Vibrio vulnificus infection in horse mackerel (Trachurus mediterraneus) and its characteristic symptoms: An experimental study

In the present study, the mortality and characteristic symptoms (bilateral exophthalmia and skin lesions) associated with Vibrio vulnificus infection in horse mackerel (Trachurus mediterraneus) are presented from an experimental infection trial

Outcomes of Therapeutic and Tectonic Penetrating Keratoplasty in Eyes with Perforated Infectious Corneal Ulcer

Objectives:To evaluate the outcomes of penetrating keratoplasty performed for therapeutic and tectonic purposes in eyes with perforated infectious corneal ulcer.Materials and Methods:This retrospective study included 43 eyes of 43 patients who developed perforated infectious corneal ulcer of various etiological causes between June 2008 and January 2018. The patients were evaluated based on age and sex, follow-up time, presence of corneal perforation, pre- and postoperative visual acuity, postoperative graft transparency, complications, and infection recurrence.Results:The mean age of the 43 patients was 52.9±13.8 years. The mean follow-up time was 2.7±1.3 years. Preoperatively, the visual acuity of the eyes was at the level of hand motions or counting fingers; postoperative best corrected visual acuity ranged from hand motions to 0.7. Postoperative complications included hyphema in 8 patients (18.6%), elevated intraocular pressure in 14 (32.5%), posterior synechiae in 18 (41.8%), and cataract in 22 patients (51%). Therapeutic and tectonic success was achieved in 42 patients (97.6%). Postoperative graft transparency was observed in 35 patients (83.3%) within the 1-year follow-up period and in 27 patients (71.0%) at 2 years. Among 27 patients with graft transparency, 23 had bacterial and 4 had viral etiologies (p=0.52); 16 patients had perforations smaller than 1 mm and 11 had perforations 1-3 mm in size (p=0.2).Conclusion:Therapeutic-tectonic keratoplasty for perforated infectious corneal ulcer successfully restored globe integrity in 97.6% of cases. The rate of graft transparency was 71.0% at 2 years, with no effect of etiological agent or perforation size

In Vitro Regulation of the Expression of the Sars-Cov-2 Receptor Angiotensin-Converting Enzyme (Ace2) in Lung Cancer Cells by Natural Products

DergiPark: 896013trkjnatThe COVID-19 pandemic continues infecting people causing deaths globally. Although various medicines have been tried to combat with COVID-19, there is no medicine or treatment that has been validated yet. People have been using natural products for centuries against bacterial and viral illnesses. This study aimed to test the effects of the biomolecule oleuropein, whey collected from industrial waste and soaproot extracts obtained from Gypsophila arrostii Guss. var. nebulosa Boiss. amp; Heldr. and Saponaria officinalis L. on the expression of the human ACE2 gene as SARS-CoV-2 receptor on the A549 adenocarcinoma cell-line by Real-Time Quantitative Polymerase Chain Reaction (qPCR). According to the cytotoxicity tests, G. arrostii var. nebulosa and S. officinalis extract treatments showed a dose dependent cytotoxic effect on the cells. The EC50 values of G. arrostii var. nebulosa and S. officinalis were found to be 54.3 ?g/ml and 17.3 ?g/ml, respectively. Oleuropein showed moderate cytotoxic effects with the EC50 value over 250 ?g/ml. Whey (fermented and non-fermented) did not show any cytotoxic effect at the applied doses. The qPCR results showed that the ACE2 mRNA level decreased by 89.8% and 35.2% due to the fermented and non-fermented whey extracts, respectively. Similarly, G. arrostii var. nebulosa and S. officinalis downregulated ACE2 by 79.8% and 90.1%, respectively. In contrast, oleropein upregulated ACE2 (102.8%). Our results showed that the natural supporting products produced from soaproot extracts and fermented whey can be used against COVID-19 by both cancer patients and people in potential risk groups. COVID-19 pandemisi tüm dünyada küresel çapta insanları enfekte etmeye ve ölümlere neden olmaya devam etmektedir. COVID-19 ile mücadelede birçok ilaç denenmiş olmasına karşın henüz herhangi bir ilaç veya tedavi yöntemi onaylanmamıştır. İnsanlar yüzyıllardan bu yana hastalıklara karşı doğal ürünleri kullanmışlardır. Bu çalışmadaki amacımız bir biyomolekül olan oleuropein, endüstriyel atık olarak bertaraf edilen peynir altı suyu ve Gypsophila arrostii Guss. var. nebulosa Boiss. amp; Heldr. ve Saponaria officinalis L. bitkilerinden elde edilen ekstraktların A549 kanserli hücre hatlarında ACE2 reseptörünü kodlayan ACE2 geninin anlatım seviyesi üzerine etkilerini Gerçek Zamanlı Kantitatif Polimeraz Zincir Reaksiyonu (qPCR) ile belirlemektedir. Yaptığımız sitotoksisite testlerine göre G. arrostii var. nebulosa ve S. officinalis ekstraktları sırası ile 54,3 ?g/ml ve 17,3 ?g/ml EC50 değerleri ile doza bağımlı sitotoksik etki gösterirken, oleuropein 250 ?g/ml’nin üzerinde bir değerle orta dereceli sitotoksik etki göstermiştir. Öte yandan peynir altı suyu (ferment eve fermente edilmeyen), çalışmada kullanılan dozlarda sitotoksik etki göstermemiştir. qPCR sonuçlarına göre fermente edilmiş ve edilmemiş peynir altı suyunun ACE2 genine ait mRNA seviyesini sırası ile %89,8 ve %35,2 oranlarında düşürdüğü belirlenmiştir. Benzer şekilde G. arrostii var. nebulosa ve S. officinalis ekstraktlarının ACE2 geni mRNA seviyesini sırası ile %79,8 ve %90,1 oranında düşürdüğü belirlenmiştir. Bu sonuçların aksine oleuropein biyomolekülünün ACE2 mRNA seviyesini %102,8 oranında arttırdığı belirlenmiştir. Çalışma sonuçlarına göre kullanılan bitki ekstraktlarının ve fermente edilmiş peynir altı suyunun COVID-19 ile mücadelede kanser hastalarında ve risk gruplarında kullanılabilecek doğal destek ürünlerinin üretilmesinde kullanılabileceğini göstermektedir

Diyabetik tavşanlarda sitokromp 450'ye bağımlı ilaç metabolizması

Diabetus mellitus is a metabolic disorder of carbohydrate, fat and protein metabolism as a result of insulin deficiency or diminished tissue response to insulin. In this study, diabetes was induced in rabbits experimentally by intravenous injection of 100mg/kg alloxan. Injection of a single dose of alloxan to rabbits induced diabetus mellitus as it was understood from 4-fold increase in the blood glucose level and also 2-fold increase in blood urea concentrations. The influence of diabetus mellitus on cytochrome P450 levels, microsomal cytochrome P450 dependent drug metabolizing enzymes and on the biomarkers used to measure chemical-induced toxicity including AST, ALT and LDH were examined in rabbit liver, kidney and lung.Results obtained in this study showed that diabetus mellitus elevated microsomal cytochrome P450 contents of liver and kidney by 1.3- and 2-fold, respectively. Induction of the cytochrome P450 isozymes was observed with the high intensity bands having Mr between 51 000 Da and 53 000 Da and 45 000 Da to 48 000 Da in the SDS-PAGE profiles of liver microsomes obtained from the diabetic rabbits. However, the changes in kidney and lung microsomal protein patterns of diabetic rabbits compared to control animals were not so pronounced as observed in rabbit liver. Induction of diabetus mellitus caused 1.67-, 2.36- and 1.32-fold increases in aniline 4-hydroxylation rates of liver, kidney and lung microsomes, respectively. Induction of experimental diabetes enhanced hydroxylation rates of /Miitrophenol by liver, kidney and lung microsomes about 1.8-, 2.3- and 1.32-fold, respectively. NDMA N- demethylase activity was enhanced by diabetes in all three of these organs. (1.98-fold in liver, 1.85-fold in kidney, 1.35-fold in lung). Induction of diabetes in rabbits caused an increase of the P4502E1 level of microsomes and this was reflected in increased rate of metabolism of aniline, p-nitrophenol and NDMA. In this study, blood serum triglyceride, creatinine, urea and cholesterol concentrations were determined in control and diabetic rabbits. In addition, the effects of diabetes on transaminases (ALT and AST) and lactate dehydrogenase enzyme activities of rabbit blood serum and soluble fractions of rabbit liver, kidney and lung were investigated. Histological studies were carried out in control and diabetic rabbit liver and kidney tissue samples to investigate any tissue degeneration (necrosis). This is the first time induction of cytochrome P450 dependent monooxygenases has been reported for liver, kidney and lung microsomes of diabetic rabbits.Diabetus mellitus, insulin hormonu eksikliği yada dokuların insuline cevabında azalma nedeniyle ortaya çıkan, karbonhidrat, lipit ve protein metabolizmasındaki değişimlerle karakterize edilen kronik bir hastalıktır. Bu çalışmada, tavşanlarda, damar içine 100 mg/kg alloksan verilmek suretiyle deneysel olarak diyabet oluşturuldu. Tek doz alloksan enjeksiyonunun diyabete neden olduğu, kanda glikoz seviyesinin 4-kat artışı ve aynı zamanda kandaki üre konsantrasyonunun 2-kat artışı ile gösterilmiştir. Diabetus mellitusun, sitokrom P450 seviyeleri, mikrozomal P450'ye bağımlı ilaç metabolize eden enzimler ve AST, ALT ve LDH gibi kimyasal toksisite ölçümünde kullanılan biyobelirteçler üzerinde gösterdiği etkileri tavşan karaciğer, böbrek ve akciğerinde incelendi.Bu çalışmada elde edilen sonuçlar, Diabetus mellitus'un; karaciğer ve böbrek mikrosomal sitokrom P450 miktarlarını sırasıyla 1.3- ve 2-kat arttırdığını gösterdi. Sitokrom P450 izozimlerinin indüksiyonu, diyabet oluşturulmuş tavşanlardan elde edilen karaciğer mikrozomlannın SDS-PAGE profillerinde yaklaşık olarak 51 000 Da ile 53 000 Da ve 45 000 Da ile 48 000 Da molekül ağırlıkları arasına denk gelen bölgelerdeki koyu bandlar ile gözlendi. Ama, diyabetik tavşanların böbrek ve akciğer mikrozomal protein profilleri kontrol tavşanları ile karşılaştırıldığında, karaciğer kadar farklı olmadığı görüldü. Diabetus mellitusun indüksiyonu, karaciğer, böbrek ve akciğer mikrozomlanndaki anilin 4 -hidroksilaz enzim aktivitesinin sırasıyla 1,67-, 2.36 ve 1.32-kat artmasına neden oldu. Diyabetin oluşumu /7-nitrofenolün karaciğer, böbrek ve akciğer mikrozomlan tarafından hidroksilasyon hızım, farkedilir bir şekilde; sırasıyla 1.8-, 2.3- ve 1.32-kat arttırdı. Diyabet bu üç organda da NDMA N-demetilaz aktivitesini arttırdı. (Karaciğerde 1.98-kat, böbrekte 1.85-kat, akciğerde 1.35-kat). Tavşanlarda diyabetin oluşturulması, mikrozomlann P4502E1 seviyesinde bir artışa neden oldu ve bu artış anilin, /?-nitrofenol ve NDMA metabolizmasındaki artış şeklinde de gözlendi. Bu çalışmada, kan serum trigliserid, kreatin, üre ve kollesterol konsantrasyonları diyabetik ve kontrol tavşanlarda belirlendi. Buna ek olarak, diyabetin serum ve karaciğer, böbrek ve akciğer transaminazlar (ALT ve AST) ve laktat dehidrojenaz enzim aktiviteleri üzerine etkileri de incelendi. Histolojik çalışmalar, kontrol ve diyabetik tavşan karaciğer ve böbrek doku örneklerinde herhangi bir dejenerasyon (nekroz) olup olmadığım incelemek için yapıldı. Diyabetik tavşanların karaciğer, böbrek ve akciğer mikrozomlanndaki sitokrom P450 bağımlı monooksijenazlann indüksiyonu ilk kez bu çalışmada gösterildi.M.S. - Master of Scienc

Diyabetli tavşanlarda Benzen'in karaciğer, börek ve akciğer cyp1a, cyp2b4, cyp2e1 ve cyp3a6 mrna, protein seviyesi ve ilaç metabolize eden enzim aktiviteleri ve toksisite üzerine etkileri.

The effects of diabetes on cytochrome P450 dependent drug metabolizing enzymes have not to be clarified yet. The most widely used animals in these studies have been rats, and information regarding the effects of diabetes on cytochrome P450 dependent procarcinogen/carcinogen metabolism in rabbits is limited. In the present study, we investigated, for the first time, the influence of benzene on liver, kidney and lung microsomal cytochrome P450 dependent drug metabolizing enzyme activities, protein and mRNA levels in diabetic and non-diabetic rabbits. Male New Zealand rabbits were made diabetic by a single dose of alloxan treatment in this study. AST, ALT and LDH enzyme activities in the blood serum and lipid peroxidation in liver microsomes were found to increase in diabetic, benzene treated and benzene treated diabetic rabbits. Besides these, CYP2E1 dependent NDMA N-demethylase and p-nitrophenol hydroxylase activities and CYP2E1 protein level were found to increase in liver and kidney of diabetic and benzene-treated rabbits. The combined effects of benzene and diabetes on these activities and protein level were found to be additive. Although diabetes caused induction of pulmonary CYP2E1 protein level and associated enzyme activities, benzene treatment of rabbits resulted in no change in enzyme activities and protein level in lung. The level of mRNA was investigated by Real-Time PCR. Accordingly, hepatic CYP2E1 mRNA level was increased 6.71-, 10.53- and 12.93-fold in diabetic, benzene treated and benzene treated diabetic rabbits with respect to the control animals. Similarly, renal CYP2E1 mRNA level was found in increase in these rabbits. In addition to CYP2E1, CYP3A6 associated enzyme activity, erythromycin N-demethylase, CYP3A6 protein and mRNA level were found to increase in diabetic rabbit liver and lung. Unlike diabetes, benzene treatment caused suppression of CYP3A6 protein and inhibition of associated enzyme activity in liver. There was no significant change in the erythromycin N-demethylase activity and CYP3A6 level of liver and lung as a result of benzene treatment of diabetic rabbits. Moreover, diabetes induced CYP1A2 protein and mRNA level and CYP1A associated enzyme activities in the rabbit liver. On the other hand, benzene caused statistically insignificant decreases in CYP1A dependent enzyme activities and CYP1A2 protein level in liver. CYP1A associated enzyme activities, CYP1A2 protein and mRNA levels were not changed in the liver of benzene treated diabetics. The results of the present work indicate that both diabetes and benzene stimulate metabolic activation toxic chemicals metabolized by CYP2E1 such as NDMA and benzene by inducing CYP2E1 which results in the formation of increased amounts of reactive metabolites. Application of benzene to diabetic rabbits further elevates expression and activities of the CYP2E1. As a result of additive induction of the CYP2E1 in benzene treated diabetics, further increase the risk of hepatotoxicity produced by toxins may be observed when compared to the separate treatments. This may in turn further potentiate the risk of organ toxicity and mutagenesis in liver and kidney of these subjects. As in the case of CYP2E1, the risk of carcinogenesis due to induction of CYP1A may be increased in diabetic subjects. Moreover, in diabetic and benzene exposed subjects, alteration of drug clearance and clinical drug toxicity may be observed due to induction or suppression of CYP3A.Ph.D. - Doctoral Progra

Effects of Acrylamide on Cytohrome P450 Dependent Drug metabolizing Enzymes in Human Liver Cell Line (HepG2).

Başlangıç seviyesi projesiEndüstrileşmenin gelişmesiyle birlikte çevreye her yıl binlerce çeşit kimyasal madde atılmaktadır. Buna ilave olarak, teknolojinin ilerlemesi ve tıp alanındaki gelişmelerle birlikte her gün yeni bir ilaç çeşitli hastalıkların tedavisi amacıyla üretilmektedir. Bunun sonucu olarak, dünyamızdaki canlılar ve özellikle insanlar, birçoğu toksik olan bu bileşiklere ve kimyasallara maruz kalmaktadırlar. Bu kimyasallardan en önemlilerinden biri akrilamidtir. Akrilamid birçok endüstride sıklıkla kullanılan, nörotoksik, reprodüktif toksik ve karsinojenik bir kimyasaldır. Akrilamid aynı zamanda yüksek ısıda pişirilmiş karbohidratlı gıdalarda oluştuğu için insanların kolaylıkla diyetlerinde maruz kalabilecekleri bir kimyasaldır. Çalışmalar akrilamidin mutajenik, genotoksik, reaktif bir ürün olan glisidamide, sitokrom P450’ye bağımlı monooksijenaz enzimlerinin izozimlerinden P4502E1 (CYP2E1) tarafından dönüştürüldüğünü göstermiş ve bu metabolik yol akrilamidin neden olduğu karsinojenisite ile ilişkilendirilmiştir. Ancak, akrilamidin CYP2E1 enzimi ve diğer P450’ye bağlı sitokrom önkarsinojen/karsinojen metabolizmasına olan etkilerini araştıran çalışmalar sınırlı sayıdadır. İnsan karaciğer hücre kültüründe ise böyle bir çalışmaya literatürde rastlanmamıştır. Çeşitli alanlarda yaygın kullanımı, bazı ürünlerin yapısında bulunması ve vücuttaki metabolizması sonucu oluşan metabolitinin karsinojenik olması, akrilamidi insan sağlığı açısından önemli yapmaktadır. Bu nedenle, bu calışmanın amacı, akrilamidin P450’ye bağımlı ilaçları metabolize eden bazı enzimler üzerine olan etkisinin insan karaciğer kanser hücre dizisinde (HepG2) aydınlatılmasıdır. Bu amaçla; değişik konsantrasyonlarda akrilamid kullanılarak akrilamidin sitotoksik etkisi HepG2 hücre hattında saptandı. Elde edilen sitotoksisite sonuçlara göre, 1.25 mM ve 2.5 mM akrilamid konsantrasyonu seçilerek, sitokrom P450 izozimlerinin nasıl etkilendiği belirlendi. İki farklı konsantrasyonda akrilamid uygulanması, CYP1A1 ve CYP1A2’ ye bağlı etoksiresorufin O-deetilaz ve metoksiresorufin O-demetilaz aktiviteleri ile bu izozimlerin protein ve mRNA seviyelerinde istatistiksel olarak anlamlı bir artışa sebep oldu. (p<0.05). Benzer şekilde, CYP2E1 bağlı anilin 4-hidroksilaz aktivitesi ile protein ve mRNA seviyeleri akrilamid uygulanması sonucunda arttığı bulundu. Bunların aksine, CYP3A4’e bağlı eritromisin N-demetilaz aktivitesi ve protein seviyesi her iki konsantrasyonda herhangi bir değişikliğe sebep olmazken, aynı izozimin mRNA seviyesi konsantrasyona bağımlı bir şekilde azalmıştır. Bu veriler ışığında, sitokrom P450 izozimlerinin akrilamid uygulaması ile hem aktivite, hem de protein ve mRNA seviyelerinin değişmesi sonucunda, bu kimyasala maruz kalan insanlarda kanser oluşum riski, klinik toksisite ve ilaç metabolizmasında bozuklukların görülme olasılığı artabilir.In each year, thousands of different chemicals are released into environment with industrial development. In addition to this, in each day, a new drug is produced for treatment of various diseases in parallel to the technological improvement and medicinal development. In consequence of this, organisms in earth, especially humans, are exposed to the these mostly toxic chemicals. One of the most important chemical among these chemicals is acrylamide. Acrylamide is one of the most widely used chemical in various industrial sites with its known neurotoxic, reproductive toxin and carcinogenic effects. Humans are exposed to acrylamide simply in their diet due to formation of acrylamide in carbohydrate rich foods cooked at higher temperatures. Studies have demonstrated that acrylamide is metabolized to mutagenic, genotoxic and carcinogenic metabolite, glycidamide by P4502E1 (CYP2E1), one of the isozyme of cytochrome P450 dependent monooxygenase enzymes, and the carcinogenicity associated with acrylamide is mostly attributed to this metabolism. However, studies regarding the effects of acrylamide on CYP2E1 enzyme and on other cytochrome P450 dependent procarcinogen/carcinogen metabolism are limited. There are no available studies related to the effects of acrylamide on this system in human liver cell culture in literature. Due to its wide usage, presence of in some products, and its carcinogenic metabolite formed as a result of metabolism in body, acrylamide is turned out to be very important for human health. In this regard, the aim of this study is to elucidate the effects of acrylamide on cytochrome P450 dependent drug metabolizing enzymes in human hepatoma cell line (HepG2). For this purpose, cytotoxic effect of acrylamide was determined in HepG2 cell line by using various concentrations of this chemical. The effects of acrylamide on cytochrome P450 isozymes were determined by choosing 1.25mM and 2.5mM acrylamide concentration according to the cytotoxicity results. Acrylamide treatment in two different concentrations was caused statistically significant increase in CYP1A1 and CYP1A2 associated ethoxyresorufine O- deethylase and methoxyresorufine O-demethylase activities and protein and mRNA levels of these isozymes (p<0.05). Similarly, it was found that CYP2E1 associated aniline 4- hydroxylase activity and protein and mRNA levels increased by acrylamide treatment. On the other hand, although CYP3A4 associated erytromycin N-demethylase activity and CYPA4 protein level was not changed significantly, the mRNA level of this isozyme was decreased in concentration dependent manner. In the light of these data, in acrylamide exposed people, the probability of cancer formation risk, clinical toxicity and disturbance in drug metabolism may be increased due to changes in P450 izozymes at both activity and protein and mRNA levels by acrylamide treatmen