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    Assessment of replication of bovine herpesvirus type 4 in human glioblastoma and breast cancer cells as a potential oncolytic virus
    (Springer, 2021) Aligholipour Farzani, Touraj; Bilge Dagalp, Seval; Ozkul, Aykut; Gurdal, Hakan; Dogan, Firat; Alkan, Feray
    Oncolytic viruses have been extensively used in cancer treatment due to their tropism, selective replication only in tumor cells, and possible synergic interaction with other therapeutics. Different researchers have demonstrated that bovine herpesvirus 4 (BoHV-4), a member of the gammaherpesviridae family, has oncolytic potential in some human-origin cancer cell lines like glioma through the selective replication strategy. Using four apoptosis detection methods, namely MTT, LDH, TUNEL, and Annexin V assays, we evaluated the apoptotic effect of BoHV-4 Movar33/63 reference strain along with a recombinant BoHV-4 expressing EGFP in U87 MG cells (human glioblastoma cell line), MDA MB-231 (human breast cancer cell line), and MCF10a (non-tumorigenic human mammary epithelial cell line). Our findings indicate that this virus can replicate and induce apoptosis in these cell lines and hinder in vitro proliferation in a dose-dependent manner. In conclusion, BoHV-4 has in vitro potential as a novel oncolytic virus in human cancer therapy. However, its replication potential in the MCF10a cells as a non-tumorigenic human mammary epithelial cell line is a concern in using this virus in cancer therapy, at least against human mammary tumors. Further studies must therefore be conducted to examine the specific apoptotic pathways induced by this virus to move on to further experiments.
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    The genetic diversity of bovine papillomaviruses (BPV) from different papillomatosis cases in dairy cows in Turkey
    (Springer Wien, 2017) Bilge Dagalp, Seval; Dogan, Firat; Farzani, Touraj Alighopour; Salar, Seckin; Bastan, Ayhan
    Papillomaviruses (PVs) are epitheliotropic viruses that cause benign proliferative lesions in the skin (warts or papillomas) and mucous membranes of their natural hosts. In bovines specifically, 13 types of Bovine papillomaviruses (BPVs) are currently described in the literature, although the actual number may be greater than 20. BPV types are classified into four genera based on homology within the genomic regions of the L1 ORF, the most conserved sequence. This study conducted molecular typing of BPV in dairy cows with different papillomatosis cases and investigated the presence of co-infections across distinct BPV types in the same sample. After carrying out PCR using degenerate primers and type specific primers, 35 BPV suspected samples were detected as positive for BPV and these samples were used for typing using sequence analysis/PCR with type-specific primers. This analysis identified BPV-1, -2, -3, -4, -6, -7, -9 and -10, new putative types (BPV/BR/UEL6-like viruses) and the previously described putative type viruses (BAPV-6) in the 35 BPV-positive samples. In addition, co-infections across different BPV types were widely detected in the BPV-positive samples. This study shows that PCR assays using degenerate primers to amplify partial fragments of the L1 gene followed by sequencing is useful for genotyping BPV. However, results need confirmation using type-specific primers in order to consider co-infections. In addition, this study identified a new putative type (in the same cluster as BPV/BR/UEL6-like viruses) and the previously described putative type viruses (BAPV-6) in teat papillomatosis of Turkish dairy cows. The study shows that it is essential to identify BPV types and their prevalence/distribution, and also to determine the clinical consequences of infection for the development of prophylactic and/or therapeutic procedures.
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    Genomic and phylogenetic characterization of ChPV2, a novel goat PV closely related to the Xi-PV1 species infecting bovines
    (Bmc, 2020) Willemsen, Anouk; van den Boom, Alexander; Dietz, Julienne; Bilge Dagalp, Seval; Dogan, Firat; Bravo, Ignacio G.; Ehrhardt, Anja
    Background Papillomaviruses (PVs) infecting artiodactyls are very diverse, and only second in number to PVs infecting primates. PVs associated to lesions in economically important ruminant species have been isolated from cattle and sheep. Methods Potential PV DNA from teat lesions of a Damascus goat was isolated, cloned and sequenced. The PV genome was analyzed using bioinformatics approaches to detect open reading frames and to predict potential features of encoded proteins as well as putative regulatory elements. Sequence comparison and phylogenetic analyses using the concatenated E1E2L2L1 nucleotide and amino acid alignments was used to reveal the relationship of the new PV to the known PV diversity and its closest relevants. Results We isolated and characterized the full-genome of novel Capra hircus papillomavirus. We identified the E6, E7, E1, E2, L2, L1 open reading frames with protein coding potential and putative active elements in the ChPV2 proteins and putative regulatory genome elements. Sequence similarities of L1 and phylogenetic analyses using concatenated E1E2L2L1 nucleotide and amino acid alignments suggest the classification as a new PV type designated ChPV2 with a phylogenetic position within the XiPV genus, basal to the XiPV1 species. ChPV2 is not closely related to ChPV1, the other known goat PV isolated from healthy skin, although both of them belong confidently into a clade composed of PVs infecting cervids and bovids. Interestingly, ChPV2 contains an E6 open reading frame whereas all closely related PVs do not Conclusion ChPV2 is a novel goat PV closely related to the Xi-PV1 species infecting bovines. Phylogenetic relationships and genome architecture of ChPV2 and closely related PV types suggest at least two independent E6 losses within the XiPV clade.
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    Partial sequence of the gB gene of equid herpesvirus type 1 isolates associated with abortion in Turkey
    (Ankara Univ Press, 2016) Ataseven, Veysel Soydal; Oguzoglu, Tuba Cigdem; Dincer, Ender; Bilge Dagalp, Seval
    Equid herpesvirus 1 (EHV-1) is a major agent of large-scale outbreaks of abortion, and these abortions have been described as sporadic or epidemic cases in mares, generally during the last trimester of pregnancy. In this study, the partial characterization based on the glycoprotein B (gB) gene of Turkish (TR) EHV-1 field strains isolated from an abortion outbreak during the 2011 foaling season in Turkey was investigated using a novel designed primer set for EHV-1. The molecular analysis of TR EHV-1 strains showed that genetically identical TR EHV-1 strains were still circulating in Turkey during different years and, these strains were closely related to the European EHV-1 strains. Furthermore, EHV-1 isolated in the present study and to EHV-1 strains published previously in the GenBank database were shown some differences for the aa sequences. This molecular report would be valuable for monitoring of EHV-1 infection in Turkey and determining the gB gene sequence of newly identified EHV-1 field strains from future outbreaks on the pathogenesis and severity of disease.
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    Prevalence of equine herpesvirus-1 and equine herpesvirus-4 infections in equidae species in Turkey as determined by ELISA and multiplex nested PCR
    (Elsevier Sci Ltd, 2009) Ataseven, Veysel S.; Bilge Dagalp, Seval; Guzel, Murat; Basaran, Zeynep; Tan, Mehmet T.; Geraghty, Bob
    In this report we examined the presence of specific antibodies against equine herpesvirus type 1 (EHV-1 and equine herpesvirus type 4 (EHV-4) in several equidae, including mules, donkeys, horses. The presence of EHV-1 and EHV-4 in respiratory diseases of equids, and ability of multiplex nested polymerase chain reaction (PCR) screening in simultaneous diagnosis of horses acutely infected by EHV-1 and EHV-4 were also investigated. Sera from 504 horses, mules and donkeys sampled were tested for the presence of EHV-1 and EHV-4 specific antibodies. Blood samples taken from 21 symptomatic horses and nasal swabs taken from 40 symptomatic horses were tested for the presence of EHV-1 and EHV-4 by a multiplex nested PCR. A total of 14.3% (3/21) of buffy coat samples and 32.5% (13/40) nasal swab samples were found to contain EHV-1 DNA, while 19% (4/21) buffy coat samples and 22.5% (9/40) nasal swab samples were found to be positive for EHV-4 DNA. By species, 14.5% of horses, 37.2% of mules and 24.2% of donkeys tested were EHV-1 seropositive. EHV-4 specific antibodies were detected in 237 (81.7%) of 290 horse sera tested. Results from this investigation demonstrate that EHV-1 and EHV-4 are prevalent throughout the equid population, and that donkeys and mules might also represent an important source of infection for other equids. We also showed that the multiplex nested PCR assay might be useful for diagnosis of mixed respiratory infections in horses due to EHV-1 and EHV-4. (C) 2008 Elsevier Ltd. All rights reserved.
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    A teat papillomatosis case in a Damascus goat (Shami goat) in Hatay province, Turkey: a new putative papillomavirus?
    (Springer Wien, 2018) Dogan, Firat; Dorttas, Selvi Deniz; Bilge Dagalp, Seval; Ataseven, Veysel Soydal; Alkan, Feray
    Papillomaviruses (PVs) are epitheliotropic viruses that cause benign proliferative lesions in the skin (warts or papillomas) and mucous membranes of their natural hosts. Recently, new PVs have been found in many animal species. The most common current approach for identifying novel PV types is based on PCR, using various consensus or degenerated primer (broad-range primers), designed on the basis of the multiple alignment of nucleotide or amino acid sequences of a large number of different human papillomaviruses (HPV). PVs have been classified according to the sequence similarity of one of their capsid proteins, L1, without taking into account other regions of the genome and without considering the phenotypic characteristics of the viral infection. In this study, we performed molecular detection and typing of a PV in a goat with teat papillomatosis. Firstly, PCR was performed using the FAP59/FAP64 and MY09/MY11 primer pairs for the L1 gene region. The PV DNA was found to be positive only with the FAP59/FAP64 primer pair. PV DNA was then tested with three primer sets in four different combinations (L2Bf/FAP64, L2Bf/L1Br, FAP59/FAP64, L1Bf/LCRBr) for the gene region encoding the L1, L2 and LCR proteins. The goat teat papilloma sample was amplified using FAP59/FAP64 primers and two primer pairs (L2Bf/FAP64 and L2Bf/L1Br). We obtained products matching approximately 604 bp of the L1 region of the virus. PV DNA was used for typing using sequence analysis/PCR with some type-specific primers for bovids, caprids and cervids. The results of the sequence analysis suggested one new putative PV type with sequence identity ranging from 46.45 to 80.09% to other known papillomaviruses, including Capra hircus papillomavirus (ChPV-2), bovine papillomavirus (BPV) 6, 7, 10, 11 and 12, Rangifer tarandus papillomavirus 3 (RtPV-3) and BPV-7Z (Alpine wild ruminant papillomavirus; Cervus elaphus papillomavirus). We therefore propose that this is the first identification of a new putative type, MG523274 (HTY-goat-TR2016), in a goat with teat papillomatosis. It is essential to identify PV types in different animal species and investigate their prevalence/distribution and clinical consequences in order to develop appropriate prophylactic and/or therapeutic procedures and to determine the interspecies transmission potential and evolution of PVs.