bluuebunny/crossref_metadata_2025_split huggingface dataset:
Rethinking Remdesivir: Synthesis, Antiviral Activity and Pharmacokinetics of Oral Lipid Prodrugs
Authors: Schooley Robert T. , Carlin Aaron F. , Beadle James R. , Valiaeva Nadejda , Zhang Xing-Quan , Clark Alex E. , McMillan Rachel E. , Leibel Sandra L. , McVicar Rachael N. , Xie Jialei , Garretson Aaron F. , Smith Victoria I. , Murphy Joyce , Hostetler Karl Y. 
Published: August 2020

ABSTRACTRemdesivir (RDV, GS-5734) is currently the only FDA-approved antiviral drug for the treatment of SARS CoV-2 infection. The drug is approved for use in adults or children 12-years or older who are hospitalized for the treatment of COVID-19 on the basis of an acceleration of clinical recovery for inpatients with this disease. Unfortunately, the drug must be administered intravenously, restricting its use to those requiring hospitalization for relatively advanced disease. RDV is also unstable in plasma and has a complex activation pathway which may contribute to its highly variable antiviral efficacy in SARS-CoV-2 infected cells. Potent orally bioavailable antiviral drugs for early treatment of SARS-CoV-2 infection are urgently needed and several including molnupiravir and PF-07321332 are currently in clinical development. We focused on making simple, orally bioavailable lipid analogs of Remdesivir nucleoside (RVn, GS-441524) that are processed to RVn-monophosphate, the precursor of the active RVn-triphosphate, by a single-step intracellular cleavage. In addition to high oral bioavailability, stability in plasma and simpler metabolic activation, new oral lipid prodrugs of RVn had submicromolar anti-SARS-CoV-2 activity in a variety of cell types including Vero E6, Calu-3, Caco-2, human pluripotent stem cell (PSC)-derived lung cells and Huh7.5 cells. In Syrian hamsters oral treatment with ODBG-P-RVn was well tolerated and achieved therapeutic levels in plasma above the EC90for SARS-CoV-2. The results suggest further evaluation as an early oral treatment for SARS-CoV-2 infection to minimize severe disease and reduce hospitalizations.
Innate IFNγ is essential for systemicChlamydiacontrol while CD4 T cell-dependent IFNγ production is highly redundant in the female reproductive tract
Authors: Mercado Miguel A.B. , Du Wuying , Malaviarachchi Priyangi A. , Gann Jessica I. , Li Lin-Xi 
Published: August 2020

AbstractProtective immunity to the obligate intracellular bacteriumChlamydiais thought to rely on CD4 T cell-dependent IFNγ production. Nevertheless, whether IFNγ is produced by other cellular source duringChlamydiainfection and how CD4 T cell-dependent and -independent IFNγ contribute differently to host resistance has not been carefully evaluated. In this study, we dissect the requirements of IFNγ produced by innate immune cells and CD4 T cells for resolution ofChlamydia muridarumfemale reproductive tract (FRT) infection. AfterC. muridarumintravaginal inoculation, IFNγ-deficient and T cell-deficient mice exhibited opposite phenotypes for survival and bacterial shedding at the FRT mucosa, demonstrating the distinct requirements for IFNγ and CD4 T cells in host defense againstChlamydia. InRag-deficient mice, IFNγ produced by innate lymphocytes (ILCs) accounted for early bacterial containment and prolonged survival in the absence of adaptive immunity. Although group I ILCs are potent IFNγ producers, we found that mature NK cells and ILC1 were not the sole source for innate IFNγ in response toChlamydia. T cell adoptive transfer experiments revealed that WT and IFNγ-deficient CD4 T cells were equally capable of mediating effective bacterial killing in the FRT during the early stage ofChlamydiainfection. Together, our results revealed that innate IFNγ is essential for preventing systemicChlamydiadissemination, whereas IFNγ produced by CD4 T cells is largely dispensable at the FRT mucosa.
Susceptibility of rabbits to SARS-CoV-2
Authors: Mykytyn Anna Z. , Lamers Mart M. , Okba Nisreen M.A. , Breugem Tim I. , Schipper Debby , van den Doel Petra B. , van Run Peter , van Amerongen Geert , Waal Leon de , Koopmans Marion P.G. , Stittelaar Koert J. , van den Brand Judith M.A. , Haagmans Bart L. 
Published: August 2020

AbstractTransmission of severe acute respiratory coronavirus-2 (SARS-CoV-2) between livestock and humans is a potential public health concern. We demonstrate the susceptibility of rabbits to SARS-CoV-2, which excrete infectious virus from the nose and throat upon experimental inoculation. Therefore, investigations on the presence of SARS-CoV-2 in farmed rabbits should be considered.
Designing of epitope-based vaccine from the conserved region of spike glycoprotein of SARS-CoV-2
Authors: Agarwal Vidhu , Varadwaj Pritish , Tiwari Akhilesh 
Published: August 2020

AbstractThe emergence of COVID-19 as a pandemic with a high morbidity rate is posing serious global concern. There is an urgent need to design a suitable therapy or vaccine that could fight against SARS-CoV-2 infection. As spike glycoprotein of SARS-CoV-2 plays a crucial role in receptor binding and membrane fusion inside the host, it could be a suitable target for designing of an epitope-based vaccine. SARS-CoV-2 is an RNA virus and thus has a property to mutate. So, a conserved peptide region of spike glycoprotein was used for predicting suitable B cell and T cell epitopes. 4 T cell epitopes were selected based on stability, antigenicity, allergenicity and toxicity. Further, MHC-I were found from the immune database that could best interact with the selected epitopes. Population coverage analysis was also done to check the presence of identified MHC-I, in the human population of the affected countries. The T cell epitope that binds with the respective MHC-I with highest affinity was chosen. Molecular dynamic simulation results show that the epitope is well selected. This is an in-silico based study that predicts a novel T cell epitope from the conserved spike glycoprotein that could act as a target for designing of the epitope-based vaccine. Further, B cell epitopes have also been found but the main work focuses on T cell epitope as the immunity generated by it is long lasting as compared to B cell epitope.
CRISPR/Cas9-mediated gene editing induces neurological recovery in an A53T-SNCA overexpression rat model of Parkinson’s disease
Authors: Yoon Hyung Ho , Ye Sunghyeok , Lim Sunhwa , Lee Seung Eun , Oh Soo-Jin , Jo Ara , Lee Hawon , Kim Na-Rae , Kim Kyoungmi , Kim Bum-Joon , Lee C. Justin , Nam Min-Ho , Hur Junseok W. , Jeon Sang Ryong 
Published: August 2020

AbstractTo date, no publicly available disease-modifying therapy for Parkinson’s disease has been developed. This can be partly attributed to the absence of techniques forin vivodeletion of the SNCA gene (encoding α-synuclein), which is one of the key players in Parkinson’s disease pathology. In particular, A53T-mutated SNCA (A53T-SNCA) is one of the most studied familial pathologic mutations in Parkinson’s disease. Here we utilized a recently discovered genome editing technique, CRISPR/Cas9, to delete A53T-SNCAin vitroandin vivo. Among various CRISPR/Cas9 systems, SaCas9-KKH with a single guide RNA (sgRNA) targeting A53T-SNCA was packaged into adeno-associated virus. Adeno-associated virus carrying SaCas9-KKH significantly reduced A53T-SNCA levels in A53T-SNCA-overexpressed HEK293T cells, without off-target effects on wild-type SNCA. Furthermore, we tested the technique’sin vivotherapeutic potential in a viral A53T-SNCA overexpression rat model of Parkinson’s disease. Gene deletion of A53T-SNCA significantly prevented the overexpression of α-synuclein, dopaminergic neurodegeneration, and parkinsonian motor symptoms, whereas a negative control without sgRNA did not. Our findings propose CRISPR/Cas9 system as a potential therapeutic tool for A53T-SNCA familial Parkinson’s disease.
MicroRNA-224 Promotes Cell Migration and Invasion by Targeting HOXA5 expression in Hepatocellular Carcinoma
Authors: Liu Yuwu , Lin Chen , Wang Dongmei , Wen Sailan , Wang Junpu , Chen Zhihong , Du Jing , Ning Chao , Feng Deyun 
Published: August 2020

AbstractMicroRNAs (miRNAs) are involved in the regulation of multiple cellular pathways and play a key role in the development and progression of tumor. Based on the cellular function of their targets, miRNAs play the role of oncogenes or tumor suppressor genes. Multiple studies have shown that abnormal expression of miRNAs has close relation with the incidence of HCC, but the mechanism of miRNAs in HCC still needs further research. In the present study, we showed that the overexpression of miR-224 can reduce the mRNA and protein expression of ADAM17 and HOXA5, the silencing of miR-224 can increase the protein expression of ADAM17 and HOXA5. Dual luciferase reporter assays showed that miR-224 can directly regulate the expression of ADAM17 and HOXA5. Importantly, we found that miR-224 positively regulates cell migration and invasion in HCC, miR-224 overexpression can promote the migration and invasion of BEL-7402 cell, and miR-224 silencing can suppress the migration and invasion of BEL-7402 cell. miR-224 overexpression can result in the redistribution of cell cycle, the cell percentage of S phase was increased significantly, the cell percentage of G1 phase was decreased significantly, and there is no noticeable change for the cell percentage of G2 phase. These results demonstrated that it may be exert the function of oncogenes in a particular link of cancer cell growth. In conclusion, these results suggest that miR-224 will become a promising biological target in the treatment strategy of hepatocellular carcinoma (HCC).
Feasibility of phosphoproteomics on leftover samples after RNA extraction with guanidinium thiocyanate
Authors: Rolfs Frank , Piersma Sander R. , Dias Mariana Paes , Jonkers Jos , Jimenez Connie R. 
Published: August 2020

AbstractIn daily practice, different types of biomolecules are usually extracted for large-scale ‘omics’ analysis with tailored protocols. However, when sample material is limited, an all-in-one strategy is preferable. While lysis of cells and tissues with urea is the accepted standard for phosphoproteomic applications, DNA, RNA and proteins can be simultaneously extracted from small samples using acid guanidinium thiocyanate-phenol-chloroform (AGPC). Use of AGPC for mass spectrometry (MS)-based phosphoproteomics has been reported, but not benchmarked. Here we compared urea-with AGPC-based protein extraction, profiling phosphorylations in the DNA damage response pathway after ionizing irradiation of U2OS cells as proof of principle. On average we identified circa 9000 phosphosites per sample with both extraction methods. Moreover, we observed high similarity of phosphosite characteristics (e.g. 94% shared class 1 identifications) and deduced kinase activities (e.g. ATM, ATR, CHEK1/2, PRKDC). AGPC-based sample extraction can thus replace standard cell lysates for phosphoproteomic workflows and may thus be an attractive way to obtain input material for multiple omics workflows, yielding several data types from a single sample.
Dynamics of neural microstates in the VTA-striatal-prefrontal loop during novelty exploration in the rat
Authors: Mishra A. , Marzban N. , Cohen M. X , Englitz B. 
Published: August 2020

AbstractEEG microstates refer to quasi-stable spatial patterns of scalp potentials, and their dynamics have been linked to cognitive and behavioral states. Neural activity at single and multiunit levels also exhibit spatiotemporal coordination, but this spatial scale is difficult to relate to EEG. Here, we translated EEG microstate analysis to triple-area local field potential (LFP) recordings from up to 192 electrodes in rats to investigate the mesoscopic dynamics of neural microstates within and across brain regions.We performed simultaneous recordings from the prefrontal cortex (PFC), striatum (STR), and ventral tegmental area (VTA) during awake behavior (object novelty and exploration). We found that the LFP data can be accounted for by multiple, recurring, quasi-stable spatial activity patterns with an average period of stability of ~60-100 ms. The top four maps accounted for 60-80% of the total variance, compared to ~25% for shuffled data. Cross-correlation of the microstate time-series across brain regions revealed rhythmic patterns of microstate activations, which we interpret as a novel indicator of inter-regional, mesoscale synchronization. Furthermore, microstate features, and patterns of temporal correlations across microstates, were modulated by behavioural states such as movement and novel object exploration. These results support the existence of a functional mesoscopic organization across multiple brain areas, and open up the opportunity to investigate their relation to EEG microstates, of particular interest to the human research community.Significance StatementThe coordination of neural activity across the entire brain has remained elusive. Here we combine large-scale neural recordings at fine spatial resolution with the analysis of microstates, i.e. short-lived, recurring spatial patterns of neural activity. We demonstrate that the local activity in different brain areas can be accounted for by only a few microstates per region. These microstates exhibited temporal dynamics that were correlated across regions in rhythmic patterns. We demonstrate that these microstates are linked to behavior and exhibit different properties in the frequency domain during different behavioural states. In summary, LFP microstates provide an insightful approach to studying both mesoscopic and large-scale brain activation within and across regions.
Dual contributions of cerebellar-thalamic networks to learning and offline consolidation of a complex motor task
Authors: Varani Andres P , Sala Romain W , Mailhes-Hamon Caroline , Frontera Jimena L , Léna Clément , Popa Daniela 
Published: August 2020

SUMMARYThe contribution of cerebellum to motor learning is often considered to be limited to adaptation, a short-timescale tuning of reflexes and previous learned skills. Yet, the cerebellum is reciprocally connected to two main players of motor learning, the motor cortex and the basal ganglia, via the ventral and midline thalamus respectively. Here, we evaluated the contribution of cerebellar neurons projecting to these thalamic nuclei in a skilled locomotion task in mice. In the cerebellar nuclei, we found task-specific neuronal activities during the task, and lasting changes after the task suggesting an offline processing of task-related information. Using pathway-specific inhibition, we found that dentate neurons projecting to the midline thalamus contribute to learning and retrieval, while interposed neurons projecting to the ventral thalamus contribute to the offline consolidation of savings. Our results thus show that two parallel cerebello-thalamic pathways perform distinct computations operating on distinct timescales in motor learning.
A dissemination-prone morphotype enhances extrapulmonary organ entry by the fungusCryptococcus neoformans
Authors: Denham Steven T. , Brammer Brianna , Chung Krystal Y. , Wambaugh Morgan A. , Bednarek Joseph M. , Guo Li , Brown Jessica C.S. 
Published: August 2020

SUMMARYEnvironmental pathogens, which move from ecological niches to mammalian hosts, must adapt to dramatically different environments. Microbes that disseminate farther, including the fungal meningitis pathogenCryptococcus neoformans, require additional adaptation to diverse tissues. WhenC. neoformansenters the lungs, infecting cells (<10 μm diameter) enlarge (>30 μm diameter), then form a heterogeneous population. The brain contains uniformly small cells (∼7 μm). We demonstrate that formation of a smallC. neoformansmorphotype – called “seed” cells due to their disseminating ability – is critical for extrapulmonary organ entry. Seed cell formation is triggered by environmental factors, includingC. neoformans’ environmental niche, pigeon guano. The underlying trigger, phosphate, can be released by tissue damage, potentially establishing a feed-forward loop of seed cell formation and dissemination. We demonstrate thatC. neoformans’ size variation is not just a continuum but inducible subpopulations that change host interactions to facilitate microbe survival and spread.
Cryo-EM structure of the MgtE Mg2+channel pore domain in Mg2+-free conditions reveals cytoplasmic pore opening
Authors: Jin Fei , Sun Minxuan , Fujii Takashi , Yamada Yurika , Wang Jin , Maturana Andrés D. , Wada Miki , Su Shichen , Ma Jinbiao , Takeda Hironori , Kusakizako Tsukasa , Tomita Atsuhiro , Nakada-Nakura Yoshiko , Liu Kehong , Uemura Tomoko , Nomura Yayoi , Nomura Norimichi , Ito Koichi , Nureki Osamu , Namba Keiichi , Iwata So , Yu Ye , Hattori Motoyuki 
Published: August 2020

ABSTRACTMgtE is a Mg2+channel conserved in organisms ranging from prokaryotes to eukaryotes, including humans, and plays an important role in Mg2+homeostasis. The previously determined MgtE structures in the Mg2+-bound, closed state and structure-based functional analyses of MgtE revealed that the binding of Mg2+ions to the MgtE cytoplasmic domain induces channel inactivation to maintain Mg2+homeostasis. However, due to the lack of a structure of the MgtE channel, including its transmembrane domain in Mg2+-free conditions, the pore-opening mechanism of MgtE has remained unclear.Here, we determined the cryoelectron microscopy (cryo-EM) structure of the MgtE-Fab complex in the absence of Mg2+ions. The Mg2+-free MgtE transmembrane domain structure and its comparison with the Mg2+-bound, closed-state structure, together with functional analyses, showed the Mg2+-dependent pore opening of MgtE on the cytoplasmic side and revealed the kink motions of the TM2 and TM5 helices at the glycine residues, which are important for channel activity. Overall, our work provides structure-based mechanistic insights into the channel gating of MgtE.
Crowding out agricultural insurance and the subsidy system in Italy: empirical evidence of the charity hazard phenomenon
Authors: Miglietta Pier Paolo , Porrini Donatella , Fusco Giulio , Capitanio Fabian 
Published: August 2020

PurposeThe term “charity hazard” refers to the issue of the crowding out of insurance by co-existing relief programs in the context of different institutional governmental disaster schemes. In this context, the aim of this paper is to verify if the charity hazard phenomenon exists in the Italian agricultural insurance scheme.Design/methodology/approachAnnual data regarding crop insurance, subsidies and farm structure were extracted from ISMEA, ISTAT and FADN databases. A SYS-GMM dynamic panel model was estimated, considering the 2010–2017 time period and the Italian Regions as units of the analysis.FindingsThe empirical results highlight a negative relation between crop subsidies and the farmers' policies and total premium paid. The disincentive and crowd-out effects of public aid and subsidies on the choice of whether or not to take out an agricultural insurance policy ends up being one of the key factors for the low level of penetration of the agricultural insurance in Italy.Practical implicationsSince the diffusion of agricultural insurance can contribute to the general objective of sustainability and resilience, the implementation of alternative solutions to subsidies could be needed (e.g. the introduction of mandatory insurance against adversities or financial support for a geographically specific insurance tool).Originality/valueInvestigating empirically the determinants of the agricultural insurance policy diffusion among the Italian Regions, this study ensures an original contribution to the scientific progress in the field, demonstrating the existence of charity hazard caused by the public subsidies provision.
Optimizing the scheduling of crew deployments in repetitive construction projects under uncertainty
Authors: Hassan Abbas , El-Rayes Khaled , Attalla Mohamed 
Published: August 2020

PurposeThis paper presents the development of a novel model for optimizing the scheduling of crew deployments in repetitive construction projects while considering uncertainty in crew production rates.Design/methodology/approachThe model computations are performed in two modules: (1) simulation module that integrates Monte Carlo simulation and a resource-driven scheduling technique to calculate the earliest crew deployment dates for all activities that fully comply with crew work continuity while considering uncertainty; and (2) optimization module that utilizes genetic algorithms to search for and identify optimal crew deployment plans that provide optimal trade-offs between project duration and crew deployment plan cost.FindingsA real-life example of street renovation is analyzed to illustrate the use of the model and demonstrate its capabilities in optimizing the stochastic scheduling of crew deployments in repetitive construction projects.Originality/valueThe original contribution of this research is creating a novel multiobjective stochastic scheduling optimization model for both serial and nonserial repetitive construction projects that is capable of identifying an optimal crew deployment plan that simultaneously minimizes project duration and crew deployment cost.
Linking improvisational behavior, adaptive selling behavior and sales performance
Authors: Charoensukmongkol Peerayuth , Suthatorn Pornprom 
Published: August 2020

PurposeThis research examined the relationships between improvisational behavior, adaptive selling behavior and sales performance of direct sellers in Thailand. This research also investigated whether these relationships are moderated by the degree of challenge orientation and sellers' knowledge about the products.Design/methodology/approachThe data were collected through a survey with sellers from a subsidiary of a multinational corporation located in Bangkok, Thailand (n = 172). Partial least squares–structural equation modeling was used to analyze the data.FindingsResults from data analysis revealed positive relationships between improvisational behavior, adaptive selling behavior and sales performance. The relationship between improvisational behavior and adaptive selling behavior, as well as the relationship between adaptive selling behavior and sales performance, significantly depended on the degree of challenge orientation and the sellers' knowledge about the products.Research limitations/implicationsThe data were collected using self-report measures; the sample was sellers from a single sales organization, and cross-sectional data were used for the analysis. Overall, this study is the exploratory research that does not intend to prove the causal effect of improvisational behavior, but rather to provide new insight on some conditional factors that influence its effectiveness.Practical implicationsIt is essential for sales organizations to ensure that their sales force has adequate improvisational skills to handle sales adaptations effectively during unexpected sales situations. Some training may be offered to the sales force to develop these imperative improvisational skills.Originality/valueThe results regarding the moderating effect of challenge orientation and product knowledge provided additional insight to prior research about the potential conditions that influence the effectiveness of improvisational behavior and adaptive selling behaviors.