False Positive Diagnosis of Lysosomal Storage Disease Based on Dried Blood Spot Sample; Leucocyte Number of a Challenging Factor

Aim:Recently dried blood spot (DBS) samples have been recommended as a screening test for Lysosomal Storage diseases. Although DBS samples have many advantages including non-invasiveness, cost and transportation, usage of these samples is limited by its high false positive rate. We aimed to investigate any possible effect of the leucocyte number on enzyme activity in dried blood samples in a retrospective study.Materials and Methods:Data was collected from subjects (n=263) for whom hematological parameters were available in the database of Ege University Hospital. The lysosomal enzyme activity results (alpha glycosidase, glycocerebrosidase, alpha galactosidase, sphingomyelinase and galactocerebrosidase) were re-evaluated with regard to the leucocyte number. Enzyme activities were measured using fluorometric and liquid chromatography-tandem mass spectrometry methods.Results:All enzyme activities closely correlated with the total number of leucocyte, since leucocytes are the main source of lysosomal enzymes. Glycocerebrosidase and galactocerebrosidase presented a positive correlation with the number of neutrophils and sphingomyelinase showed a positive correlation with the number of lymphocytes. When we recalculated the lysosomal enzyme activities with regard to the leucocyte number, the false positive rates for glycocerebrosidase, sphingomyelinase and alpha galactosidase decreased from 20%, 10.5% and 10.8% to 4.5%, 4.4% and 4.2%, respectively.Conclusions:Our data indicated that the enzyme activity in dried blood samples including low leucocyte number might be found lower than reference intervals resulting in false positive diagnosis. We concluded that the calculation of enzyme activity with regard to the number of leucocytes might produce more reliable results and might be helpful in decreasing the false positive rate

Immunohistochemical Examination on the Effects of Malathion and Onosma Nigricaule (Boraginaceae) on the Leptin in Renal Tissues of Mice

AimThe purpose of this study was to determine the effects of the plant extract, which is obtained from Onosma nigricaule due to the oxidation parameters caused in mice by Malathion that is used as an insecticide in agriculture, on leptin in renal tissues in an immunohistochemical way.Material-MethodsA total of 48 male mice were used in our study. 6 groups were determined as the control (group 1), maize oil which is the transport agent of malathion (group 2), normal saline which is the transport agent of plant extract (group 3), Onosma nigricaule (group 4), malathion (group 5) and Onosma nigricaule plant extract + malathion (group 6) group. The localization of leptin in the renal tissue was determined using the method of Streptavidin-Biotin- Peroxidase. Hematoxylin-eosin and triple staining methods were used for histological and pathological examinations.ResultsNo lesion was determined in renal tissues of the control group, normal saline group, maize oil group, Onosma nigricaule malathion and onosma nigricaule + malathion groups. Leptin immunoreactivity was determined with a weak intensity in renal tubulus proximalis and distalis epithelial cells of mice in the onosma nigricaule group, and onosma nigricaule + malathion group and with a high intensity in tubulus proximalis of other groups.ConclusionThis study conducted on mice revealed that malathion, which is commonly used as insecticide in Turkey, affects leptin activities in renal tissue of mice

The effects of kefi r, koumiss, yogurt and commercial probiotic formulations on PPAR a and PPAR- b / d expressions in mouse kidney

Commercial probiotic capsules that contain probiotic bacteria, kefir, koumiss and yogurt contain beneficial microorganisms that affect cholesterol levels and immune response, and are used for treatment of some diseases. We investigated using immunohistochemistry the effects of kefir, koumiss, yogurt and a commercial probiotic formulation on the expression levels of peroxisome proliferator-Activated receptor-? (PPAR?) and peroxisome proliferator-Activated receptor-?/? (PPAR-?/?), which are members of the nuclear steroid hormone receptor superfamily in mouse kidney. Mice were assigned to five groups: group 1, commercial probiotic capsules; group 2, kefir; group 3, koumiss; group 4, yogurt; group 5, control. After oral administration for 15 days, body weights were recorded and kidney tissue samples were obtained. Hematoxylin & eosin staining and the streptavidin-biotin peroxidase complex (ABC) method were applied to tissue sections to examine histology and to determine the localization of PPAR? and PPAR-?/? in the kidneys. We found that the weights of the mice in the kefir, koumiss, yogurt and commercial probiotic capsules groups were increased compared to controls. No differences in kidney histology were observed in any of the experimental groups. Kefir, koumiss, yogurt and the commercial probiotic preparation increased PPAR? and PPAR-?/? expressions. © 2014 The Biological Stain Commission.This Project was supported by the Commission for the Scientific Research Projects of Kafkas University, project number 2012-VF-5 Declaration of interest: The authors report no conflicts of interest. The authors alone are responsible for the content and writing of the paper

Immunohistochemical Examination on the Effects of Malathion and Onosma Nigricaule (Boraginaceae) on the Leptin in Renal Tissues of Mice

Amaç Bu çalışmada, tarımda bir insektisit olarak kullanılan malathion’un neden olduğu oksidasyon parametreleri üzerine Onosma nigricaule (Boraginaceae)’den elde edilen bitkisel özütün böbrek dokusunda leptin üzerine etkisinin immunohistokimyasal olarak araştırılması amaçlandı. Materyal-Metot Çalışmamızda, toplam 48 adet erkek fare kullanıldı. Gruplar; kontrol (grup 1), malathion’un taşıt maddesi olan mısır yağı (grup 2), bitki özütünün taşıt maddesi olan serum fizyolojik (grup 3), malathion (grup 4), Onosma nigricaule (grup 5), ve Onosma nigricaule + malathion grubu (grup 6) olmak üzere toplam 6 grup olarak belirlendi. Böbreklerden alınan kesitlere leptin immunoreaktivitesini belirlemek için Streptavidin-Biotin-Peroksidaz Kompleks tekniği uygulandı. Hazırlanan kesitlere histolojik incelemeler için hematoksilen-eosin ve Crossman’ın üçlü boyama yöntemi kullanıldı. Bulgular Kontrol grubu ile taşıt kontrol grupları olan serum fizyolojik grubu ve mısır yağı grupları ile Onosma nigricaule, malathion ve Onosma nigricaule + malathion gruplarının böbrek dokularında herhangi bir lezyon tespit edilmedi. Diğer yandan, kontrol, mısır yağı, serum ve malathion grubundaki farelerin tubulus proksimalis ve tubulus distalis epitel hücrelerinde güçlü leptin immunoreaktivite görülmesine karşın Onosma nigricaule ve Onosma nigricaule + malathion grubu farelerde ise zayıf leptin immunoreaktivitesi tespit edildi. Sonuç Fareler ile yaptığımız çalışmada, yurdumuzda insektisit olarak yaygın kullanılan malathion’un fare böbreğinde leptin aktivitelerini etkilediği görülmüştür.Aim The purpose of this study was to determine the effects of the plant extract, which is obtained from Onosma nigricaule due to the oxidation parameters caused in mice by Malathion that is used as an insecticide in agriculture, on leptin in renal tissues in an immunohistochemical way. Material-Methods A total of 48 male mice were used in our study. 6 groups were determined as the control (group 1), maize oil which is the transport agent of malathion (group 2), normal saline which is the transport agent of plant extract (group 3), Onosma nigricaule (group 4), malathion (group 5) and Onosma nigricaule plant extract + malathion (group 6) group. The localization of leptin in the renal tissue was determined using the method of Streptavidin-BiotinPeroxidase. Hematoxylin-eosin and triple staining methods were used for histological and pathological examinations. Results No lesion was determined in renal tissues of the control group, normal saline group, maize oil group, Onosma nigricaule malathion and onosma nigricaule + malathion groups. Leptin immunoreactivity was determined with a weak intensity in renal tubulus proximalis and distalis epithelial cells of mice in the onosma nigricaule group, and onosma nigricaule + malathion group and with a high intensity in tubulus proximalis of other groups. Conclusion This study conducted on mice revealed that malathion, which is commonly used as insecticide in Turkey, affects leptin activities in renal tissue of mice

Immunohistochemical Examination on the Effects of Malathion and Onosma Nigricaule (Boraginaceae) on the Leptin in Renal Tissues of Mice

[Abstract Not Available

Aşil tendon rüptürlerinde yeni bir cerrahi yöntem: tavşan aşil tendon rupturlarının immobilizasyon yapılmaksızın implant İle onarımı ve geleneksel dikiş yöntemleriyle karşılaştırılması

Purpose of this study was to repair achilles tendon rupture with protection of the integrity of the tendon without the need for the joint immobilization and prevent the complications reported with other techniques. For this purpose, mostly used techniques, Krackow, Modified Kessleer, Bunnel techniques were compared with the newly developed technique using the objective histologic and biomechanic criteria. Depending on the preliminary results of the pull-out tests, 6 tendons were included at each group, consisting of four groups totally. Operations were performed under the anesthesia of xylazine hydrochlorur, bupivacain HCI. At group 1, imlant prepared from the flexible polyethylene material was used. Krackow technique at group 2, Modified Kessler technique at group 3, Bunnell technique at group 4 were utilized. Rabbits were euthanised at 6th week postoperatively by use of overdose of sodium pentobarbital (50 mg/kg). While there was no difference between groups at histological analysis, at biomechanical analysis ultimate load to failure was found to be higher than all other techniques. Depending on these results, newly described technique can be applied clinically and may help to solve this orthopaedic problem.Bu çalışmayla eklemleri immobilize etme gereği olmadan ve tendon bütünlüğü mümkün olduğunca korunarak aşil tendon rüptürlerinin onarılması ve diğer tekniklerde bildirilen komplikasyonların önemli bir bölümünün önlenmesi amaçlandı. Bunun için günümüzde en sık kullanılan Krackow, Modifiye Kessler ve Bunnell sütür tekniklerinin uygulandığı üç ayrı kontrol grubu ve geliştirilen tekniğe ait sonuçlar biyomekanik objektif kriterlere göre karşılaştırılması yapıldı. Demonstrasyon olgularından elde edilen çekme deneylerinin sonuçları dikkate alınarak her grupta 6 denek olacak şekilde 4 ayrı grup oluşturuldu. Operasyonlar xylazine hydroklorür (Rompun 2% -Bayer) sedasyonunu izleyerek bupivacain HCI ( Marcaine 0,05- Eczacıbaşı) ile elde edilen spinal anestezi altında gerçekleştirildi. I. Grupta üzerinde delikler açılarak önceden hazırlanan fleksibl polyetilen materyalden hazırlanmış implant kullanıldı. II. grupta Krackow, III. grupta Modifiye Kessler, IV. grupta ise Bunnell dikiş teknikleri uygulanarak rüptür onarıldı. Denekler postoperatif 6.haftada overdose sodium penthobarbital (50 mg/kg) kullanılarak ötenazi edildi. Histopatolojik açıdan gruplar arasında önemli bir fark saptanamazken, biyomekanik test sonuçları dikkate alındığında geliştirilen tekniğe göre hem tendonun maksimum dayanacağı yük hem de uzama miktarı literatürdeki bütün tekniklere göre oldukça yüksek bulundu. Bu sonuç geliştirilen tekniğin maksimum dayanım sınırından sonra bile belirli bir süre daha yük taşımaya devam ettiğini göstermektedir. Buradan elde edilen sonuçlardan aynı tekniğin insan modellerinde de uygulanabileceği kanısına varılırken, tekniğin insan modellerinde başarılı olması durumunda bu alanda çok önemli bir ortopedik problemin sorunsuz bir şekilde çözüme kavuşturulabileceği söylenebilir