Salmonella Typhimurium’da bi̇yofi̇lm oluşumu i̇çi̇n yeni̇ bi̇r regülatör gen: marT

In this study 14 different genes (fimA, fimD, fimF, fimH, stjB, stjC, csgA, csgD, ompC, sthB, sthE, rmbA, fliZ ve yaiC) thought to be related to marT gene at Salmonella enterica serotype Typhimurium 14028 and each involved in the biofilm formation and besides this, MZ1627, a derivative of the same strain that is disrupted in terms of marT gene was examined according to its regulation characteristics with marT. For the comparative analysis of the target genes, each of the gene was mutated through antibiotic gene cassette insertion separately, as repeating the same process to test the autoregulation function of marT gene, 15 different mutant genes in total were obtained. Then, each of these strain's expression level was determined in the presence and absence of the marT induction by transforming recombinant plasmid (pBAD24+marT) into these strains where marT gene's expression is controlled by cloning it into the arabinose-induced BAD promoter. According to QRT-PCR results, marT gene has an important role in terms of regulation of the examined 14 genes. Again these results indicated that, the protein encoded by marT gene is at the same time an autoregulator which organize its own promoter positively, also in general, it suggested that, MarT protein not only regulates the expression of the misL gene encoding an autotransporter protein, but also acts as a global regulator in Salmonella. In this study, strains carrying mutant genes subjected to MarT regulation, compared to parental strain 14028, have also been investigated for their biofilm-forming capacity on polystyrene surfaces. As a result of 24 and 48 hours of incubation, it was observed that there was a statistically significant (p <0.05) decrease in the biofilm formation capacities of each mutant strain. It has been found that fliZ, ompC, rmbA, stjB and stjC genes are directly related to biofilm formation in Salmonella and that there is a serious decrease in biofilm formation in their absence. In the next step, the attachment properties of the mutant strains to the cell surface compared to the parental strain were examined and Caco-2 and HEp-2 cells were used for this. According to the results, single gene mutations are not sufficient for an effective attachment. One of the most important findings of this study is that the global regulator MarT also controls csgD expression, which is the main regulatory gene for biofilm formation. In our "post antibiotics" era caused by the increasing antibiotic resistance crisis, it is thought that the development of chemical inhibitors specific to MarT will play a pioneering role in finding new solutions targeting biofilm formation in Salmonella Typhimurium infections.Bu çalışmada, Salmonella enterica serotip Typhimurium 14028’de marT geni ile ilgili olabileceği düşünülen ve her biri biyofilm oluşumunda rol alan 14 farklı gen (fimA, fimD, fimF, fimH, stjB, stjC, csgA, csgD, ompC, sthB, sthE, rmbA, fliZ ve yaiC) ve bunların yanısıra aynı suşun marT geni bakımından bozulmuş olan bir türevi olan MZ1627 suşu marT ile regülasyon özelliklerine göre incelenmiştir. Adı geçen aday genlerin karşılaştırmalı analizleri için herbir gen antibiyotik kaset mutajenezi (insersiyonu) yoluyla ayrı ayrı mutasyona uğratılmış, aynı işlem, otoregülatör fonksiyonun test edilmesi amacıyla marT geni için de tekrarlanarak toplam 15 farklı mutant suş elde edilmiştir. Daha sonra bu suşların her biri marT geninin arabinoz ile indüklenen BAD promotoru altına klonlanarak ekspresyonunun kontrol altına alındığı rekombinant plasmid (pBAD24+marT) ile transforme edilerek her bir tekli genin ekspresyon seviyesinin marT indüksiyonu varlığında ve yokluğunda nasıl değiştiği belirlenmiştir. QRT-PCR sonuçlarına göre, marT geni, incelenen 14 genin regülasyonu açısından önemli bir role sahiptir. Yine bu sonuçlar, marT geni tarafından kodlanan proteinin aynı zamanda kendi promotorunu da pozitif olarak düzenleyen bir oto-regülatör olduğuna işaret etmiş, genel anlamda da MarT proteininin sadece ototransporter bir protein kodlayan misL gen ekspresyonunu düzenlemekle kalmayıp aynı zamanda Salmonella'da global bir regülatör görevi gördüğünü düşündürmüştür.Bu çalışmada, MarT regülasyonuna tabi tutulan mutant genleri taşıyan suşlar, parental suş 14028’e kıyasla, polistiren yüzeylerde biyofilm oluşturma kapasiteleri de incelenmiştir. 24 ve 48 saatlik inkübasyonlar sonucunda, her bir mutant suşa ait biyofilm oluşum kapasitelerinde istatistiksel açıdan (p< 0.05) dikkate değer bir düşüş olduğu gözlenmiştir. fliZ, ompC, rmbA, stjB ve stjC genlerinin Salmonella’da biyofilm oluşumuyla doğrudan ilişkili oldukları ve yokluklarında biyofilm oluşumunda ciddi bir düşüş olduğu anlaşılmıştır. Bir sonraki aşamada mutant suşların parental suşa kıyasla hücre yüzeylerine tutunma özellikleri incelenmiş ve bunun için Caco-2 ve HEp-2 hücreleri kullanılmıştır. Elde edilen sonuçlara gore, tutunmanın etkili bir şekilde gerçekleşmesi için tek gen mutasyonları yeterli olmamaktadır. Global regülatör MarT’nin, biyofilm oluşumu için başlıca düzenleyici gen olan csgD ekspresyonunu da kontrol etmesi, bu çalışmanın en önemli bulgularından birisidir. Giderek artan antibiyotik dirençliliği krizinin neden olduğu “antibiyotikler sonrası” çağımızda, MarT’ye spesifik kimyasal inhibitörlerin geliştirilmesiyle Salmonella Typhimurium enfeksiyonları özelinde biyofilm oluşumunu hedef alan yeni çözümler bulunması yönünde öncü bir rol oynayacağı düşünülmektedir.Ph.D. - Doctoral Progra

FEN BİLİMLERİ ENSTİTÜSÜ/LİSANSÜSTÜ TEZ PROJESİ

BORDETELLA PERDUSSİS'E AİT REKOMBİNANT PUTATİF PEPTİDOGLİKAN BAĞLAYICI PROTEİN VE ŞAPERONİN 10 (HPS 10) PROTEİNLERİNİN İMMÜN KORUYUCU KAPASİTELERİNİN ARAŞTIRILMAS

FEN BİLİMLERİ ENSTİTÜSÜ/LİSANSÜSTÜ TEZ PROJESİ

BORTELELLA PERTUSSİS'E AİT REKOMBİNANT PUTATİF PEPTİDOGLİKAN BAĞLAYICI PROTEİN VE ŞAPERONİN 10 (Hsp 10) PROTEİNLERİNİN IMMÜN KORUYUCU KAPASİTELERİNİN ARAŞTIRILMAS

FEN BİLİMLERİ ENSTİTÜSÜ/LİSANSÜSTÜ TEZ PROJESİ

BORDETELLA PERTUSSİS'E AİT REKOMBİNANT PUTATİF PEPTİDOGLİKAN BAĞLAYICI PROTEİN VE ŞAPERONİN 10 (HSP 10) PROTEİNLERİNİN IMMÜN KORUYUCU KAPASİTELERİNİN ARAŞTIRILMAS

The education based on toys improved lifelong learning skills oriented effect on students' visual perception skills

1st International Conference on Lifelong Education and Leadership for All -- OCT 29-31, 2015 -- Palacky Univ, Olomouc, CZECH REPUBLICWOS: 000382504900021The research was made for determine the effect of education which consist toys designed lifelong learning skills oriented on 66 month aged students' visual scope abilities. According to this aim, the toys were used which prepared by specialists because of developing the lifelong learning skills. The research was composed of pre-test and post-test semi empirical model and its' sample composed of 104 first class of 66 month aged student including 52 experimental group and 52 control group which were training in Duzce city primary schools depended On National Education Ministry Before starting the education the test entitled 'Frostig Developmental Test Of Visual Perception' was developed by Marianne Frostig and studied of validity and reliability by Sokmen (1994) was used as pre-test and post-test. For education programme, the ambiance was disagned for purpose by specialists and trained during the 12 week of 2 days 60 min. per a week. The datas, according the research results were determined by SPSS packet programme and the education orientation of toys based on lifelong learning skills was stated that the education differentiated the 66 month aged of students' visual memory and percept mechanisms for positive rounded significantly.Moravian Univ Coll Olomou

Regulation of biofilm formation bymarTinSalmonellaTyphimurium

In this study, we aimed at identifying the regulatory role ofmarTgene, known as the regulator ofmisL, on 15 different biofilm-related genes inS.Typhimurium 14028 strain. We also tested the strains for their ability to form biofilm and determined the adherence characteristics of the wild type and the mutant strains of the organism on Caco-2 and HEp-2 cells. For comparative analyses of the candidate genes, individual gene mutations were created via antibiotic gene cassette insertion into each gene of interest.marTgene was cloned behind an arabinose inducible BAD promoter in order to controlmarTexpression. This recombinant plasmid was transfer into each of the 15 mutant strains to investigate the level of expression of each single gene in the presence and absence ofmarTinduction. Besides determination of variations in biofilm formation by each mutant strain, the attachment characteristics of them onto Caco-2 and HEp-2 cell lines were also reported. As a result of attachments experiments on polystyrene surfaces, it was determined that the biofilm production capacity of each mutant strain decreased in a statistically significant manner (p < 0.05). QRT-PCR trials indicated that themarTgene regulates the expression of 14 genes, namelyfimA,fimD,fimF,fimH,stjB,stjC,csgA,csgD,ompC,sthB,sthE,rmbA,fliZandyaiC, in a positive manner. QRT-PCR studies were also revealed that the MarT protein positively regulates its own promoter. When the adherence characteristics of the mutant strains and the wild-type were investigated by using Caco-2 and HEp-2 cells, it was determined that the single gene mutations did have no effect on bacterial adhesion. In view of our mutational analyses and biofilm formation studies, it was concluded thatfliZ,ompC,rmbA,stjBandstjCgenes are related with biofilm formation inSalmonella,besides other cellular functions of them. Taken together, our data suggested that the regulatory role of MarT protein is not only restricted to the regulation ofmisLgene expression, but it rather acts as a general regulator on the biofilm-related genes inSalmonella

The Preschool Teachers’ Attitudes About Education of Multiculturalism

Multicultural education; ethnicity, religion, race, language, gender, where cultural properties due to differences such as the living area, to reduce identity conflicts and power sooner quinidine, society, philosophy of education that appeal to different cultural characteristics and be reflected in the formal education of the application and based on democratic values, to promote cultural diversity equality training which is defined as a mixed school climate.This study was conducted to determine the pre-school teachers in multiculturalism and cognitive levels of knowledge and attitudes of multicultural education. The study population consisted of teachers in pre-school located in Düzce. The sample of the study in Düzce province in formal kindergarten and kindergarten assistants, including 5 men and 86 women constitute 91 pre-school teachers. Joseph research within the framework of quantitative research methods Ponterotto G. (1998) led a group of researchers developed by the Multicultural Education Teacher Attitudes (OCTO) is used. Research has established formal working group of kindergarten and pre-school teachers working in kindergarten. The study was carried out with the teacher. The scale used in this study, Cronbach alpha reliability coefficient of .74 bulunarak as is provided for translation and language validity. Factor analysis showed that the scale of the one-dimensionality of supported theoretically predicted. Age according to the result, graduated region where the school is located, where the school their seniority and job place and according to the cultural structure of the occurrence of settlement of prior training for the various variables was examined whether there is a difference. As a result, the location of the school they graduated teachers, seniority, school, depending on whether they work in the province or district was found that differences in the approach to multicultural education

Ribavirin shows antiviral activity against SARS-CoV-2 and downregulates the activity of TMPRSS2 and the expression of ACE2 In Vitro

Ribavirin is a guanosine analog and has a broad-spectrum antiviral activity against RNA viruses. Based on this, we aimed to show the anti-SARS-CoV-2 activity of this drug molecule via in vitro, in silico and molecular techniques. Ribavirin showed antiviral activity in Vero E6 cells following SARS-CoV-2 infection whereas the drug itself did not show any toxic effect over the concentration range tested. In silico analysis suggested that Ribarivin has a broad-spectrum impact on SARS-CoV-2, acting at different viral proteins. According to the detailed molecular techniques, Ribavirin was shown to decrease the expression of TMPRSS2 both at mRNA and protein levels 48 hours after treatment. The suppressive effect of Ribavirin in ACE2 protein expression was shown to be dependent on cell types. Finally, proteolytic activity assays showed that Ribavirin also showed an inhibitory effect on TMPRSS2 enzyme. Based on these results, we hypothesized that Ribavirin may inhibit the expression of TMPRSS2 by modulating the formation of inhibitory G-quadruplex structures at the TMPRSS2 promoter. As a conclusion, Ribavirin is a potential antiviral drug for the treatment against SARS-CoV-2, and it interferes with the effect of TMPRSS2 and ACE2 expression.The accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author