Immobilization of Bacillus Subtilis E6-5 Protease and Commercial Protease in Nanofibrils Containing Different Amino Acids

DergiPark: 633788trkjnatBu çalışmada, elektrospin yöntemiyle yüksek yüzey alanına sahip, glisin, tirozin ve glutamik asit aminoasitleri ile oluşturulmuş poliamid 6 polimer yüzeyler üretilmiş ve liyofilize Bacillus subtilis E6-5 proteaz ve ticari proteaz enzimleri nanofibriller üzerinde immobilize edilmiştir. Enzimlerin yeniden kullanılabilirliği araştırıldı. Enzimlerin immobilizasyon verimlilikleri yaklaşık olarak % 50-55 arasındaydı. Liyofilize Bacillus proteazı ile yapılan çalışmalarda glutaraldehitle aktifleştirilmiş PA6 nanolifler ve glutaraldehitle aktifleştirilmeyen PA6 nanoliflerde glutamik asit aminoasidi varlığında immobilizasyonun daha başarılı olduğu saptanmıştır. Glutaraldehit ile aktifleştirilmemiş ve aktifleştirilmiş yüzeylerde immobilize edilen liyofilize proteaz enziminin 4 kez kullanımı olmasına rağmen, en iyi işlevsel stabilite 2 kez kullanım ile elde edilmiştir. Saf PA6/glutamik asit nanoliflerinde iki tekrarlı kullanım sonucu enzimin immobilizasyon verimi % 38 olarak bulunmuştur. Glutaraldehitle aktifleştirilmiş PA6 nanoliflerde de PA6/glutamik asit nanolif yüzeyleri iki tekrarlı kullanım sonucu enzimin immobilizasyon verimi % 65 olarak bulunmuştur. Nanoliflerin glutaraldehitle aktifleştirmesi sonucu enzim immobilizasyon verimi iki kat arttırılmıştır. Ticari proteaz ile yapılan çalışmalarda ise glutaraldehitle aktifleştirilmemiş nanolif yüzeylerde enzimin 6 kez kullanımı olmasına rağmen en işlevsel stabilite 3 tekrarlı kullanımda elde edilmiştir. En başarılı immobilizasyon verimi PA6 nanoliflerde % 58 olarak bulunmuştur. Glutaraldehitle aktifleştirilmiş PA6 nanoliflerde de enzim 6 kez kullanım bulmuş fakat işlevsel stabilite 4 tekrarlı kullanıma kadar korunmuştur.In this study, polyamide 6 polymer surfaces that have a high surface area were produced by electrospinning method with the participation of Glycine, Tyrosine and Glutamic acid amino acids, and lyophilized Bacillus subtilis E6-5 protease and commercial protease enzymes were immobilized on nanofibrils. Enzyme reusability were investigated. The immobilization efficiencies of the enzymes were approximately between 50-55 %. In studies with lyophilized Bacillus protease, glutaraldehyde activated PA6 nanofibrils and glutaraldehyde unactivated PA6 nanofibrils were found to be more immobilized in the presence of Glutamic acid. Although the lyophilized protease enzyme immobilized on non-glutaraldehyde activated and activated surfaces has been used 4 times, the best functional stability has been achieved with 2 times use. In pure PA6/Glutamic acid nanofibrils, the immobilization yield of the two times used enzymes was found to be 38 %. In glutaraldehyde-activated PA6 nanofibrils, the PA6/Glutamic acid nanofibril surfaces were found to have 65 % immobilization yield of the two repetitive used enzymes. The enzyme immobilization efficiency has been doubled by glutaraldehyde activation of the nanofibrils. In studies with commercial protease, the most functional stability was obtained for 3 repeated uses, although the enzyme was used 6 times on the non-glutaraldehyde activated nanofibril surfaces. The most successful immobilization was found in 58 % of PA6 nanofibrils. In glutaraldehyde-activated PA6 nanofibrils, the enzyme was found to be used 6 times, but the functional stability was maintained as much as 4 times of repeated use

Serum Ischaemia-modified albumin concentration in hyperemesis gravidarum

Introduction: The role of oxidative stress in the pathogenesis of hyperemesis gravidarum (HEG) has been demonstrated in a lot of studies. The present study aimed to compare the ischaemia-modified albumin (IMA) serum levels of patients diagnosed with HEG with healthy pregnant women, and to investigate whether oxidative stress caused increased serum IMA in HEG. Methods: Pregnant women were classified into a group diagnosed with HEG (n=45) and an age- and body mass index-matched control group without HEG (n=45). Serum IMA levels were assessed by the enzyme-linked immunosorbent assay (ELISA) method. Results: Serum IMA levels were higher in the HEG group than the control group (HEG: 8.2 +/- 0.2 ng/mL, control: 6.9 +/- 03 ng/mL, p<0.001). Conclusion: We found that HEG was related to increased maternal serum IMA levels. The high levels of IMA in HEG can he considered as a reflection of increased oxidative stress

Investigation of the effects of mir-219-1 gene variants on the development of disease in non-small cell lung cancer patients

Background: Various variants of the miR-219-1 gene are one of the first genes associated with NSCLC prognosis in the literature. Objectives: We aimed to genotype two different variants of the miR-219-1 gene and to investigate to using of the result as a biomarker in the diagnosis and treatment of NSCLC. Materials and Methods: The patients were chosen according to International NSCLC criteria and genomic DNA was isolated from blood (138 patients and 100 healthy individuals). Then qRT-PCR was applied to determine the rs213210 and rs421446 variants of miR-219-1 gene polymorphisms. Allele and genotype frequencies were compared using Pearson’s chi-square and Fisher’s exact tests test. Results: We found that TT genotype (p=0,381) in rs213210 compared with CC genotype (p=0,165) and CC genotype (p=0,823) in rs421446 compared with TT genotype (p=0,537) did not show a significantly increased risk of NSCLC. There is no relationship between polymorphisms in miR-219-1 and the outcome of NSCLC. Conclusion: miRNA single nucleotide polymorphisms can be used as genetic biomarkers to predict cancer susceptibility, early diagnosis, and prognosis. Our study has shown that two variants of miR-219-1 were not related to NSCLC in the Turkish population. The reason for this can be differences in ethnicity, regions, and background of population and these differences could lead to various outcomes. Keywords: NSCLC; miR-219-1 gene; single-nucleotide polymorphisms

Halkımızın teratoloji hakkındaki farkındalığı

Amaç: Bu çalışma ile çevresel etkenlerden kaynaklanan doğumsal bozuklukları ve anormal prenatal gelişmeyi inceleyen ve doğumsal defektleri önlemenin araştırmalarını yapan bilim dalı olan teratoloji hakkında halkımızın neler bildiğini ve konu ile ilgili farkındalıklarını araştırmayı amaçladık. Gereç ve Yöntem: Selçuk Üniversitesi Tıp Fakültesi Hastanesi’ne sağlık hizmeti almak üzere başvuran hasta ve hasta yakınlarından oluşan, yaşları 18 - 79 arasında değişen, eşit sayıda kadın ve erkekten oluşan toplam 400 katılımcıya anket uygulaması yapıldı. Elde edilen veriler ortalama ± standart sapma ve yüzde olarak özetlendi. Bulgular ve Sonuç: Yapılan anket çalışmasında homojen olmayan sonuçlar elde edildi. Bu verilere göre katılımcıların ölü ya da sakat doğumların sebepleri, röntgen, tomografi vb. kullanımının anne karnındaki bebeğe etkileri, ilaçların yan etkilerine dikkat etme, akrabalık derecesinin hastalıklı doğumlara etkisi konularında belirli düzeyde fikir sahibi oldukları saptandı. Anne karnındaki bebeğin hastalıklara karşı en duyarlı olduğu gebelik dönemi hakkında ise yeterli bilgi düzeyine sahip olmadıkları belirlend

Investigation of effects of some physical and chemical parameters on the heat shock response using recombinant Chlamydomonas reinhardtii strains

Bu çalışmada ısı şok yanıtında etkili olduğu düşünülen çeşitli kimyasal ve fiziksel parametreler moleküler araçlardan faydalanılarak C.reinhardtii üzerinde araştırılmıştır. Sürekli ekspresyona sahip 160 adet, ısıyla indüklenebilir ekspresyona sahip 96 tadet olmak üzere toplamda 256 adet transformant elde edilmiş ve bu transformantlar ARS aktivitesi bakımında kalitatif ve kantitatif olarak taranmıştır. Çalışmalara seçilen βTub II-32 ve HSP70A III-1 transformantları ile devam edilmiştir. Fiziksel parametre olarak sıcaklık, ışık şiddeti ve UV-B radyasyonun, kimyasal parametre olarak ise tuzluluğun, kalsiyumun ve kalsiyum şelatlayıcıları ve bloklayıcıların etkisi araştırılmıştır. Sıcaklığın etkisini incelemede hücreler 23 °C'den 35 °C, 37 °C, 40 °C ve 42 °C'ye kaydırılmış ve hücrelerin 35°C' ye dahi çıkarıldığında ısı şok yanıtının indüklendiği görülmüştür. Kalsiyumun 100 mM'ın üstünde ARS aktivitesini arttırdığı belirlenmiştir. Kalsiyum şelatlayıcıları olan EGTA ve BAPTA varlığında ısı stres yanıtının tetiklendiği gözlenmiştir. 100 µM lantan ve 250 µM gadolinyumun yükselen ARS aktivitesini düşürmede etkili olduğu saptanmıştır. Ancak verapamilin etki göstermediği belirlenmiştir. 2 000 lux ve üzeri ışık şiddetinin ise ARS aktivitesini arttığı saptanmıştır. UV-B uygulamasının ve tuzluluğun ARS aktivitesine herhangi bir etkisi bulunmamıştır. Tüm ARS enzim aktivite sonuçları RT-qPCR sonuçları ile karşılaştırmalı olarak değerlendirilmiş, paralel sonuçlar elde edildiği görülmüştür. Rekombinant C. reinhardtii ile elde edilen bu veriler bitki stresi ve ekolojik çalışmalar için önem taşıyabilir.In this study, various chemical and physical parameters which could be effective in heat shock response were investigated on C.reinhardtii by using molecular tools. In total 256 transformants were obtained, among them 160 transformants had continuous expression while 96 of them had heat-inducible expression and these transformants were screened for ARS activity qualitatively and quantitatively. Further studies were performed with selected transformants βTub II-32 and HSP70A III-1. As physical parameters the effect of temperature, light intensity and UV-B radiation; as chemical parameters salinity, calcium and calcium chelators and blockers were investigated. To investigate the effect of temperature, the cells were shifted from 23 °C to 35 °C, 37 °C, 40 °C and 42 °C. The heat shock response was induced even at 35 °C. It was determined that above 100mM of calcium concentration ARS activity was increased. In the presence of calcium chelators EGTA and BAPTA, heat stress response was triggered. 100µM lanthanum and 250 µM gadolinium were found effective in decreasing ARS activity. However, verapamil was not effective. It was found that light intensity of 2 000 lux and above increased ARS activity. UV-B and salinity had no effect on ARS activity. All ARS enzyme activity results were compared with the results of RT-qPCR and partially parallel results were obtained. These data obtained with recombinant C. reinhardtii might be important for plant stress and ecological studies

Perinatal outcomes and risk factors in adolescent and advanced age pregnancies: Comparison with normal reproductive age women

The objective of the study was to analyse and compare demographic characteristics and clinical outcomes of pregnancies in adolescent, advanced age and normal reproductive age women. All completed pregnancies in a 6-month period, registered by the family practitioners in Denizli province, were included into the study. A face-to-face questionnaire was used to gather information. Participants were asked for demographic information, pregnancy outcome and obstetric history, obstetric and neonatal problems. Overall 5,882 pregnancies in different age groups: 296 (5%) adolescent ( 35-years-old ) (group III), were included into the study. Adolescent women had a lower educational status (p < 0.01), and family played a major role in decision of marriage (p < 0.01). Birth weight of the baby was lower in adolescents (p < 0.01). While adolescents tended to deliver vaginally (OR = 1.9, p < 0.01), elderly women were more prone to operative delivery (OR = 1.2, p < 0.05). Risk of caesarean section rate was higher in elderly nulliparous women (OR = 2.2, p 0.01). The number of spontaneous and induced abortions were increased with age (p < 0.01). Antenatal problems were seen least frequently in normal reproductive age women. Both antenatal (OR = 1.7, p < 0.01) and neonatal problems (OR = 1.5, p < 0.05), were significantly higher in advanced age pregnancy. It was concluded that with sufficient antenatal care, adolescent pregnancy is not associated with an increase in adverse pregnancy outcome, except low birth weights. Advanced maternal age is more likely to be associated with increased obstetric, maternal and neonatal complications. © 2013 Informa UK, Ltd

Screening for latent tuberculosis in children with immune-mediated inflammatory diseases treated with anti-tumor necrosis factor therapy: Comparison of tuberculin skin and T-SPOT tuberculosis tests

Objectives: This study aims to analyze the coherence between T-SPOT tuberculosis test (T-SPOT.TB) and tuberculin skin test (TST) with different cut-off values in screening latent tuberculosis infection (LTBI) both prior to and at the sixth month of anti-tumor necrosis factor (anti-TNF) treatment.Patients and methods: This prospective multicentric study included 57 children (34 girls, 23 boys, mean age 12.4 +/- 3.9 years; range, 6 to 18 years) diagnosed with immune-mediated inflammatory diseases (IMIDs) evaluated with TST and T-SPOT.TB for screening LTBI both prior to and at the sixth month of treatment with anti-TNF agents. Coherence between two tests was analyzed for TST cut-off values suggested by the local guidelines and also for different possible cut-off values of TST.Results: Tuberculin skin test was positive (>= 5 mm) in 28.1% (n=16) of patients in the screening prior to treatment and in 33.3% (n=19) at the sixth month of treatment. T-SPOT.TB test was positive in 8.8% (n=5) of patients both prior to and at the sixth month of treatment. Coherence between two tests was poor or fair when compared with all possible TST cut-off values both prior to and at the sixth month of anti-TNF therapy.Conclusion: Our results show poor coherence between T-SPOT.TB and TST for all possible cut- off values of TST. Thus, using both tests would be beneficial in screening LTBI until further studies bring new evidence on the subject