Family-wide analysis of poly(ADP-ribose) polymerase activity

The poly(adenosine diphosphate (ADP)-ribose) polymerase (PARP) protein family generates ​ADP-ribose (​ADPr) modifications onto target proteins using ​NAD[superscript +] as substrate. Based on the composition of three ​NAD[superscript +] coordinating amino acids, the H-Y-E motif, each PARP is predicted to generate either poly(ADPr) (PAR) or mono(ADPr) (MAR). However, the reaction product of each PARP has not been clearly defined, and is an important priority since PAR and MAR function via distinct mechanisms. Here we show that the majority of PARPs generate MAR, not PAR, and demonstrate that the H-Y-E motif is not the sole indicator of PARP activity. We identify automodification sites on seven PARPs, and demonstrate that MAR and PAR generating PARPs modify similar amino acids, suggesting that the sequence and structural constraints limiting PARPs to MAR synthesis do not limit their ability to modify canonical amino-acid targets. In addition, we identify ​cysteine as a novel amino-acid target for ADP-ribosylation on PARPs.Rita Allen FoundationSidney Kimmel FoundationNational Cancer Institute (U.S.) (Cancer Center Support (Core) Grant P30-CA14051)National Institutes of Health (U.S.) (Grant RO1GM087465)Kathy and Curt Marble Cancer Research FundWellcome Trust (London, England)European Research Counci

Predicting lymphoma in Sjögren's syndrome and the pathogenetic role of parotid microenvironment through precise parotid swelling recording

Objective: Parotid swelling (PSW) is a major predictor of non-Hodgkin lymphoma (NHL) in primary Sjögren's syndrome (pSS). However, since detailed information on the time of onset and duration of PSW is scarce, this was investigated to verify whether it may lead to further improved prediction. NHL localisation was concomitantly studied to evaluate the role of the parotid gland microenvironment in pSS-related lymphomagenesis. Methods: A multicentre study was conducted among patients with pSS who developed B cell NHL during follow-up and matched controls that did not develop NHL. The study focused on the history of salivary gland and lachrymal gland swelling, evaluated in detail at different times and for different durations, and on the localisation of NHL at onset. Results: PSW was significantly more frequent among the cases: at the time of first referred pSS symptoms before diagnosis, at diagnosis, and from pSS diagnosis to NHL. The duration of PSW was evaluated starting from pSS diagnosis, and the NHL risk increased from PSW of 2-12 months to > 12 months. NHL was prevalently localised in the parotid glands of the cases. Conclusion: A more precise clinical recording of PSW can improve lymphoma prediction in pSS. PSW as a very early symptom is a predictor, and a longer duration of PSW is associated with a higher risk of NHL. Since lymphoma usually localises in the parotid glands, and not in the other salivary or lachrymal glands, the parotid microenvironment appears to be involved in the whole history of pSS and related lymphomagenesis

Overview of fast on-board integrated battery chargers for electric vehicles based on multiphase machines and power electronics

The study provides an extensive overview of on-board integrated chargers for electric vehicles that are based on multiphase (more than three phases) machines and power electronics. A common attribute of all discussed topologies is that they do not require a charger as a separate device since its role is transferred to the already existing drivetrain elements, predominantly a multiphase machine and an inverter. The study demonstrates how additional degrees of freedom that exist in multiphase systems can be conveniently utilised to achieve torque-free charging operation. Therefore, although three-phase (or multiphase) currents flow through machines' stator windings, they do not generate any torque; thus the machines do not have to be mechanically locked. Cost and weight saving is achieved in this way, while the available space is increased. For each topology operating principles are explained, and its control elaborated in detail for both charging and vehicle-to-grid mode. Finally, the validity of theoretical considerations and control algorithms of some of the existing charging solutions is experimentally verified and experimental performance of all discussed topologies is compared

Additional file 2: of Phylogenetic, syntenic, and tissue expression analysis of slc22 genes in zebrafish (Danio rerio)

Gene structure analysis of zebrafish and human slc22/Slc22. Structure of coding exons, exons and introns together with gene lengths. (XLSX 539 kb

Additional file 3: of Phylogenetic, syntenic, and tissue expression analysis of slc22 genes in zebrafish (Danio rerio)

Transcription factor binding motif analysis of zebrafish and human slc22/Slc22 genes. Identification of potential sequence motifs and regulatory elements of human and zebrafish Slc22/slc22 genes. (XLSX 55 kb

Additional file 1: Figure S1. of Phylogenetic, syntenic, and tissue expression analysis of slc22 genes in zebrafish (Danio rerio)

Extended phylogenetic tree of Slc22/slc22 protein family in vertebrates; Figure S2: Conserved synteny analysis of less characterized human and zebrafish Slc22/slc22 genes; Figure S3: Tissue expression profile of major zebrafish slc22 genes organized by individual genes; Table S1: Protein annotations with accession numbers of all analysed species; Table S2: Heatmap of zebrafish Slc22 gene tissue expression results. (XLSX 1109 kb

Additional file 4: of Phylogenetic, syntenic, and tissue expression analysis of slc22 genes in zebrafish (Danio rerio)

Amino acid sequence motif analysis of zebrafish and human Slc22/SLC22. Identification of characteristic Slc22 amino acid sequences with identified major facilitator superfamily (MFS) motif, amphiphilic solute facilitator (ASF) domain and OAT-specific motif. (XLSX 518 kb

Propofol Induces Apoptosis of Neurons but Not Astrocytes, Oligodendrocytes, or Neural Stem Cells in the Neonatal Mouse Hippocampus

It has been shown that propofol can induce widespread apoptosis in neonatal mouse brains followed by long-term cognitive dysfunction. However, selective brain area and cell vulnerability to propofol remains unknown. This study was aimed to dissect toxic effect of propofol on multiple brain cells, including neurons, astrocytes, oligodendrocytes, and neural stem cells (NSCs). Seven-day-old mice were intraperitoneally administrated propofol or intralipid as a vehicle control for 6 hours. To identify vulnerable cells undergoing apoptosis following propofol exposure, brain sagittal sections were co-stained with antibodies against an apoptosis marker along with neuron, astrocyte, oligodendrocyte, or NSC markers using immunofluorescence staining. The results showed widespread apoptosis in propofol-treated brains (apoptotic cells: 1.55 ± 0.04% and 0.06 ± 0.01% in propofol group and intralipid-treated control group, respectively). Apoptotic cell distribution exhibits region- and cell-specific patterns. Several brain regions (e.g., cerebral cortex and hippocampus) were more vulnerable to propofol than other brain regions. Most apoptotic cells in the hippocampus were located in the cornus ammonis 1 (CA1) subfield. These apoptotic cells were only detected in neurons and not astrocytes, oligodendrocytes, or NSCs. These data demonstrate that different brain regions, subfields, and different types of neuronal cells in mice exhibit various vulnerabilities to propofol. Understanding region- and cell-specific susceptibility to propofol will help to better understand cellular contribution to developmental neurotoxicity and further develop novel therapeutic targets

Identifying Family-Member-Specific Targets of Mono-ARTDs by Using a Chemical Genetics Approach

ADP-ribosyltransferases (ARTD1–16) have emerged as major downstream effectors of NAD+ signaling in the cell. Most ARTDs (ARTD7 and 8, 10–12, and 14–17) catalyze the transfer of a single unit of ADP-ribose from NAD+ to target proteins, a process known as mono-ADP-ribosylation (MARylation). Progress in understanding the cellular functions of MARylation has been limited by the inability to identify the direct targets for individual mono-ARTDs. Here, we engineered mono-ARTDs to use an NAD+ analog that is orthogonal to wild-type ARTDs. We profiled the MARylomes of ARTD10 and ARTD11 in vitro, identifying isoform-specific targets and revealing a potential role for ARTD11 in nuclear pore complex biology. We found that ARTD11 targeting is dependent on both its regulatory and catalytic domains, which has important implications for how ARTDs recognize their targets. We anticipate that our chemical genetic strategy will be generalizable to all mono-ARTD family members based on the similarity of the mono-ARTD catalytic domains