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Changing Viral Tropism Using Immunoliposomes Alters the Stability of Gene Expression: Implications for Viral Vector Design

Changing Viral Tropism Using Immunoliposomes Alters the Stability of Gene Expression: Implications for Viral Vector Design

Since the initial description of lipofec- tion as a mean of DNA transfection (39), cationic liposomes have been widely used for gene delivery both in vitro and in vivo (40,41). We, like others, have used liposomes to package viral particles to increase the infectivity (17,18,42). Using immunoliposomes targeted to endothe- lial receptors, such as CD71 (nonacti- vated ECs) or activated endothelial re- ceptors, such as CD62E/P, we can enhance gene transfer of MMLV. We showed selectivity of viral tropism when viral particles were complexed with im- munoliposomes containing anti-CD71 antibodies. Although expression of CD71 is high on ECs, its ubiquitous expression restricts its use as a specific target for gene delivery. We have, therefore, devel- oped this approach with anti-CD62E/P. This allows a target-specific delivery of viral particles to activated ECs. On the other hand, other adhesion molecules, such as CD54 (data not shown) and CD106 expressed by activated ECs (21), did not show enhancement of transduc- tion efficiency. Our observations suggest that the cellular receptors chosen to be targeted are important in determining the success of the receptor-mediated ap- proach. Whereas some antigens may be attractive targets in terms of patterns of expression, they are poor at delivering the viral particles to the appropriate cel- lular compartment. For example, even though CD106 has been shown to inter- nalize through clathrin-coated vesicles (43), when the molecule is bound by an- tibody, it is shed rather than internalized (44). CD54, on the other hand, has a 10-fold lower endocytosis rate than E-selectin (19,45).
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HIV 1 subtype and viral tropism determination for evaluating antiretroviral therapy options: an analysis of archived Kenyan blood samples

HIV 1 subtype and viral tropism determination for evaluating antiretroviral therapy options: an analysis of archived Kenyan blood samples

Background: Infection with HIV-1 is characterized by genetic diversity such that specific viral subtypes are predominant in specific geographical areas. The genetic variation in HIV-1 pol and env genes is responsible for rapid development of resistance to current drugs. This variation has influenced disease progression among the infected and necessitated the search for alternative drugs with novel targets. Though successfully used in developed countries, these novel drugs are still limited in resource-poor countries. The aim of this study was to determine HIV-1 subtypes, recombination, dual infections and viral tropism of HIV-1 among Kenyan patients prior to widespread use of antiretroviral drugs.
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Generation of recombinant adenovirus vectors with modified fibers for altering viral tropism.

Generation of recombinant adenovirus vectors with modified fibers for altering viral tropism.

To expand the utility of recombinant adenovirus vectors for gene therapy applications, methods to alter native viral tropism to achieve cell-specific transduction would be beneficial. To this end, we are pursuing genetic methods to alter the cell recognition domain of the adenovirus fiber. To incorporate these modified fibers into mature virions, we have developed a method based on homologous DNA recombination between two plasmids. A fiber-deleted, propagation-defective rescue plasmid has been designed for recombination with a shuttle plasmid encoding a variant fiber gene. Recombination between the two plasmids results in the derivation of recombinant viruses containing the variant fiber gene. To establish the utility of this method, we constructed a recombinant adenovirus containing a fiber gene with a silent mutation. In addition, we generated an adenovirus vector containing chimeric fibers composed of the tail and shaft domains of adenovirus serotype 5 and the knob domain of serotype 3. This modification was shown to alter the receptor recognition profile of the virus containing the fiber chimera. Thus, this two-plasmid system allows for the generation of adenovirus vectors containing variant fibers. This method provides a rapid and facile means of generating fiber-modified recombinant adenoviruses. In addition, it should be possible to use this system in the development of adenovirus vectors with modified tropism to allow cell-specific targeting.
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Arterivirus Minor Envelope Proteins Are a Major Determinant of Viral Tropism in Cell Culture

Arterivirus Minor Envelope Proteins Are a Major Determinant of Viral Tropism in Cell Culture

Arteriviruses can cause various clinical symptoms ranging from lethal to asymptomatic persistent infection in susceptible host an- imals (42). Of the four family members, EAV and PRRSV stand out for economic reasons, and the latter virus has rapidly become one of the most economically important swine pathogens world- wide. Furthermore, EAV and PRRSV have attracted attention due to a variety of molecular biological features and have been devel- oped into important models for basic research on arteriviruses and nidoviruses at large. Entry is the first step of the viral replica- tive cycle, and dissection of this process will be of great significance for controlling arterivirus infection. However, the molecular in- teractions involved in arterivirus attachment and entry have re- mained poorly understood and “controversial,” to say the least. It is generally accepted that PRRSV enters host cells via receptor- mediated endocytosis (22, 49). Several cellular molecules have been proposed as potential cellular receptors, including heparan sulfate, pSn, CD151, CD163, and vimentin (1, 13, 21, 40, 51). Of these cellular molecules, pSn has been most extensively studied, but its role was challenged by the discovery of another potential receptor, CD163, a macrophage-specific protein in the scavenger
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A stable CC chemokine receptor (CCR) 5 tropic virus is correlated with the persistence of HIV RNA at less than 2 5 copies in successfully treated naïve subjects

A stable CC chemokine receptor (CCR) 5 tropic virus is correlated with the persistence of HIV RNA at less than 2 5 copies in successfully treated naïve subjects

harbouring an R5- or X4-tropic virus. This result confirms those obtained in previous studies showing no differences in the virological treatment response between patients harbouring an X4- or R5-tropic virus, although a poor recovery of CD4 cells has been shown in patients with an X4-tropic virus at baseline or after an R5 to X4 virus switch [7,11-13,24]. Nevertheless, in our cohort the mean CD4 count increased in the interval T1 to T2, and this increase was significantly higher patients harbouring R5 coreceptor tropism than for X4 tropism (p = 0.0497). This finding suggests a possible difference in the long- term course of this disease. Some biases of the study should be considered. First of all, since viral DNA sequenced from PBMCs may be a reflection of HIV archived at different stages of infection, changes in viral tropism detected in our study may be caused by a sam- pling issue or by technical reasons in consequence of the use of population sequencing rather than viral evolution. To avoid or minimize these biases we repeated twice all sequences with a false positive rate surrounding the cut- off values. Additionally, it was not possible to obtain an analysis of the tropism for all patients at all time points studied. However, this study analysed numerous viro- immunological parameters, and a large number of patients was evaluated at individual time points. In 51 patients, it was possible to evaluate the various parameters through- out the course of treatment.
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umi-unc-2665.pdf

umi-unc-2665.pdf

Mosaic capsid modification can also expand viral tropism because it involves mixing capsid subunits from different serotypes to create a hybrid capsid with receptor binding profiles conferred from each parent serotype (reviewed in (Wu, Asokan et al. 2006). It was shown that mosaic vectors can gain function compared to the parent serotypes (Rabinowitz, Rolling et al. 2002; Rabinowitz, Bowles et al. 2004). An AAV1/AAV2 mosaic vector where the capsid input ratio was 1 to 1 was compared to an AAV2 vector after hippocampal delivery. The AAV2 vector transduced only the hilar interneurons while the mosaic vector transduced the hilar interneurons and some pyramidal and granule cells (Richichi, Lin et al. 2004). Although higher transduction efficiency was achieved by using the mosaic vector, the authors did not include an AAV1 vector in this study. Therefore, it cannot be determined if the mosaic vector would outperform each parent serotype. In addition, this method of viral production actually yields non-uniform virions. For example, even though in this case an equal ratio of type 1 and type 2 helper plasmids were used, it is likely that not all particles will contain equal amounts of type 1 and type 2 subunits. Since standardization of these preps is difficult, this method of capsid engineering will probably not transition to clinical trials.
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TROCAI (Tropism Coreceptor Assay Information): a New Phenotypic Tropism Test and Its Correlation with Trofile Enhanced Sensitivity and Genotypic Approaches

TROCAI (Tropism Coreceptor Assay Information): a New Phenotypic Tropism Test and Its Correlation with Trofile Enhanced Sensitivity and Genotypic Approaches

Different genotypic and phenotypic tools have been used to assay HIV tropism (2). Despite the improvements, genotypic algorithms are not sensitive enough, and their predictive values have limitations (7, 14, 16). Another genotypic approach is ultradeep sequencing; however, this is currently expensive and not applicable to daily clinical practice (1). Two different com- mercially available phenotypic assays allow the determination of viral tropism: the tropism recombinant test (TRT) (VIRalliance, France) (24) and the original phenotypic test Trofile (Monogram Biosciences, South San Francisco, CA) (27). Trofile is the only clinically validated tropism test; how- ever, it has some limitations, such as (i) high cost, (ii) pro- longed time to obtain a confirmed result, (iii) availability (sam- ples need to be sent to South San Francisco, CA), (iv) a considerable proportion of “nonreportable” results, (v) the
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Mode of antiviral action of silver nanoparticles against HIV-1

Mode of antiviral action of silver nanoparticles against HIV-1

Finally, we propose that the antiviral activity of silver nanoparticles results from their inhibition of the interac- tion between gp120 and the target cell membrane recep- tors. According to our results, this mode of antiviral action allows silver nanoparticles to inhibit HIV-1 infec- tion regardless of viral tropism or resistance profile, to bind to gp120 in a manner that prevents CD4-depen- dent virion binding, fusion, and infectivity, and to block HIV-1 cell-free and cell-associated infection, acting as a virucidal agent. In conclusion, silver nanoparticles are effective virucides as they inactivate HIV particles in a short period of time, exerting their activity at an early stage of viral replication (entry or fusion) and at post- entry stages. The data presented here contribute to a new and still largely unexplored area; the use of nano- materials against specific targets of viral particles.
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Baculovirus-Encoded Protein BV/ODV-E26 Determines Tissue Tropism and Virulence in Lepidopteran Insects

Baculovirus-Encoded Protein BV/ODV-E26 Determines Tissue Tropism and Virulence in Lepidopteran Insects

Lepidopteran nucleopolyhedroviruses (NPVs) show distinct tissue tropism in host insect larvae. However, the molecular mecha- nism of this tropism is largely unknown. We quantitatively investigated NPV tissue tropism by measuring mRNA levels of viral genes in 16 tissues from Bombyx mori NPV (BmNPV)-infected B. mori larvae and found clear tissue tropism, i.e., BmNPV repli- cates poorly in the silk glands, midgut, and Malpighian tubule compared with other larval tissues. We next identified the viral genes determining tissue tropism in NPV infection by investigating the phenotypes of larvae infected with 44 BmNPV mutants in which one gene was functionally disrupted by a LacZ cassette insertion. We found that occlusion body (OB) production was markedly enhanced compared with that of the wild type in the middle silk glands (MSGs) of larvae infected with three mutants in which one of three tandemly arrayed genes (Bm7, Bm8, and Bm9) was disrupted. We generated additional mutants in which one or two genes of this gene cluster were partially deleted and showed that Bm8, also known as BV/ODV-E26, was solely re- quired for the suppression of OB production in the MSGs of BmNPV-infected B. mori larvae. Western blotting showed that a LacZ cassette insertion in Bm7 or Bm9 resulted in aberrant expression of Bm8, presumably leading to abnormal OB production in the MSGs. Larval bioassays also revealed that disruption of Bm8 accelerated the death of B. mori larvae. These results suggest that the group I NPV-specific protein BV/ODV-E26 determines tissue tropism and virulence in host lepidopteran insects.
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Evolution of the Equine Infectious Anemia Virus Long Terminal Repeat during the Alteration of Cell Tropism

Evolution of the Equine Infectious Anemia Virus Long Terminal Repeat during the Alteration of Cell Tropism

levels were about five times higher than that reported for HIV in a single-cycle analysis (29, 31). However, it should be noted that genetic variation studies on the equivalent region of the HIV genome have not been extensively explored and levels of variation can be influenced by the region of the genome under study (24). Concomitant with a burst of genetic diversification in macrophages, a C-to-T change upstream from the enhancer region in the LTR U3 was fixed as the predominant sequence within the population. This change had no detectable effect on LTR transcription in our reporter gene studies. Almost no variation was detected in macP10 sequences, but variation slowly increased over the remaining passages in UVECs and ED cells. The slow increase in divergence during these pas- sages suggested random genetic drift of a small population (19). As the viral stock was passaged through UVECs and fibroblasts that established a broader cell tropism, two more nucleotide changes were fixed within the LTR U3 population that created a new transcription factor binding motif that en- hanced LTR activity in fibroblasts but not macrophages. The generation of a new motif within the adapting LTR has been previously demonstrated in HIV (54). A final mutation found in the last passage in fibroblast was a G-to-C change immedi- ately upstream from a PU.1 site that has been reported earlier in fibroblast-tropic strains of virus (43). While the overall con- sensus changes observed in the LTR were modest, some of the genotypic changes that became predominant in the population resulted in phenotypic changes that correlated well with the observed cell tropism alterations.
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HIV 1 macrophage tropism is determined at multiple levels of the viral replication cycle

HIV 1 macrophage tropism is determined at multiple levels of the viral replication cycle

Here, using primary HIV-1 isolates, we demonstrate that differences in macrophage-tropism as observed between absolute macrophage-tropic and non-macrophage-tropic isolates indeed might b[r]

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ppat.0030005.pdf

ppat.0030005.pdf

The mouse-adapted SARS-CoV MA15 virus will be a valuable tool in evaluating SARS-CoV vaccines and antiviral therapy. Quantitative virology was the only outcome measure available in young BALB/c mice challenged with SARS-CoV (Urbani). Because the MA15 virus replicates rapidly to high titer and is lethal for young BALB/c mice, this virus provides a more stringent challenge than the SARS-CoV (Urbani) virus in evaluating the efficacy of therapeutic interventions or prevention strategies. Prophylaxis that is able to rescue MA15-infected mice from a lethal outcome must lower viral burden in lungs very rapidly (e.g., within the first 24 h). Prophylaxis that accomplishes such reduction in the burden of MA15 virus may also reduce the severity of immunopa- thology that follows. A reduction of immunopathology was demonstrated in a SARS-CoV hamster model when a monoclonal antibody specific to SARS-CoV spike protein administered the day after infection was able to arrest a further rise in viral titer in the lungs [29]. If similar protection can be demonstrated against challenge with MA15 virus in BALB/c mice, the efficacy of intervention will be well proven. Infection of young BALB/c mice with the MA15 virus provides a model that is small and accessible and that can be evaluated extensively at an immunological level. Finally, and most importantly, MA15 virus infection of young BALB/c mice provides many elements that replicate observations in acute (and chronic) cases of SARS infection in humans, including sequence changes during adaptation in several genes (nsp 5, nsp 13, S, and M); viral replication and histopathological changes in lungs of infected animals; viremia; detection of vRNA in extrapulmonary sites, includ- ing the intestines; clinical indicators of illness, including mortality; and changes in blood counts, including lympho- penia and neutrophilia. The availability of a molecular clone of the MA15 virus, and the ability to generate additional SARS-CoV recombinant viruses that recapitulate the in vivo phenotype, will allow for detailed probing of the mechanisms mediating SARS-CoV pathogenesis and acute lung damage.
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Coreceptor Tropism in Human Immunodeficiency Virus Type 1 Subtype D: High Prevalence of CXCR4 Tropism and Heterogeneous Composition of Viral Populations

Coreceptor Tropism in Human Immunodeficiency Virus Type 1 Subtype D: High Prevalence of CXCR4 Tropism and Heterogeneous Composition of Viral Populations

Our analysis of HIV env clones revealed significant variabil- ity in the composition of the viral populations from DM-tropic subtype D samples. Samples from individual patients con- tained various mixtures of R5-tropic, X4-tropic, and dual- tropic viruses. Clones that used CXCR4 exclusively were present in most DM samples tested. Furthermore, two differ- ent types of dual-tropic env clones were identified in the virus populations: (i) “dual-X” clones that used CXCR4 efficiently and CCR5 less efficiently (these clones had V3 loop sequences that resembled those from X4-tropic clones from the same sample) and (ii) “dual-R” clones that used CCR5 efficiently and CXCR4 less efficiently (these clones had V3 loop se- quences similar or identical to those from R5-tropic clones from the same sample). Identification of these genetically and phenotypically distinct categories of dual-tropic HIV-1 variants demonstrates that some determinants for CXCR4-usage reside outside of the V3 loop. These determinants may play an im- portant evolutionary role as HIV-1 variants evolve from R5- tropic to X4-tropic. We speculate that the dual-R-tropic vari- ants represent early progenitors of CXCR4-using variants that arise later and use the CXCR4 coreceptor efficiently. We hy- pothesize that mutations within the V3 region that are neces- sary to confer efficient CXCR4 use are highly detrimental to the fitness of CCR5-using strains and are under strong negative selective pressure (Fig. 6). Consequently, in the absence of compensatory changes, the genetic barrier for coreceptor
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Role of Sialic Acid Binding Specificity of the 1918 Influenza Virus Hemagglutinin Protein in Virulence and Pathogenesis for Mice

Role of Sialic Acid Binding Specificity of the 1918 Influenza Virus Hemagglutinin Protein in Virulence and Pathogenesis for Mice

mutation of the HA genes blocked infection or replication. However, we observed dramatic differences in mouse virulence and pathogenesis. Given that the 1918 reassortant viruses were isogenic, apart from the surface glycoproteins, these properties can be attributed to the different HA and NA genes (Table 1). Additionally, the 1918 and avian HA genes were isogenic out- side single nucleotide changes in the codons of the RBD. The parental NY312 virus showed limited replication in the lung at 4 dpi and caused only slight weight loss with minimal patho- logical changes. Replacing the parental NY312 virus genes with wild-type avian HA and NA genes slightly increased weight loss but was still nonfatal. Changing avian HA ␣ 2-3 binding specificity to ␣ 2-6 led to prolonged viral replication of the Av-mut virus in the mouse lung, but with minimal patho- FIG. 4. Pathology and immunohistochemistry of 1918 HA-expressing virus-infected mouse lung tissue. Photomicrographs of hematoxylin-and- eosin-stained tissue sections and immunohistochemically stained sections to detect influenza viral antigen from mice infected with different influenza virus constructs at 6 dpi. Viral antigen is stained red-brown on a hematoxylin-stained background. Arrows show examples of positive cells. (A to C) Sections from an animal infected with the 1918-SC virus. Moderate-to-marked acute alveolitis and bronchiolitis were seen (A, original magnification, ⫻ 20; B, original magnification, ⫻ 40). Viral antigen was observed in alveolar lining cells and in the epithelium of terminal bronchioles (C, original magnification, ⫻ 20). (D to F) Sections from an animal infected with the 1918-NY virus. Moderate-to-marked acute alveolitis and bronchiolitis were seen (D, original magnification, ⫻ 20; E, original magnification, ⫻ 40). Viral antigen was observed in alveolar lining cells (F, original magnification, ⫻ 40). (G to I) Sections from an animal infected with the 1918-NYmut virus. Moderate acute alveolitis and bronchiolitis were seen (G, original magnification, ⫻ 20; H, original magnification, ⫻ 40). Viral antigen was observed in alveolar lining cells (I, original magnification, ⫻ 20).
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Protein pUL128 of Human Cytomegalovirus Is Necessary for Monocyte Infection and Blocking of Migration

Protein pUL128 of Human Cytomegalovirus Is Necessary for Monocyte Infection and Blocking of Migration

HCMV uses monocytes as an important target of infection, subverting their roles in both innate and adaptive immune responses. In fact, monocytes are regarded as a site of viral latency in vivo (26, 30) and as vehicles for viral dissemination (28, 32, 35). At present, little is known about the mechanisms of HCMV entry and establishment of infection in these cells or about the influence of HCMV on their functions. Monocytes originate from CD34 ⫹ progenitors in the bone marrow, circu- late in the bloodstream for 1 to 3 days, and then move into peripheral tissues in order to replenish the resident popula- tions of tissue macrophages and dendritic cells. Monocytes can encounter HCMV during all of these stages: in the bone mar- row, in the bloodstream, and in the peripheral tissues during or after transendothelial migration from the bloodstream. Mono- cyte movements are tightly controlled by members of the su- perfamily of chemoattractant cytokines called chemokines and by their receptors (22). The deregulation of the chemokine- chemokine receptor system can favor HCMV infection, spread, and persistence in the host. Accordingly, more than 30 distinct virally encoded proteins have been identified that are able to corrupt the chemokine system (15); in particular, HCMV encodes two CXC chemokine homologues (18) and four chemokine receptor homologues (36).
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Differential Cell Tropism of Feline Immunodeficiency Virus Molecular Clones In Vivo

Differential Cell Tropism of Feline Immunodeficiency Virus Molecular Clones In Vivo

Previous studies assessing FIV-pF34, FIV-14, and FIV- pPPR have shown these viruses to be unique for both in vitro growth properties and replicative efficiency in vivo (23, 25, 27, 29, 30). However, experimental studies comparing virus repli- cation and host cell tropism of all three cloned isolates in primary cell culture systems, as well as in cats, have not been reported. For in vitro studies, replication of these molecular clones was compared in CrFK cells, a feline adherent cell line, primary feline PBMC, and primary feline MDM. CrFK cells were permissive for both FIV-14 and FIV-pF34 and nonper- missive for FIV-pPPR. Primary feline PBMC were permissive for FIV-pPPR and FIV-14, whereas replication of FIV-pF34 was either severely restricted or absent. These observations are in agreement with previous reports (23, 25, 32, 40) and can be explained by the origin of the viruses and known tropism de- terminants (34, 38, 39).
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Retargeting of Rat Parvovirus H-1PV to Cancer Cells through Genetic Engineering of the Viral Capsid

Retargeting of Rat Parvovirus H-1PV to Cancer Cells through Genetic Engineering of the Viral Capsid

P arvoviridae are small, nonenveloped, single-stranded DNA vi- ruses that infect a wide variety of animal species, from insects to humans (60). Rodent members of the genus Parvovirus (PV), such as minute virus of mice (MVM) and rat H-1PV, attract high levels of interest as novel anticancer agents, because they can rep- licate autonomously in oncogene-transformed cells and exert both oncolytic and oncosuppressive activities in various cell cul- ture and animal models while being nonpathogenic for humans (41, 57). The oncoselectivity of PVs is not due to better virus uptake by transformed cells but to a more efficient viral replica- tion and/or toxicity in these cells. This results in part from the fact that PV DNA replication and gene expression are dependent on cellular factors such as E2F, CREB, ATF, cyclin A (57), and others, all of which are known to be upregulated in cancer cells. More- over, in contrast to normal cells, cancer cells are unable to mount an efficient antiviral defense against PV (22), thus providing more favorable conditions for the viral life cycle. Besides their antineo- plastic activities, another advantage of rodent PVs for cancer ther- apy is the lack of previous exposure of (most) humans to these agents, precluding the rapid elimination of the virus inoculum through preexisting antiviral immunity (11). Taken together, these properties make these viruses very attractive candidates for use as anticancer agents.
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Relationship of Facet Tropism with Degeneration and Stability of Functional Spinal Unit

Relationship of Facet Tropism with Degeneration and Stability of Functional Spinal Unit

facet joint tropism was a predisposing factor for the development of degenerative spondylolisthesis. The present study found no association between facet tropism and translational segmental motion (such as vertebral slippage) within the lumbar spine. Our results indicate that facet tropism has no major association with the deve- lopment of degenerative spondylolisthesis.

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Identification of a Novel Hepacivirus in Domestic Cattle from Germany

Identification of a Novel Hepacivirus in Domestic Cattle from Germany

I t is estimated that over 185 million people are infected with hepatitis C virus (HCV) worldwide (1). HCV represents the type species of the genus Hepacivirus within the family Flaviviridae, which also includes the genera Flavivirus, Pestivirus, and Pegivirus. Although treatment options will be significantly expanded in coming years, most patients living in developing countries will not profit from novel drug therapies, and protective vaccines are still not available to date (2, 3). The development of several rodent models has only partially overcome the challenges to studying HCV in vivo (4). Alternatively, a relative of HCV, the GB virus B (GBV-B), is considered a surrogate HCV infection model (5, 6). In recent years, it became apparent that hepaciviruses are more wide- spread than originally suspected: the identification of HCV-like sequences in wild and domestic animals broadened the spectrum of alternative animal models. In 2011, an HCV-like virus was found in dogs (7). However, serology and PCR-based studies re- vealed that the natural reservoirs are not dogs but horses. Subse- quently, these sequences were designated nonprimate hepacivi- ruses (NPHV) (8, 9). Moreover, a great diversity of hepacivirus sequences was detected in rodents and bats from Europe, Africa, Asia, and Central America (10, 11). In addition to HCV-like se- quences, other viral genomes assigned to the proposed genus Pegi- virus were identified, expanding the family Flaviviridae (11–16).
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Identification of a potential mechanism of acute kidney injury during the COVID-19 outbreak: a study based on single-cell transcriptome analysis

Identification of a potential mechanism of acute kidney injury during the COVID-19 outbreak: a study based on single-cell transcriptome analysis

[9] Meng T, Cao H, Zhang H, Kang Z, Xu D, Gong H, et al. The insert sequence in SARS-CoV-2 enhances spike protein cleavage by TMPRSS. bioRxiv. 2020:doi: https://doi.org/10.1101/2020.02.08.926006. [10] Hoffmann M, Kleine-Weber H, Krüger N, M ü ller M, Drosten C, Pöhlmann S. The novel coronavirus 2019 (2019-nCoV) uses the SARS-coronavirus receptor ACE2 and the cellular protease TMPRSS2 for entry into target cells. bioRxiv. 2020:doi: https://doi-org-tu.vtrus.net/10.1101/2020.01.31.929042. [11] Wan Y, Shang J, Graham R, Baric RS, Li F. Receptor recognition by novel coronavirus from Wuhan: An analysis based on decade-long structural studies of SARS. J Virol. 2020: doi: 10.1128/JVI.00127-20. [12] Gallagher TM, Buchmeier MJ. Coronavirus spike proteins in viral entry and pathogenesis. Virology. 2001;279:371-4.
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