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Öğe Applicability evaluation of mtDNA based molecular identification in mosquito species/subspecies/biotypes collected from Thessaloniki, Greece(Elsevier B.V., 2023) Köseoğlu, A.E.; Paltacı, S.; Can, H.; Giantsis, I.A.; Güvendi, M.; Demir, S.; Döşkaya, M.The genus Culex, containing many described species, plays a role as a vector for diseases of medical and veterinary importance worldwide. Among these species, Culex pipiens is one of the most widespread mosquitoes and is classified into two biological forms (biotypes), named as Culex pipiens pipiens and Culex pipiens molestus. Due to similar morphological structure between these biotypes, morphological identification is inadequate. Thus, molecular methods have been developed and are considered more reliable, some of which are based on analyses of mitochondrial DNA. The aim of the present study was to evaluate the applicability and reliability of mtDNA based molecular identification methodologies. Initially, mosquito specimens (n = 100) collected from Thessaloniki, Greece were morphologically analyzed. Then, mitochondrial cox1 sequencing and PCR-RFLP methods were used to confirm the morphological identification results as well as to discriminate species and subspecies/biotype of Culex pipiens complex. According to morphological identification results, Culex pipiens complex (n = 92), Culex modestus (n = 6) and Culex theileri (n = 2) were detected. Using mtDNA sequencing, all Culex modestus and Culex theileri samples were confirmed whereas 86 of Culex pipiens complex were detected as Culex pipiens but surprisingly the remaining six of them were detected as Culex quinquefasciatus. Among Culex pipiens specimens, PCR-RFLP detected that frequency of Culex pipiens pipiens (85%; 85/100) was very high compared to Culex pipiens molestus (1%, 1/100). In conclusion, this study shows the necessity of use of molecular methods beside morphological methods for especially specimens detected as Culex pipiens. Also, it was shown that mtDNA PCR-RFLP methodology represents a well-established alternative for Culex biotypes identification. © 2023 Elsevier B.V.Öğe CHARACTERISTICS OF VARIOUS CELL TYPES IN PRIMARY INSECT CELL CULTURE FROM LARVAL OVARIES OF SILKWORM (BOMBYX MORI L.)(Pakistan Agricultural Scientists Forum, 2019) Soya, S.; Can, H.As insect cell lines can be used in baculovirus expression systems for producing recombinant proteins, there is a need to establish new cell lines. Insect cell lines are thought to be useful in production of humanized recombinant proteins. Hence, we aimed to establish a culture from larval ovaries of Bombyx mori to investigate the cell characteristics and growth properties. Two sets of primary cell cultures were established from ovaries of fifth instar larvae. Characterization of cell line was carried out by comparing the PCR products of cell line and silkworms from COI (cytochrome c oxidase subunit I) specific primers. In the first set of cultures, the culture population was found to be heterogeneous and cell sizes were variable. Fibroblastic, epithelial-like, spindle-like, and round cells were observed after 7th day. Interestingly, contractions were observed in muscle-like cells at the end of first month, and stayed stable until the fourth month. At the beginning of the fourth month, because cell death increased significantly, cell concentrations decreased. In the second set of cultures, it was determined that doubling time was about 72 h with high viability and confluency of cells were higher in early passages. PCR which was conducted for characterization of cells resulted in a 710 bp product in each cell line and silkworms. In conclusion, growth rate detected in this study was observed to be compatible with literature. Also, it was thought that if primary culture is supported with growth stimulators, growth rate can be more accelerated and this primary culture can be used more efficiently in recombinant protein production studies.Öğe Cryptosporidiosis outbreak on a dairy farm: Detection of Cryptosporidium parvum as a causative agent in the water source(Polska Akad Nauk, Polish Acad Sciences, Univ Warmia & Mazury Olsztyn, 2021) Karakavuk, M.; Can, H.; Doskaya, M.; Karakavuk, T.; Erkunt-Alak, S.; Koseoglu, A. E.; Gul, A.Diarrhea caused by parasitic agents is common in neonatal calves and leads to significant economic losses in cattle farms worldwide. Cryptosporidium spp. is one of the most frequently detected parasitic agents causing diarrhea in neonatal calves. The aim of this study was to investigate the presence of Cryptosporidium spp. on a dairy farm which a has major diarrhea problem. Samples were collected from calves, cows, drinking bowls, and two different artesian water sources, as well as from the environment. All fecal samples were investigated using Kinyoun acid-fast stained slides and real-time PCR targeting the Cryptosporidium spp. COWP gene. In addition, species identification was performed by nested PCR targeting the Cryptosporidium spp. COWP gene and sequencing. Cryptosporidium spp. was detected in 11 calves (30.55%; 11/36) by real-time PCR and the cows were negative. Among real-time PCR positive samples, only five were also found positive by microscopy. Moreover, Cryptosporidium spp. was found in one of the two artesian water sources and five environmental samples by real-time PCR. Among these positive samples, eight were sequenced. According to the RFLP pattern, BLAST and, phylogenetic analyses, all sequenced samples were Cryptosporidium parvum. These findings show the importance of C. parvum as a cause of calf diarrhea on dairy farms.Öğe Development and Sensitivity Determination of 18S rRNA Gene-specific Fast Loop-mediated Isothermal Amplification (LAMP) Assay for Rapid Detection of Acanthamoeba(Galenos Publishing House, 2023) Aykur, M.; Karakavuk, M.; Akıl, M.; Can, H.; Döşkaya, M.; Gürüz, A.Y.; Dağcı, H.Objective: Acanthamoeba, one of the free-living amoeba, has been detected in many environmental samples, mainly in water, soil and air. Acanthamoeba keratitis and granulomatous amoebic encephalitis are among the most important clinical manifestations caused by Acanthamoeba. In this study, it was aimed to determine the sensitivity of the rapid loop mediated isothermal amplification (LAMP) test designed with primers specific to Acanthamoeba 18S rRNA gene to detect more rapidly the presence of Acanthamoeba in clinical and environmental samples. Methods: Acanthamoeba strain grown in culture was diluted in 200 µL as 1x106 trophozoites and DNA was isolated, and the amount of DNA was determined by Nano-Drop Spectrophotometer. The purified DNAs were diluted from 1000 pg to 0.001 pg and used in colorimetric and fluorescence-based LAMP reactions. The LAMP reaction mixture was incubated for 60 minutes at 63 °C in a total volume of 25 µL. Results: To determine the sensitivity of the test, positivity of Acanthamoeba genomic DNA was observed at 1, 10, 100 and 1000 pg/reaction in both colorimetric and fluorescence-based LAMP tests. The lowest analytical sensitivity of both calorimetric and fluorescent LAMP assay was determined as 1 pg/ reaction. In addition, as a result of LAMP reaction applied with other parasite DNAs to evaluate the specificity of the test, no LAMP product was detected in calorimetric and 1% agarose gel electrophoresis, except for positive control, and the specificity of the test was determined as 100%. Conclusion: It has been demonstrated that the LAMP assay designed by targeting 18S rRNA gene of Acanthamoeba has a detection limit of 1 pg of genomic DNA. It is promising that LAMP test is more sensitive and faster than culture method, as well as simple, inexpensive and highly sensitive. For this reason, it is thought that developed test can be applied in the diagnosis of Acanthamoeba in environmental and clinical samples. © 2023, Galenos Publishing House. All rights reserved.Öğe Genetic characterization of Bartonella henselae samples isolated from stray cats by multi-locus sequence typing(BioMed Central Ltd, 2023) Can, H.; Güvendi, M.; Sürgeç, E.; Köseoğlu, A.E.; Erkunt, Alak, S.; Karakavuk, M.; Gül, A.Background: Bartonella henselae is one of the most commonly identified Bartonella species associated with several human diseases. Although B. henselae was detected in humans and cats in Turkey, they have not been genotyped previously. Therefore, this study aimed to genotype B. henselae samples (n = 44) isolated from stray cats using the multi-locus sequence typing (MLST) method. For this aim, eight different housekeeping markers were amplified by nested PCR and then sequenced to reveal sequence types (STs) of B. henselae samples. Results: Allelic profiles obtained from 40 B. henselae isolates (90.9%) were compatible with available allelic profiles in the MLST online database. However, allelic profiles obtained from the remaining 4 B. henselae isolates (9.1%) were incompatible with the database. Among B. henselae isolates with compatible allelic profiles, 5 different STs including ST1, ST5, ST9, ST35 and ST36 were identified according to the B. henselae MLST online database. ST35 was the most prevalent ST with a prevalence rate of 29.5% (13/44), followed by ST36 with a prevalence rate of 22.7% (10/44). In addition, ST5 (16%, 7/44) and ST9 (18.2%, 8/44) were also among the prevalent STs. The prevalence of ST1 was 4.5% (2/44). For B. henselae isolates with incompatible allelic profiles, we recommended a new ST called ST38. Conclusion: The present study genotyped B. henselae samples isolated from stray cats in Turkey for the first time and ST1, ST5, ST9, ST35, and ST36 as well as a new sequence type named ST38 were identified among these B. henselae isolates. © 2023, BioMed Central Ltd., part of Springer Nature.Öğe Importance of screening severe COVID-19 patients for IFN-?1, IL-6 and anti-S1 IgG levels(Academic Press, 2023) Kenanoğlu, O.B.; Gül, A.; Can, H.; Karakavuk, M.; Erkunt, Alak, S.; Korukluoğlu, G.; Altaş, A.B.Cytokine storm is an important cause of death in COVID-19 patients. A recent clinical study showed that administration of recombinant interferon lambda 1 (IFN-?1 or IL-29) may prevent severe COVID-19. On the other hand, IL-6 has been associated as a prognostic marker of worsening for COVID-19 patients. The objective of this study is to screen IFN-?1, IL-6 and antibody levels in consecutive serum sample sets of COVID-19 patients. A total of 365 serum samples collected from 208 hospitalized COVID-19 patients were analyzed for IFN-?1 and IL-6 levels as well as SARS-CoV-2 neutralizing antibodies and anti-S1 IgG antibodies. Analyses of serum samples for cytokine levels showed that IFN-?1 (>8 pg/mL) and IL-6 (>2 pg/mL) were detected in approximately 64% and 21% patients, respectively. A decrement in IFN-?1 levels and IL-6 levels above 35 pg/mL can be sign of clinical severity and upcoming dead. An increment in IL-6 levels wasn't detected in every COVID-19 patient but a decrement in IL-6 levels was related to clinical improvement. Importantly, the detection of IFN-?1 level together with an increase in anti-S1 IgG antibody response were observed in clinically improved patients. Screening severe COVID-19 patients for IFN-?1, IL-6, and anti-S1 IgG antibody levels during their hospital stay especially in intensive care units may be beneficial to monitor the clinical status and management of treatment strategies. Importantly, detection of IFN-?1 together with protective IgG antibody response can be an indication of clinical improvement in severe COVID-19 patients and these patients may be discharged from the hospital soon. © 2023 Elsevier LtdÖğe The Importance of Vaccines in a Sustainable Healthy Society(Springer International Publishing, 2023) Gül, A.; Alak, S.E.; Can, H.; Ün, C.; Gül, C.; Karakavuk, T.B.; Karakavuk, M.A sustainable society is a community that can live within the frame of the ecosystem. If the actions that put excessive burden on the ecosystem can be resolved, sustainable society can be preserved as a collective and ongoing entity. New o utbreaks are emerging at an unprecedented rate, causing serious damage to sustainable society. Pandemic outbreaks have often emerged in an unpredictable manner and are a major obstacle to a sustainable society because of increase in use of non- recyclable waste and disposable goods. In addition, increase in unemployment, insufficient education, and decreased interpersonal communication during the outbreaks negatively impact sustainable society. Vaccines have the power to support sustainable development goals by creating herd immunity in societies, which reduces the incidence of disease, prevents transmission, and reduces the social and economic burden of the disease. High vaccination coverage can effectively combat the outbreaks, prevent secondary infection waves, and control seasonal endemic outbreaks of infection which in turn supports sustainable society. This effect of vaccines has been showed in history by eliminating diseases like smallpox and polio and combatting all other vaccine-preventable diseases. In this section, we highlighted the importance of the vaccines for a sustainable society in terms of health perspectives. © The Editor(s) (if applicable) and The Author(s), under exclusive license to Springer Nature Switzerland AG 2023.Öğe An In house ELISA using recombinant GRA1 and BAG1 proteins as antigen successfully detected Toxoplasma gondii infection in a flock of sheep suffering from abortions(Elsevier B.V., 2023) Kandemir, Ç.; Can, H.; Karakavuk, M.; Döşkaya, M.; Erkunt, Alak, S.; Gürüz, A.Y.; Ün, C.Toxoplasma gondii is one of the main pathogens causing abortion in sheep. In this study, an in house ELISA using recombinant GRA1 and BAG1 proteins as antigen was used to detect T. gondii infection in a flock of sheep suffering from abortions. Serum samples collected from sheep (n = 23) the day after abortion were initially analyzed with a commercial ELISA using recombinant P30 (SAG1) protein as antigen and then by the in house ELISA using recombinant GRA1 and BAG1 proteins as antigen. Commercial ELISA detected anti-T. gondii IgG antibodies in 20 samples whereas in house ELISA detected anti-T. gondii IgG antibodies in 19 samples. The results of the remaining three serum samples that were seronegative by commercial ELISA were also seronegative by in house ELISA. According to the results of commercial ELISA and in house ELISA, the seroprevalence of ovine toxoplasmosis was 86.9% and 82.6%, respectively. When we accept the validated commercial ELISA as a reference method, the sensitivity and specificity of in house ELISA can be considered as 95% and 100%. These preliminary results show that in house ELISA using recombinant GRA1 and BAG1 proteins as antigen can detect ovine toxoplasmosis successfully. © 2023Öğe In silico discovery of epitopes of gag and env proteins for the development of a multi-epitope vaccine candidate against Maedi Visna Virus using reverse vaccinology approach(Academic Press, 2023) Koçkaya, E.S.; Can, H.; Yaman, Y.; Ün, C.Maedi Visna Virus (MVV) causes a chronic viral disease in sheep. Since there is no specific therapeutic drug that targets MVV, development of a vaccine against the MVV is inevitable. This study aimed to analyze the gag and env proteins as vaccine candidate proteins and to identify epitopes in these proteins. In addition, it was aimed to construct a multi-epitope vaccine candidate. According to the obtained results, the gag protein was detected to be more conserved and had a higher antigenicity value. Also, the number of alpha helix in the secondary structure was higher and transmembrane helices were not detected. Although many B cell and MHC-I/II epitopes were predicted, only 19 of them were detected to have the properties of antigenic, non-allergenic, non-toxic, soluble, and non-hemolytic. Of these epitopes, five were remarkable due to having the highest antigenicity value. However, the final multi-epitope vaccine was constructed with 19 epitopes. A strong affinity was shown between the final multi-epitope vaccine and TLR-2/4. In conclusion, the gag protein was a better antigen. However, both proteins had epitopes with high antigenicity value. Also, the final multi-epitope vaccine construct had a potential to be used as a peptide vaccine due to its immuno-informatics results. © 2023 International Alliance for Biological StandardizationÖğe Mitotyping of random bred cats and pure breed cats (Turkish Angora and Turkish Van) using non-repetitive mitochondrial DNA control region(Elsevier B.V., 2024) Bilgen, N.; Güvendi, M.; Sezgin, E.; Can, H.; Köseoğlu, A.E.; Erkunt, Alak, S.; Döşkaya, M.The Fertile Crescent appears to be the most plausible region where the domestication of cats commenced through a mutually beneficial relationship between wild cats and early agrarian societies. These domesticated cats then journeyed across the globe mirroring the paths of human migration. An examination of mitochondrial DNA (mtDNA) control region-based mitotyping suggested that a significant majority, exceeding 80%, of globally sampled random-bred and pure-bred cats could be categorized into 12 predominant mitotypes. However, the extent of mitotype diversity within random-bred cats from regions proximate to the Fertile Crescent remains inadequately explored. In light of this we aimed to investigate the mitotype diversity in random bred cats sampled from various regions across Turkey. Additionally, we sought to establish a comparison with the mitotype profiles of locally recognized pure breeds, namely the Turkish Angora and Turkish Van. To unravel their evolutionary narratives, we engaged in comprehensive population genetics analyses at both the individual and mitotype-based levels. Our study encompassed a sample size of 240 specimens, forming the basis for both mitotyping and population genetics scrutiny. Our analysis yielded the identification of nine ‘universal’ mitotypes (A—J), alongside an ‘outlier’ mitotype group I. Notably mitotypes A and D emerged as particularly prevalent in contrast to the lesser occurrence mitotypes C, G, and H. With the realm of random bred cats the structure of haplotypes exhibited remarkable diversity presenting distinctions from Turkish Angora and Van breeds. Nucleotide diversity was higher compared to previous reports from Turkey and was one of the highest among reported world cat population estimates. Intriguingly, our investigations did not unveil any pronounced instances of strong selection, population expansions or contractions within any specific population or mitotype. To conclude, our study represents a pioneering effort in uncovering the mitotype profiles and haplotype structures inherent to both random-bred and pure breed cats in Turkey. This endeavor not only broadens our understanding of the feline genetic landscape within the region but also lays the foundation for future inquiries into the evolutionary trajectories and genetic legacies of these feline populations. © 2023 Elsevier B.V.Öğe Molecular and bioinformatics evaluation of PRNP as a candidate gene for successful suicidal behavior(Elsevier Masson s.r.l., 2023) Cun, E.; Can, H.; Köseoğlu, A.E.; Erkunt, Alak, S.; Koyuncu, G.; Ün, C.Contemplating suicide, suicide attempts, and successful suicides represent the suicidal behavior accompanied by depression and hopelessness. Suicide is among the major causes of death worldwide. According to obtained data, it has been reported that approximately 800,000 suicide deaths occur worldwide every year and the frequency of suicide varies from country to country. Many suicide attempts or the contemplation of suicide are associated with psychological disorders such as depression and schizophrenia as well as alcoholism and drug abuse. It has been demonstrated that depression, which affects nearly 350 million people worldwide is strongly linked to suicidal behavior, as it is the most common disorder in people who die from suicide. There are several major hypotheses in the pathogenesis of depression. One of them is the monoamine hypothesis and the insufficiency of monoamine neuromediators such as serotonin, norepinephrine, and dopamine in the central nervous system is considered to be a triggering factor for the development of depression. Another hypothesis is the stress hypothesis, and chronic stress and stressful life events are accepted as a predictor in the onset of depression. The third one is the disturbance of neurogenesis and neuroplasticity caused by especially brain-derived neurotrophic factors (BDNF) in nervous tissue. Another depression development related theory is the cytokine theory explained by the activation of the inflammatory response system inducing the various behavioral, neuroendocrine, and neurochemical alterations. Finally, the circadian rhythm theory that is responsible for regulating many physiological and behavioral processes and sleep disorders are reported to occur in most of the individuals suffering from depression. The findings about physiological roles or normal functions of the prion protein are very limited but there are several studies showing the possible role of prion protein in circadian rhythm, sleep deprivation and development of depression using mice devoid of prion protein. Relying on these findings, we sequenced the prion protein coding genes in a cohort of individuals who had successfully committed suicide (n = 75) and compared allele and genotype frequencies with a control group (n = 44). We also applied bioinformatics analysis to detect the interaction between the prion protein Met/Val allele and DRD2, PER2, PER3, and COMT proteins, which are very present in the brain and related to depression, stress, and circadian rhythm. Among the individuals who died by suicide, although the frequency of the homozygous Met/Met genotype (45/75; 60%) was higher compared to the control group (20/44; 45.45%), no statistically significant difference was found (X2 = 2.530; P = 0.282). Parallel to this, the frequency (76.6%) of Met allele detected in the individuals who died by suicide was also higher than that of (67%) the control group, but no statistically significant difference was found (X2 = 4.338; P = 0.114). Docking analysis also showed that the prion protein with Val allele had a higher binding affinity with proteins analyzed according to the Met allele. In conclusion, the Met/Met genotype and Met allele frequencies were detected to be higher in the completed suicidal group. Also, the interaction between the prion protein with the M allele and proteins analyzed was lower. Based on these findings, it was thought that carriers of the prion Met allele may be more prone to suicide. © 2023 Elsevier Masson SASÖğe Molecular prevalence and subtyping of Blastocystis sp. isolates in stray cats of Izmir, Turkey: First report of ST4 allele 42 in cats(Polska Akad Nauk, Polish Acad Sciences, Univ Warmia & Mazury Olsztyn, 2021) Can, H.; Koseoglu, A. E.; Alak, S. Erkunt; Guvendi, M.; Un, C.; Karakavuk, M.; Doskaya, A. DegirmenciBlastocystis sp. is one of the most frequently detected intestinal parasites in humans and can inhabit a wide range of animals. Close contact with animals is one of the transmission factors of Blastocystis sp. infection in humans. In this study, we aimed to investigate the molecular prevalence and subtypes of Blastocystis sp. in stray cats living in Izmir, Turkey. The PCR targeting the barcode region in the SSU rRNA gene was performed with DNA samples isolated from feces (n:465) to investigate the presence of Blastocystis sp. PCR positive samples were sequenced for subtyping analysis. Among the samples analyzed, Blastocystis sp. DNA was detected in 17 (3.65%) of them and sequence data were obtained from only seven isolates. Phylogenetic analysis showed that seven Blastocystis sp. isolates clustered with the reference Blastocystis ST4 isolates. Similarity rates were between 83.22% and 99.25%. In addition, Blastocystis database results confinned that all of these were allele 42 corresponding to ST4. As a result, the present study shows for the first time the presence of ST4 allele 42, the prevalent subtype in humans, in stray cats in Izmir, Turkey. This finding supports the notion that stray cats can be a source of Blastocystis sp. infection in humans.Öğe Molecular prevalence of Enterocytozoon bieneusi in stray cats of İzmir, Türkiye(Elsevier Ltd, 2023) Erkunt, Alak, S.; Can, H.; Değirmenci, Döşkaya, A.; Sürgeç, E.; Güvendi, M.; Ün, C.; Döskaya, M.The phylum Microsporidia contains obligate single celled parasites that can infect many vertebrate hosts including humans. Enterocytozoon bieneusi is considered as the most diagnosed species in humans. E. bieneusi has also been detected in many animals such as cats, dogs and cattle. Among these animals, cats are carriers of type D and IV which are the most common human pathogenic genotypes of E. bieneusi. In Türkiye, the prevalence of E. bieneusi in stray cats is not well known. Therefore, in this study, the molecular prevalence of E. bieneusi in stray cats (n = 339) was determined by Real-Time PCR targeting ribosomal DNA ITS (internal transcribed spacer) region of E. bieneusi. Initially, the analytical sensitivity of Real-Time PCR was determined by a plasmid control and then E. bieneusi DNA was investigated in fecal samples of stray cats. The results showed that the analytical sensitivity of Real-Time PCR targeting ITS region of E. bieneusi was ?1 copy plasmid/reaction. Analysis of fecal samples revealed that the molecular prevalence of E. bieneusi was 50.15% (170/339). Overall, these results showed that the Real-Time PCR successfully detected E. bieneusi in cat's fecal samples and stray cats can be an important source for transmission of E. bieneusi to humans and other animals. © 2023 Elsevier LtdÖğe A preliminary study to develop a lateral flow assay using recombinant GRA1 protein for the diagnosis of toxoplasmosis in stray cats(Elsevier Ltd, 2023) Değirmenci, Döşkaya, A.; Can, H.; Gül, A.; Karakavuk, T.; Güvendi, M.; Karakavuk, M.; Gül, C.Toxoplasma gondii is a protozoan parasite that may infect many mammals including humans. Cats are one of the main sources of infection for humans. Therefore, routine screening of cats with tests that are inexpensive, rapid, and do not require sophisticated laboratory equipment is important. In this study, a lateral flow assay (LFA) was designed to rapidly diagnose toxoplasmosis in cats. For this purpose, we selected GRA1 protein of T. gondii due to its high antigenicity in diagnostic and vaccine studies. We further analyzed the immunological properties of GRA1 protein using in silico tools. Then, we expressed and purified recombinant GRA1 (rGRA1) protein and used it during the development of LFA to detect toxoplasmosis in serum samples (n = 40) of cats. According to the results, rGRA1 protein has negative GRAVY value, high aliphatic index, alpha helix, random coil and 12 B cell epitopes. The in silico data supported the high antigenic properties of rGRA1 protein and showed that it can be a good antigen candidate for LFA. Among 30 cat positive serum samples, 27 were found positive by the LFA while seronegative sera (n = 10) were negative by the LFA. The preliminary data showed that the LFA has high sensitivity (90 %) and specificity (100 %). When we used high responsive cat sera (i.e. sera that have optical density > 0.5 with ELISA) the sensitivity value reached 100 %. These results showed that rGRA1 protein is a good candidate to develop a LFA for rapid diagnosis of toxoplasmosis in cats. © 2023 Elsevier LtdÖğe Toxoplasma gondii destroys Her2/Neu-expressing mammary cancer cells in vitro using a continuous feed medium approach(NLM (Medline), 2020) Atalay Şahar, E.; Döşkaya, M.; Karakavuk, M.; Can, H.; Gül, A.; Gürüz, A.Y.; Yeniay, L.INTRODUCTION: Toxoplasma gondii is an opportunistic protozoan and can be grown using several human cell lines. Breast cancer is the second leading cause of cancer death in women. Her2/Neu-expressing mammary cancer cell lines called TUBO can be grown in vitro. In recent years, protozoan parasites have become popular means of use in cancer therapy research. In this study, we analyzed whether T. gondii tachyzoites can destroy TUBO cells using a novel continuous feed medium approach. METHODOLOGY: Two sets of flasks (each containing four groups) containing TUBO cells were inoculated with T. gondii Ankara strain tachyzoites. First set containing 5×106 TUBO cells were inoculated with TUBO-tachyzoite ratios of 1:2, 1:1, 2:1, and 4:1 and second set containing 1×106 TUBO cells were inoculated with TUBO-tachyzoite ratios of 10:1, 100:1, 1000:1, and 2000:1. Thereafter, culture supernatants were harvested at various days until TUBO cells were destroyed and tachyzoites were counted. RESULTS: In the first and second sets of flasks, TUBO cells were destroyed between days 8 to 12 and 12 to 25, respectively. In addition, the amount of tachyzoites increased 7- 43 and 595 to 112500 times in the first and second set of flasks, respectively. CONCLUSIONS: These results show that T. gondii tachyzoites successfully destroy Her2/Neu-expressing mammary cancer cells using a continuous feed medium approach. Although this idea may be too premature for the moment, the approach defined herein may support future researchers investigating the relationship between cancer and parasites which can make important progress toward saving cancer patient lives. Copyright (c) 2020 Esra Atalay Sahar, Mert Doskaya, Muhammet Karakavuk, Huseyin Can, Aytul Gul, Adnan Yuksel Guruz, Aysu Degirmenci Doskaya, Levent Yeniay.Öğe Tumour forming efficiencies of TUBO cell line expressing Her2 in two different mice strains with Matrigel matrix to develop animal models for novel immunotherapies(Elsevier Sci Ltd, 2016) Anil, M.; Iz, S. Gulce; Metiner, P. Saglam; Sahar, E. Atalay; Can, H.; Zekioglu, O.; Doskaya, M.; Yeniay, L.
