bluuebunny/crossref_metadata_2025_split huggingface dataset:
Glucose regulated protein 78 (GRP78) interacts with Zika virus envelope and is required for a productive infection
Authors: Royle Jamie , Ramirez-Santana Carolina , Donald Claire , Akpunarlieva Snezhana , Gestuveo Rommel , Anaya Juan-Manuel , Burchmore Richard , Kohl Alain , Varjak Margus 
Published: December 2019

Zika virus (ZIKV) is a member of the Flaviviridae family and was until recently a relatively obscure tropical disease. Subsequently, ZIKV has been shown to be the causative agent of fetal abnormalities and Guillain-Barré syndrome in outbreaks across the Americas and so efforts towards delineating important factors in the viral lifecycle have increased. Combining protein pull-down with mass spectrometry, it was found that ZIKV envelope (Env) interacts with the endoplasmic reticulum (ER) resident chaperone, glucose regulated protein 78 (GRP78) in A549 cells. Flaviviruses such as Japanese encephalitis virus and dengue virus are known to co-opt ER resident proteins and members of the unfolded protein response, including GRP78, to enhance viral infectivity and propagation. The role these proteins play during the ZIKV lifecycle has yet to be elucidated.
To determine the importance of this interaction during ZIKV infection, A549 cells were treated with GRP78-specific siRNAs prior to infection with a NanoLuc expressing reporter virus or a wild-type virus. Depletion of GRP78 significantly reduced both virus luciferase readings and viral titres, indicating that GRP78 is necessary for efficient infection of mammalian cell culture. In contrast, inhibition of GRP78 with small molecule inhibitors did not reduce ZIKV infection. Interestingly, immunofluorescence of ZIKV infected cells reveal that GRP78 re-localises following infection and co-localises with Env. Depletion of GRP78 abrogated localisation of viral replication factories. Further experiments have shown that GRP78 is important for infection post entry and replication, and that putative GRP78 interactions partners are also required during infection.
Identification of candidate molecular determinants of the vector competence of Ixodes ricinus for members of the tick-borne encephalitis complex
Authors: Lemasson Manon , Caignard Grégory , Unterfinger Yves , Attoui Houssam , Bell-Sakyi Lesley , Moutailler Sara , Bonnet Sarah , Vitour Damien , Richardson Jennifer , Lacour Sandrine A. 
Published: December 2019

In Europe, tick-borne encephalitis virus (TBEV) and louping ill virus (LIV) are two flaviviruses both transmitted by the tick Ixodes ricinus. While the mechanisms of viral replication and transmission in this vector are incompletely understood, they are presumed to be largely governed by protein-protein interactions established between viruses and cells.
To elucidate the molecular determinants involved in vector competence, we have mapped the network of protein-protein interactions established between viral proteins of both TBEV and LIV and tick proteins encoded by a cDNA library of I. ricinus, by using yeast two-hybrid methodology. Twenty-two cellular partners from I. ricinus have been identified and all evidenced to interact with both viruses. Upon functional annotation, some of these tick proteins seem to be involved in such biological processes as the immune response or ribosomal maturation. To gain insight into the role of each tick protein in viral replication, the impact of gene silencing will be assessed by dsRNA knockdown. In parallel, I. ricinus cell lines have been persistently infected with TBEV or LIV and the level of expression of selected antiviral effectors monitored over time, in an effort to characterize the antiviral response of this arthropod.
This work represents the first description of the protein-protein interaction network for TBEV, LIV and I. ricinus. Certain tick partners may well represent molecular determinants of vector competence of I. ricinus for TBEV and LIV and potentially other flaviviruses, which we will ascertain by in vivo silencing of selected tick partners.
The Cryopreservation of Cos Lettuce Callus by Using Liquid Nitrogen
Authors: Tangkananond Wipa 
Published: December 2019

The cryopreservation of cos lettuce (CL) callus by using liquid nitrogen (LN) was further study from the previous research on callus induction and micropropagation of CL. That was studied on seed culture and micropropagation of CL and its embryogenic growth. The result was shown that the most suitable Murashing and Skoog (MS) medium concentration was in 1/2 MS. The medium supplemented with 0.1 mg/l of 6-benzylaminopurine (BAP) and 0.5 mg/l of 2,4-dichlorophenoxyacetic acid (2,4-D) gave the best for leaf inducing callus. Then it was brought to study the effects of cryoprotectants with the different percentages of dimethyl sulfoxide (DMSO). The cryopreserved callus by using LN was brought to subculture for the best inducing shoot in MS medium supplemented with 4.0 mg/l of BAP. The results that the best cryoprotectant at the percentage of DMSO at 7.5 gave the highest average number of shoots at 2.1 shoots/culture and the average shoot length was 1.56 cm. The MS medium supplemented with 0.5 mg/l of 1-Naphthaleneacetic acid was the best inducing root. The results that the best cryoprotectant at the percentage of DMSO at 7.5 gave the highest average number of roots at 9.7 roots/culture and the average root length was 1.59 cm. Plant differentiation from callus of CL leaf was transferred to grow in soil and had the same morphology as normal CL that was grown in soil by seed germination.
CL will be the host of Japanese encephalitis virus expression for the developmental swine vaccine.
The effect of co-infecting Anaplasma phagocytophilum on replication of Langat Virus, a model for Louping ill virus, in Ixodes spp. tick cells
Authors: Luu Lisa , Bradley George , Al-Khafaji Alaa , Bell-Sakyi Lesley 
Published: December 2019

Tick borne fever (TBF) caused by Anaplasma phagocytophilum (Ap), and louping ill caused by the flavivirus louping ill virus (LIV) are the two most economically important vector-borne diseases in UK sheep populations. Both pathogens are transmitted by the tick Ixodes ricinus, which also harbours protozoan parasites and Borrelia spp. spirochetes. I. ricinus ticks may be co-infected with multiple microorganisms and potentially transmit more than one pathogen to hosts during blood feeding.
Ap infection is not limited to sheep, causing pasture fever in cattle and granulocytic anaplasmosis in horses, dogs and humans. There is no vaccine available for TBF, and disease control relies on tick control and antibiotic treatment. LIV mainly infects ruminants, but can also infect other livestock including horses, pigs, alpacas and llamas. Since 2016, the LIV vaccine has been unavailable and there is no alternative prophylactic treatment for livestock. Both Ap and LIV are zoonotic diseases with occasional human cases reported in the UK.
Importantly, Ap infection leads to host immunosuppression and consequently increases vulnerability to secondary infections. Both Ap and LIV have been studied as single infections in tick and mammalian cells. However the dynamics and implications of co-infections within the arthropod vector or mammalian host have, to date, not been fully explored.
Using embryo-derived Ixodes spp. cell lines infected with Langat virus (a BSL-2 model for LIV) and Ap, we examined the effect of co-infection on viral RNA replication by qRT-PCR and bacterial growth by qPCR. The results of this study will be presented and discussed.