisely track an individual cancer patient's response to therapy and show the patient's achievement of complete molecular remission. These three applications highlight the flexibility and utility of this novel fully integrated dPCR platform that has the potential to transform personalized medicine for cancer recurrence monitoring

LUBAC synthesizes linear ubiquitin chains via a thioester intermediate

The linear ubiquitin chain assembly complex (LUBAC) is a RING E3 ligase that regulates immune and inflammatory signalling pathways. Unlike classical RING E3 ligases, LUBAC determines the type of ubiquitin chain being formed, an activity normally associated with the E2 enzyme. We show that the RING-in-between-RING (RBR)-containing region of HOIP—the catalytic subunit of LUBAC—is sufficient to generate linear ubiquitin chains. However, this activity is inhibited by the N-terminal portion of the molecule, an inhibition that is released upon complex formation with HOIL-1L or SHARPIN. Furthermore, we demonstrate that HOIP transfers ubiquitin to the substrate through a thioester intermediate formed by a conserved cysteine in the RING2 domain, supporting the notion that RBR ligases act as RING/HECT hybrids

Identifikacija biološki nerazgradivih zagađivača u riječnoj vodi

Over 100 non-biodegradable organic compounds, accumulated on a carbon filter of a Rhine waterworks, were isolated and identified by means of combined gas chromatography - mass spectrometry. The identified pollutants belong to five different classes: aliphatic as well as aromatic chlorinated hidrocarbons, nitro aromatic compounds, aromatic ethers, tert-butyl substituted phenols and phthalic acid esters. The potential toxicological hazard of the non-biodegradable organic compounds is briefly discussed.Biološki nerazgradivi organski spojevi djeluju nepovoljno na kvalitetu riječne vode koja služi za ljudsku upotrebu i prouzrokuju sekundarne efekte utječući na ekologiju rijeke. Ti se biološki nerazgradivi organski spojevi nakupljaju na ugljenim filtrima vodovoda. Za identifikaciju biološki nerazgradivih organskih spojeva, uzorak ugljenog filtra njemačkog vodovoda s donjeg toka rijeke Rajne, ekstrahiran je s organskim otapalima. Ekstrakti su separirani kromatografskim tehnikama te identificirani pomoću spektrometra masa vezanog uz plinski kromatograf. Neovisno o tome spojevi su identificirani mjerenjem vremena zadržavanja te pomoću infracrvene spektroskopije nakon hvatanja uzorka s maksimuma. Identificirane je više od 100 organskih spojeva. Većina pripada klasama kloriranih alifatskih i aromatskih ugljikovodika, aromatskih nitrospojeva i terc-butil supstituiranih aromatskih ugljikovodika

Molecular insights into RBR E3 ligase ubiquitin transfer mechanisms

National Institute of General Medical Sciences R01 GM0880555T32 GM007270, Francis Crick Institute FCI01, Cancer Research UK, Medical Research Council U117565398, Wellcome Trus

Оценка средней скорости на 10-и метровой глубине для разрезов с высокоскоростным верхним слоем при микрорайонировании

Описываются способы оценки средней скорости поперечной волны на 10-и метровой глубине для разрезов, верхняя часть которых представлена уплотненным насыпным грунтом или мерзлыми породами

Does Subthalamic Deep Brain Stimulation Impact Asymmetry and Dyscoordination of Gait in Parkinson’s Disease?

Background. Subthalamic deep brain stimulation (STN-DBS) is an effective treatment for selected Parkinson’s disease (PD) patients. Gait characteristics are often altered after surgery, but quantitative therapeutic effects are poorly described. Objective. The goal of this study was to systematically investigate modifications in asymmetry and dyscoordination of gait 6 months postoperatively in patients with PD and compare the outcomes with preoperative baseline and to asymptomatic controls without PD. Methods. A convenience sample of thirty-two patients with PD (19 with postural instability and gait disorder (PIGD) type and 13 with tremor dominant disease) and 51 asymptomatic controls participated. Parkinson patients were tested prior to the surgery in both OFF and ON medication states, and 6-months postoperatively in the ON stimulation condition. Movement Disorder Society-Unified Parkinson’s Disease Rating Scale (MDS-UPDRS) I to IV and medication were compared to preoperative conditions. Asymmetry ratios, phase coordination index, and walking speed were assessed. Results. MDS-UPDRS I to IV at 6 months improved significantly, and levodopa equivalent daily dosages significantly decreased. STN-DBS increased step time asymmetry (hedges’ g effect sizes [95% confidence interval] between pre- and post-surgery: .27 [-.13, .73]) and phase coordination index (.29 [-.08, .67]). These effects were higher in the PIGD subgroup than the tremor dominant (step time asymmetry: .38 [-.06, .90] vs .09 [-.83, 1.0] and phase coordination index: .39 [-.04, .84] vs .13 [-.76, .96]). Conclusions. This study provides objective evidence of how STN-DBS increases asymmetry and dyscoordination of gait in patients with PD and suggests motor subtypes‐associated differences in the treatment response