Isolation and characterization of the herpes simplex virus 1 terminase complex

During herpes simplex virus 1 (HSV-1) infection, empty procapsids are assembled and subsequently filled with the viral genome by means of a protein complex called the terminase, which is comprised of the HSV-1 UL15, UL28, and UL33 proteins. Biochemical studies of the terminase proteins have been hampered by the inability to purify the intact terminase complex. In this study, terminase complexes were isolated by tandem-affinity purification (TAP) using recombinant viruses expressing either a full-length NTAP-UL28 fusion protein (vFH476) or a C-terminally truncated NTAP-UL28 fusion protein (vFH499). TAP of the UL28 protein from vFH476-infected cells, followed by silver staining, Western blotting, and mass spectrometry, identified the UL15, UL28, and UL33 subunits, while TAP of vFH499-infected cells confirmed previous findings that the C terminus of UL28 is required for UL28 interaction with UL33 and UL15. Analysis of the oligomeric state of the purified complexes by sucrose density gradient ultracentrifugation revealed that the three proteins formed a complex with a molecular mass that is consistent with the formation of a UL15-UL28-UL33 heterotrimer. In order to assess the importance of conserved regions of the UL15 and UL28 proteins, recombinant NTAP-UL28 viruses with mutations of the putative UL28 metal-binding domain or within the UL15 nuclease domain were generated. TAP of UL28 complexes from cells infected with each domain mutant demonstrated that the conserved cysteine residues of the putative UL28 metal-binding domain and conserved amino acids within the UL15 nuclease domain are required for the cleavage and packaging functions of the viral terminase, but not for terminase complex assembly

Patient focused registries can improve health, care, and science.

Eugene Nelson and colleagues call for registries of care data to be transformed into patient centred interactive learning systemsThis work is supported by funding from the Robert Wood Johnson Foundation (Grants: #71211 and 72313), the Cystic Fibrosis Foundation (Grant #OCONNO04Q10), and the Crohn’s and Colitis Foundation of America Quality of Care Initiative (Grant #3372). TSM was funded by the Multidisciplinary Clinical Research Center for Musculoskeletal Diseases at Dartmouth (P60 AR-062799, A. Tosteson, PI), sponsored by the National Institute for Arthritis and Musculoskeletal and Skin Diseases of the National Institutes of Health. EE was funded by The Swedish Research Council for Health, Work Life and Welfare (#2014-4238). MDW is supported by a Wellcome Trust Senior Investigator award (WT09789).This is the final version of the article. It first appeared from BMJ Group via https://doi.org/10.1136/bmj.i331

Performance of turbulence and cavitation models in prediction of incipient and developed cavitation

The aim of this article is to assess the impact of turbulence and cavitation models on the prediction of diesel injector nozzle flow. Two nozzles are examined, an enlarged one, operating at incipient cavitation, and an industrial injector tip, operating at developed cavitation. The turbulence model employed includes the re-normalization group k–ε, realizable k–ε and k–ω shear stress transport Reynolds-averaged Navier–Stokes models; linear pressure–strain Reynolds stress model and the wall adapting local eddy viscosity large eddy simulation model. The results indicate that all Reynolds-averaged Navier–Stokes and the Reynolds stress turbulence models have failed to predict cavitation inception due to their limitation to resolve adequately the low pressure existing inside vortex cores, which is responsible for cavitation development in this particular flow configuration. Moreover, Reynolds-averaged Navier–Stokes models failed to predict unsteady cavitation phenomena in the industrial injector. However, the wall adapting local eddy viscosity large eddy simulation model was able to predict incipient and developed cavitation, while also capturing the shear layer instability, vortex shedding and cavitating vortex formation. Furthermore, the performance of two cavitation methodologies is discussed within the large eddy simulation framework. In particular, a barotropic model and a mixture model based on the asymptotic Rayleigh–Plesset equation of bubble dynamics have been tested. The results indicate that although the solved equations and phase change formulation are different in these models, the predicted cavitation and flow field were very similar at incipient cavitation conditions. At developed cavitation conditions, standard cavitation models may predict unrealistically high liquid tension, so modifications may be essential. It is also concluded that accurate turbulence representation is crucial for cavitation in nozzle flows

Inner tegument protein pUL37 of herpes simplex virus type 1 is involved in directing capsids to the trans-Golgi network for envelopment

Secondary envelopment of herpes simplex virus type 1 has been demonstrated as taking place at the trans-Golgi network (TGN). The inner tegument proteins pUL36 and pUL37 and the envelope glycoproteins gD and gE are known to be important for secondary envelopment. We compared the cellular localizations of capsids from a virus mutant lacking the UL37 gene with those of a virus mutant lacking the genes encoding gD and gE. Although wild-type capsids accumulated at the TGN, capsids of the pUL37− mutant were distributed throughout the cytoplasm and showed no association with TGN-derived vesicles. This was in contrast to capsids from a gD−gE− mutant, which accumulated in the vicinity of TGN vesicles, but did not colocalize with them, suggesting that they were transported to the TGN but were unable to undergo envelopment. We conclude that the inner tegument protein pUL37 is required for directing capsids to the TGN, where secondary envelopment occurs

Оценка напряженно-деформированного состояния элементов конструкции шасси с учетом междуэлементных связей

Стаття присвячена дослідженню впливу міжелементних зв’язків в конструкції на її напружено–деформований стан. Методом скінченних елементів проведено аналіз напружено–деформованого стану деталі з урахуванням між елементних зв’язків та без врахування їх впливу. Отримані результати дозволяють якісно оцінити вплив змін на конструкцію.This article is devoted to studying the influence of interelement ties in the construction of its stress-strain state. The method of finite element analyzes of stress-strain state element design of the chassis in two settlement cases. The first simulation model does not account for the relationship of elements and perceives stress direction. In the second calculation model accounted for between elemental connections. Comparing results shows that the distribution of stresses in lugs latches in the calculation methodology for the development of very different character from the stress distribution that we obtain using classical methods by emergence date due to not linear apposition effort. Maximum loads are placed in the area of the spout and valves are 370 MPa in both study models, but the distribution and importance of stress significantly are different. The values of stresses in the transition zone of I-structures in lugs the calculation model taking into account of interelement connections reaches 49 MPa, and the calculation without relationship – 88 MPa. It should also be noted that in the first case, the elements in the zone of transition work on stretching and the second compression. Value tensions in the area eyelet lugs the calculation model taking into account of interelement ties to 21.6 MPa, and the calculation without considering the relationship reaches to 33.9 MPa. It should also be noted that the nature of the deformation eyelet significantly different. The results can serve as input data for rapid assessment of the stress-strain state design of the chassis in the design of new products and changes to existing designs.Статья посвящена исследованию влияния межэлементных связей в конструкции на ее напряженно-деформированное состояние. Методом конечных элементов проведен анализ напряженно-деформированного состояния детали с учетом межэлементных связей и без учета их влияния. Полученные результаты позволяют качественно оценить влияние изменений на конструкцию