Ümran Soyoğul Gürer

Potential Adjuvant Effects of Nigella sativa Seeds to Improve Specific Immunotherapy in Allergic Rhinitis Patients

Objective: To investigate the effects of Nigella sativa seed supplementation on symptom levels, polymorphonuclear leukocyte (PMN) functions, lymphocyte subsets and hematological parameters of allergic rhinitis. Subjects and Methods: Twenty-four patients randomly selected from an experimental group of 31 (mean age 34 years) sensitive to house dust mites with allergic rhinitis and a control group of 8 healthy volunteers (mean age 23 years) were treated with allergen-specific immunotherapy in conventional doses for 30 days. After a month of immunotherapy, 12 of the 24 patients and the 8 healthy volunteers were given N. sativa seed supplementation (2 g/day orally) for 30 days. The remaining 12 patients continued only on immunotherapy during the same period. The other 7 patients were given 0.1 ml saline solution subcutaneously once a week as a placebo. The symptom scores, PMN functions, lymphocyte subsets and other hematological parameters were evaluated before and after all treatment periods. Results: There was a statistically significant increase in the phagocytic and intracellular killing activities of PMNs of patients receiving specific immunotherapy, especially after the addition of N. sativa seed. The CD8 counts of patients receiving specific immunotherapy plus N. sativa seed supplementation significantly increased compared to patients receiving only specific immunotherapy. PMN functions of healthy volunteers significantly increased after N. sativa seed supplementation compared to baseline. Conclusion:N. sativa seed supplementation during specific immunotherapy of allergic rhinitis may be considered a potential adjuvant therapy.

Comparison of Polymorphonuclear Leukocyte Functions in Elderly Patients and Healthy Young Volunteers

Objective: To compare the polymorphonuclear leukocyte (PMN) functions (phagocytosis and intracellular killing activity) of elderly patients with healthy young volunteers. Subjects and Methods: Fifty-nine elderly patients who had various diseases (cancer, hypertension and diabetes mellitus, DM) and 10 healthy young volunteers were included in this study. Ficoll-Hypaque gradient centrifugation was used to isolate PMNs from venous blood containing EDTA (0.1 g/ml). Phagocytosis and intracellular killing activity of neutrophils were assayed using a modification of Alexander’s method, in which serum opsonins, number of neutrophils and number of microorganisms are standardized in order to detect both increases and decreases in phagocytosis and intracellular killing as well as combined abnormalities of these two functions. The least significant difference test was used to compare the results in the two groups. Results: Phagocytic activity of PMNs from patients with cancer was significantly higher than that of healthy young volunteers (p < 0.05) and elderly patients with hypertension and DM (p < 0.05). There was no statistical difference between the phagocytic activity of PMNs from elderly patients with hypertension and DM and healthy young volunteers (p > 0.05). The intracellular killing activity of PMNs from elderly patients with hypertension, DM and cancer was significantly lower than that of healthy young volunteers (p = 0.001, p < 0.0001, p = 0.003, respectively). Conclusions: The intracellular killing activity of PMNs from elderly patients was significantly decreased when compared with that of healthy young volunteers. Ageing, chronic diseases and drugs used in the treatment of these elderly patients may be the cause for decreased intracellular killing activity.

In vitro Effect of Amikacin, Imipenem, Cefodizime, IFNα-2a Alone and Combinations of Antibiotics with IFNα-2a on Polymorphonuclear Leukocyte Function in Chronic Hepatitis Patients

The in vitro effect of amikacin (8 µg/ml), imipenem (30 µg/ml), cefodizime (10 µg/ml), interferon α-2a (IFNα-2a) (10 IU/ml) and antibiotic combinations with IFNα-2a on polymorphonuclear leukocyte (PMN) functions (phagocytosis and intracellular killing of Candida albicans blastospores) was investigated in chronic hepatitis B patients. Phagocytosis and candidacidal activity was not affected after pretreatment of PMNs with amikacin and imipenem (p > 0.05). Phagocytic activity was enhanced after pretreatment of PMNs with cefodizime and IFNα-2a compared with that of control PMNs (p < 0.05), but candidacidal activity was not affected by the same drugs (p > 0.05). Phagocytic and intracellular activity of PMNs were not affected by combinations of IFNα-2a and antibiotics (p > 0.05).

Effect of montelukast on polymorphonuclear leukocyte functions in asthmatic patients.

Leukotriene receptor antagonists are being used widely in the treatment of bronchial asthma. They have been shown to possess anti-inflammatory properties, but there is no sufficient data about their effects on polymorphonuclear leukocyte functions. The aim of this study was to investigate the effects of montelukast, a specific cysteinyl leukotriene-1 receptor antagonist, on human polymorphonuclear leukocyte (PMN) functions (phagocytic and intracellular killing activity) in asthmatic patients. Fifteen mild to moderate asthmatic patients were included in the study. They were treated with montelukast (10 mg/day per os) in addition to their previous medications for 2 weeks. Whole blood samples of patients were taken before and after this treatment period. Phagocytic activities and intracellular killing activities of polymorphonuclear leukocytes isolated from whole blood samples were tested by using appropriate technics. Phagocytic and intracellular killing activities of PMNs were significantly increased (p<0.001, p<0.05) by montelukast compared to those before treatment. These results show that montelukast has an enhancing effect on PMN functions in asthmatic patients.

Effect of antipyretics on polymorphonuclear leukocyte functions in children

The aim of this study was to investigate whether fever and antipyretic drugs had an adverse effect on human polymorphonuclear leukocyte (PMN) functions (phagocytic and intracellular killing activity). Twenty febrile children with an axillary temperature of 39-40 degrees C and 20 healthy children without fever were included. Polymorphonuclear leukocytes were isolated. The effects of in vitro addition of antipyretic drugs (acetaminophen, metamizole sodium, nimesulid and ibuprofen) on PMN functions were tested. Phagocytic activity was assayed by the ingestion of yeast cells by PMNs and intracellular killing activity by the ingestion of yeast cells (stained blue) killed by PMNs. PMNs derived from febrile children exhibited better phagocytic activity when ibuprofen was added. In contrast, phagocytic activity was enhanced when acetaminophen, metamizole sodium or nimesulid was added in children without fever. Intracellular killing activity was enhanced when ibuprofen or metamizole sodium was added in children without fever. We conclude the antipyretic drugs at safely achievable concentrations do not suppress PMN function in vitro.

Bazı antibiyotiklerin Tip 2 diabetes mellitus’lu hastaların polimorfonüklear lökosit (PNL) fonksiyonları ve PNL’lerin miyeloperoksidaz aktivitesi, glutatyon ile malondialdehit düzeyleri üzerine in vitro etkisi

Amaç: Çalışmamızda diabetes mellitus (DM)’lu hastaların polimorf nüveli lökosit (PNL) fonksiyonları ile sağlıklı gönüllülerin miyeloperoksidaz (MPO) aktivitesi, glutatyon (GSH) ve malondialdehit (MDA) düzeylerini karşılaştırıp arasındaki ilişkiyi saptamayı amaçladık. Bunun yanında siprofloksasin, rifampisin ve doksisiklinin PNL fonksiyonları, MPO aktivitesi ve GSH ile MDA düzeyleri üzerine immunomodülatör etkilerini araştırdık. Yöntemler: Çalışmamızda Tip 2 diyabeti olan 13 hasta ve 13 sağlıklı gönüllü yer almaktadır. Antibiyotiklerin PNL fonksiyonları üzerine olan etkileri in vitro koşullarda araştırıldı. PNL’ler (1x107 hücre/ml) venöz kandan Ficoll-hypaque gradient santrifügasyon metodu kullanılarak izole edildi. PNL’lerin MPO aktivitesi modifiye O-dianisidin yöntemi, GSH miktarı Ellman yöntemi ve MDA düzeyleri ise Beuge yöntemi kullanılarak belirlendi. Bulgular: DM’lu hasta PNL’lerinin fagositik (p<0.05) ve hücre içi öldürme aktivitesi (p<0.01) sağlıklı gönüllülere göre anlamlı olarak azaldı. Siprofloksasin sağlıklı gönüllü ve tüm hasta PNL’lerinin fagositik ve hücre içi öldürme aktivitesini kontrol değerlere göre anlamlı olarak artırdı (p<0.01 ve p<0.01). Rifampisin, DM’lu hasta PNL’lerinin hücre içi öldürme aktivitesini anlamlı olarak artırdı (p<0.05). Rifampisin ve doksisiklin ise, sağlıklı gönüllü PNL’lerinin fagositik ve hücre içi öldürme aktivitesini kontrol değerlerine göre anlamlı olarak azalttı (p<0.05). Siprofloksasin (2.5 μg/ml) sağlıklı gönüllülerin (p<0.01) ve hastaların MPO aktivitesini ve MDA düzeyini ilaçsız kontrole göre anlamlı olarak artırdı (p<0.01). Ancak aynı ilaç DM’lu hasta PNL’lerinin GSH düzeyini anlamlı olarak azalttı (p<0.05). Sonuçlar: İmmün sistemi bozulmuş Tip 2 DM’lu hastalardaki ve sağlıklı gönüllerdeki MPO aktivitesi ile PNL fonksiyonları arasındaki ilişkinin önemi siprofloksasin ile hücresel immün sistemin desteklenmesi ve PNL’lerin MPO aktivitesi ile beraber PNL aktivitelerinin artışı ile gösterildi. Anahtar sözcükler: Diabetes mellitus, miyeloperoksidaz, fagositoz, hücre içi öldürme aktivitesi, antibiyotikler ABS TRACT The effect of some antibiotics on polymorphonuclear leukocyte (PMN) functions and PMN’s myeloperoxidase activity, glutathione and malondialdehyde levels of patients with type 2 diabetes mellitus in vitro

Bronşektazili hastalarda kolistinin tek başina ve tigesiklin, imipenem ve rifampisin ile kombinasyonlarinin polimorf nüveli lökosit fonksiyonlari, oksidatif stres, oksidan ve antioksidan enzimler üzerine etkilerinin in vitro araştirilmasi

OZET: Calismamizda, terapotik konsantrasyonlarda kolistinin (4 μg/ml) tek basina ve tigesiklin (0.87 μg/ml), imipenem (30 μg/ml) ve (rifampisin (7 μg/ml) ile kombinasyonlarinin bronsektazili hastalarin polimorf nuveli lokosit (PNL) fonksiyonlari (fagositik aktivite ve hucre ici oldurme aktivitesi) ve malondialdehit (MDA) duzeyi, superoksid dismutaz (SOD), glutatyon peroksidaz (GSH-Px) ve miyeloperoksidaz (MPO) aktiviteleri uzerine etkileri in vitro arastirilmistir. PNL’ler Ficoll-hypaque gradient santrifuj yontemi ile izole edilmis ve fagositik aktivite ve hucre ici oldurme aktivitesi tayini Alexander’in yontemi modifiye edilerek yapilmistir. Protein tayini Bradfort’un metodu kullanilarak, lipit peroksidasyonun gostergesi olarak malondialdehit (MDA) miktari Beuge’nin metodu, superoksit dismutaz (SOD) aktivitesi Sun ve arkadaslarinin metodu, glutatyon peroksidaz (GSH-Px) aktivitesi Paglia ve Valentine’nin metodu, miyeloperoksidaz (MPO) aktivitesi Hillegas ve arkadaslarinin metodu kullanilarak olculmustur. Tum sonuclar istatiksel olarak degerlendirilmistir. Bronsektazili hastalarda kolistin tek basina ve tigesiklin, imipenem ve rifampisin ile kombinasyonlari, fagositik aktivite (p<0.0001) ve hucre ici oldurme aktivitesini (p=0.0113, p=0.0008, p=0.0014, p=0.0036) kontrole gore anlamli olarak artirirken; MDA duzeyi, SOD, GSH-Px ve MPO aktivitelerini kontrole gore anlamli olarak azaltmistir (p<0.001). ANAHTAR KELIMELER: Antioksidanlar, bronsektazi, fagositoz, oksidatif stres

Flavonoids and Biological Activities of Centaurea stenolepis

Centaurea stenolepis Kerner (Asteraceae) is one of the 205 taxa of the genus Centaurea growing wild in Turkey [1]. To the best our knowledge, there is only one previous report on the essential oil of C. stenolepis [2]. The aim of present study was to isolate and characterize the flavonoids in the bioactive extract of C. stenolepis. Plant sample was collected in the flowering period from Catalca, Istanbul in 2009 and identified by Dr. Gizem Bulut (No. 11651). The dried and powdered stems and leaves of C. stenolepis (1 kg) were extracted with petroleum ether and ethanol with a Soxhlet extractor, respectively. The concentrated ethanol extract was diluted with water and extracted with toluene, chloroform, and ethyl acetate, respectively, for fractionation. The extracts were filtered and concentrated to dryness using a rotary evaporator to obtain toluene, chloroform, ethyl acetate, and aqueous ethanol extracts. From various fractions of ethyl acetate and petroleum ether extracts of C. stenolepis, by silica gel column chromatography, polyamide column chromatography, Sephadex LH-20 column chromatography, silica gel preparative circular chromatography (Chromatotron), silica gel preparative TLC, and paper chromatography (determination of sugars), we have isolated four flavonols, a flavonol aglycone in the petroleum ether extract, jaceidin (24 mg, 1), and three flavonol glucosides in the ethyl acetate extract, jacein (62 mg, 2), patuletin 3-O-glucoside (6.2 mg, 3), and quercetin 3-O-glucoside (9.4 mg, 4) 3, 4 . UV, NMR spectra (1H, 13C, HSQC, HMBC), and TLC-PC comparisons with authentic samples were used for the structure elucidation of the purified compounds. Jaceidin (1). C18H16O8 3, 5 . 1H NMR (500 MHz, CDCl3, , ppm, J/Hz): 3.88 (3H, s, 3-OCH3), 4.01 (3H, s, 6-OCH3), 4.07 (3H, s, 3 -OCH3), 7.70 (2H, H-2 , 6 ), 7.07 (1H, d, J = 8.2, H-5 ), 6.58 (1H, s, H-8). Jacein (2). C24H26O14 3, 6]. UV (MeOH, max, nm): 257, 272sh, 352; +NaOMe: 268, 408; +AlCl3: 270, 280sh, 295sh, 380; +AlCl3/HCl: 268, 280sh, 296sh, 369, 400sh; +NaOAc: 260, 359, 415; +NaOAc/H3BO3: 258, 270sh, 355. 1H NMR (500 MHz, DMSO-d6, , ppm, J/Hz): 3.88 (3H, s, 3 -OCH3), 3.84 (3H, s, 6-OCH3), 3.79 (3H, s, 3-OCH3), 7.67 (1H, d, J = 1.8, H-2 ), 7.62 (1H, dd, J = 1.8; 8.5, H-6 ), 6.99 (1H, d, J = 8.5, H-5 ), 7.03 (1H, s, H-8), 5.13 (1H, d, J = 7.4, Glc H-1 ), 3.82–3.18 (sugar protons). Patuletin 3-O-Glucoside (3). C22H22O14. 3, 7 . 13C NMR (125 MHz, DMSO-d6, , ppm): 177.4 (C-4), 155.87 (C-9), 153.5 (C-2), 152.15 (C-5), 151.71 (C-7), 148.51 (C-4 ), 144.84 (C-3 ), 132.89 (C-3), 131.60 (C-6), 121.50 (C-1 ), 121.20 (C-6 ), 116.12 (C-2 ), 115.23 (C-5 ), 104.33 (C-10), 101.01 (C-1 ), 94.01 (C-8), 77.49 (C-5 ), 76.51 (C-3 ), 74.09 (C-2 ), 69.91 (C-4 ), 60.95 (C-6 ), 59.77 (6-OCH3). The free radical scavenging capacity of various extracts of Centaurea stenolepis Kerner (toluene, chloroform, ethyl acetate, aqueous ethanol) and standard were evaluated according to the previously reported procedure using the stable DPPH [8]. Also, the total phenolic contents of these extracts were measured using Folin–Ciocalteau reagent [9]. The ethyl acetate extract was the most potent in antioxidant activity (IC50 value of the ethyl acetate extract 1.29 mg/mL) and total phenolic content (107.30 mg GAE/g dry material). Agar well diffusion and microdilution broth methods were used in the antimicrobial

Investigation of chronotherapeutic effects of amphotericin B administered to mice infected with Candida albicans

In this study, mice were infected with Candida albicans at 07:00 h or 19:00 h. After 24 h, the subgroups of mice received either 0.2/ml saline (as control) or one of two doses (0.5 or 1.0 mg/kg) of amphotericin B (AmB) at 0 h or 12 h for three consecutive days. A second set of uninfected mice received a single dose of either saline or AmB (5 mg/kg) at 0 h or 12 h for 4 days to study only about nephrotoxicity. For uninfected controls and AmB-treated (5 mg/kg) mice, serum blood urea nitrogen (BUN), creatinine and total protein tended to be higher at 0 h vs. 12 h, as was the histopathology score in treated mice (3.60 vs. 1.20). Serum levels changed in treated mice when compared to the control mice. The BUN levels increased whereas serum creatinine levels decreased at 12 h compared to 0 h. C. albicans colony forming units per milliliter (CFU/ml) was high in the kidneys of infected mice. Compared with the control, after treatment for 3 days with 0.5 mg AmB lowered CFU by 48% at 0 h and by 75% at 12 h. However, for the higher dose 1.0 mg AmB, CFU was lowered more or less equally at both test times: 51% at 0 h and 46% at 12 h.