EWI-2 Inhibits Cell-Cell Fusion at the HIV-1 Virological Presynapse.

Cell-to-cell transfer of virus particles at the Env-dependent virological synapse (VS) is a highly efficient mode of HIV-1 transmission. While cell-cell fusion could be triggered at the VS, leading to the formation of syncytia and preventing exponential growth of the infected cell population, this is strongly inhibited by both viral (Gag) and host (ezrin and tetraspanins) proteins. Here, we identify EWI-2, a protein that was previously shown to associate with ezrin and tetraspanins, as a host factor that contributes to the inhibition of Env-mediated cell-cell fusion. Using quantitative fluorescence microscopy, shRNA knockdowns, and cell-cell fusion assays, we show that EWI-2 accumulates at the presynaptic terminal (i.e., the producer cell side of the VS), where it contributes to the fusion-preventing activities of the other viral and cellular components. We also find that EWI-2, like tetraspanins, is downregulated upon HIV-1 infection, most likely by Vpu. Despite the strong inhibition of fusion at the VS, T cell-based syncytia do form in vivo and in physiologically relevant culture systems, but they remain small. In regard to that, we demonstrate that EWI-2 and CD81 levels are restored on the surface of syncytia, where they (presumably) continue to act as fusion inhibitors. This study documents a new role for EWI-2 as an inhibitor of HIV-1-induced cell-cell fusion and provides novel insight into how syncytia are prevented from fusing indefinitely

Distinct Regions of the Large Extracellular Domain of Tetraspanin CD9 Are Involved in the Control of Human Multinucleated Giant Cell Formation

Multinucleated giant cells, formed by the fusion of monocytes/macrophages, are features of chronic granulomatous inflammation associated with infections or the persistent presence of foreign material. The tetraspanins CD9 and CD81 regulate multinucleated giant cell formation: soluble recombinant proteins corresponding to the large extracellular domain (EC2) of human but not mouse CD9 can inhibit multinucleated giant cell formation, whereas human CD81 EC2 can antagonise this effect. Tetraspanin EC2 are all likely to have a conserved three helix sub-domain and a much less well-conserved or hypervariable sub-domain formed by short helices and interconnecting loops stabilised by two or more disulfide bridges. Using CD9/CD81 EC2 chimeras and point mutants we have mapped the specific regions of the CD9 EC2 involved in multinucleated giant cell formation. These were primarily located in two helices, one in each sub-domain. The cysteine residues involved in the formation of the disulfide bridges in CD9 EC2 were all essential for inhibitory activity but a conserved glycine residue in the tetraspanin-defining ‘CCG’ motif was not. A tyrosine residue in one of the active regions that is not conserved between human and mouse CD9 EC2, predicted to be solvent-exposed, was found to be only peripherally involved in this activity. We have defined two spatially-distinct sites on the CD9 EC2 that are required for inhibitory activity. Agents that target these sites could have therapeutic applications in diseases in which multinucleated giant cells play a pathogenic role

Loss of PTB or Negative Regulation of Notch mRNA Reveals Distinct Zones of Notch and Actin Protein Accumulation in Drosophila Embryo

Polypyrimidine Tract Binding (PTB) protein is a regulator of mRNA processing and translation. Genetic screens and studies of wing and bristle development during the post-embryonic stages of Drosophila suggest that it is a negative regulator of the Notch pathway. How PTB regulates the Notch pathway is unknown. Our studies of Drosophila embryogenesis indicate that (1) the Notch mRNA is a potential target of PTB, (2) PTB and Notch functions in the dorso-lateral regions of the Drosophila embryo are linked to actin regulation but not their functions in the ventral region, and (3) the actin-related Notch activity in the dorso-lateral regions might require a Notch activity at or near the cell surface that is different from the nuclear Notch activity involved in cell fate specification in the ventral region. These data raise the possibility that the Drosophila embryo is divided into zones of different PTB and Notch activities based on whether or not they are linked to actin regulation. They also provide clues to the almost forgotten role of Notch in cell adhesion and reveal a role for the Notch pathway in cell fusions

Wundballistik in der Rechtsmedizin

Die Beurteilung von Verletzungen durch Schusswaffen gehört zu den typischen Aufgaben der Rechtsmedizin. In den vorangegangenen Kapiteln wurde deutlich dargelegt, dass auf Grund einer Vielzahl zu berücksichtigender physikalischer Parameter und Effekte die rechtsmedizinische Beurteilung derartiger Verletzungen recht komplex ist. Insbesondere spielt die Kurzzeitigkeit der Vorgänge (eine Schussverletzung findet innerhalb einiger weniger Millisekunden statt) eine ganz entscheidende Rolle

Pandora’s box

Suitcases or other containers are occasionally involved in forensic investigations. If there is a suspicion that human remains are hidden inside such a container, medico-legal examinations are required. However, these containers are occasionally locked. Forced opening of a locked suitcase or container may cause damage to its contents. Additionally, the safety of the investigator has to be considered as such containers may be booby-trapped or contain other hazardous material. An overview of the contents before opening is desirable in order to avoid the possibility of encountering a Pandora's box. In forensic medicine, an established approach to examine the inside of a body before opening at autopsy is postmortem computed tomography (CT). However, there may be a reluctance to use this approach for suitcases or containers with metallic components because of the assumption that severe metal artifacts will result in inadequate images. In this article, we present a forensic case in which a CT scan of a metallic suitcase was performed in order to examine its contents. Additionally, we performed an experimental scan of a conventional safe in order to determine if CT is able to reveal the contents of such a highly radiopaque storage box

Fairly direct hit! Advances in imaging of shotgun projectiles in MRI

OBJECTIVES: To investigate the magnetic properties of different types of projectiles and qualify the metal artefact reduction technique for diagnostic and/or forensic MRI. MATERIALS AND METHODS: Ten different projectiles embedded in ordnance gelatine blocks underwent an in vitro 1.5-T MR study with seven sequences including a recently developed metal artefact reduction sequence (Advanced WARP) combining VAT (view-angle-tilting) and SEMAC (slice-encoding metal-artefact-correction). Resulting image quality (five-point scale: 1=best; 5=worst) was scored. Quantifiable magnetic characteristics were correlated with qualitative rating of the MR sequences and torque dislodgment. RESULTS: Metal artefact reduction sequence (median: 2.5) significantly (p < 0.001) improves depiction of projectiles in comparison to all other MR pulse sequences (median: 4.75). Images from diamagnetic composed bullets (median: 2) are much less disturbed compared to magnetic attracted ones (median: 5). Correlation (0.623) between deflection angle measurement (ferromagnetic mean 84.2°; paramagnetic 62°; diamagnetic mean 0°) and median qualitative image quality was highly significant (p = 0.027). Torque dislodgement was distinct for elongated magnetic attracted projectiles. CONCLUSIONS: Significant improvement of MR imaging of projectiles using metal artefact reduction techniques has important implications for diagnostic/forensic work-up. The correlations between magnetic attraction force, deflection-angle results and image properties demonstrate that the MR safety of projectiles can be estimated with one of these methods. KEY POINTS: • Metal artefact reduction sequence improves overall image quality of bullets (p < 0.001). • Deflection angle assessment significantly predicts image quality of bullets (p = 0.027). • Classification of projectiles’ magnetic properties based on artefacts’ characteristics is possible. • Classifying of bullets has important implications in diagnostic and forensic imaging. • Identification of projectiles’ magnetic attributes improves estimation of patients’ injury risk

EWI-2 Inhibits Cell–Cell Fusion at the HIV-1 Virological Presynapse

Cell-to-cell transfer of virus particles at the Env-dependent virological synapse (VS) is a highly efficient mode of HIV-1 transmission. While cell&ndash;cell fusion could be triggered at the VS, leading to the formation of syncytia and preventing exponential growth of the infected cell population, this is strongly inhibited by both viral (Gag) and host (ezrin and tetraspanins) proteins. Here, we identify EWI-2, a protein that was previously shown to associate with ezrin and tetraspanins, as a host factor that contributes to the inhibition of Env-mediated cell&ndash;cell fusion. Using quantitative fluorescence microscopy, shRNA knockdowns, and cell&ndash;cell fusion assays, we show that EWI-2 accumulates at the presynaptic terminal (i.e., the producer cell side of the VS), where it contributes to the fusion-preventing activities of the other viral and cellular components. We also find that EWI-2, like tetraspanins, is downregulated upon HIV-1 infection, most likely by Vpu. Despite the strong inhibition of fusion at the VS, T cell-based syncytia do form in vivo and in physiologically relevant culture systems, but they remain small. In regard to that, we demonstrate that EWI-2 and CD81 levels are restored on the surface of syncytia, where they (presumably) continue to act as fusion inhibitors. This study documents a new role for EWI-2 as an inhibitor of HIV-1-induced cell&ndash;cell fusion and provides novel insight into how syncytia are prevented from fusing indefinitely

Injury potential of thrown sharp kitchen and household utensils

We examined the possibility of inflicting serious injuries with sharp objects in an experimental setting by throwing four sharp objects from different distances and with different throwing techniques. Using an overarm-handle (OA/H), overarm-blade (OA/B), underarm-handle (UA/H), underarm-blade (UA/B) and thrust (T/H) throwing technique, 10 adults (sex ratio 1:1) threw a chef's knife, a skinning knife, a paring knife and office scissors from 4 m and 2 m distance at synthetic abdomen models made of 10% gelatin covered with synthetic skin. The amount of hits and penetrations of the target and their penetration depth were noted, as was the rotation of the blade tip towards the target along its flight trajectory. The chef's knife injury/hit ratio was 0.167/4 m and 0.160/2 m; the skinning knife recorded an injury/hit ratio of 0.087/4 m and 0.153/2 m; the paring knife of 0.087/4 m and 0.113/2 m; and the scissors 0.087/4 m and 0.067/2 m. Mean penetration depths were as follows: the chef's knife: 4 m, 4.42 cm, 2 m, 3.41 cm; the skinning knife: 4 m, 4.19 cm, 2 m, 4.11 cm; the paring knife: 4 m, 1.62 cm, 2 m, 1.55 cm; and the scissors: 4 m, 2.08 cm, 2 m, 0.95 cm. Handle-throw penetration-depths were: 4 m: 3.77 cm and 2 m: 2.86 cm; blade-throw depths were: 4 m: 3.14 cm and 2 m: 2.69 cm. Overarm-throw penetration-depths were: 4 m: 3.62 cm and 2 m: 3.25 cm; and underarm-throw penetration-depths were 4 m: 3.30 cm and 2 m: 2.30 cm. No thrust-throws with the paring knife and scissors could pierce the target. The tips pointed toward the target at angles of 60°-120°, earlier in handle-throws than blade-throws, especially with the paring knife and the scissors. When thrown, especially with a handle-held technique, heavier objects pierced more often and more deeply. Thrust-throws at short distances are unlikely to pierce a human