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

Pre‐pubertal treatment with a Gn RH agonist in bitches—Effect on the uterus and hormone receptor expression

ContentsAim of the study was to examine the effect of deslorelin on uterine tissues of eleven pre‐pubertal bitches aged 4.2 ± 0.6 m. Implants containing placebo (sodium chloride 0.9%; n = 4, G I), 4.7 mg (n = 3, GII) or 9.4 mg (n = 4, GIII) deslorelin acetate (Suprelorin®; Virbac, France), were administered subcutaneously. Signs of oestrus, vaginal cytology, serum progesterone (P4) and estradiol‐17β (E2) concentrations were monitored until the occurrence of oestrus. Bitches were ovariohysterectomized and sections from the uterine tissue were subjected to immunohistochemistry (IHC) for detection of GnRH receptor (R), Kisspeptin (KP)10, Kisspeptin receptor (GPR54), androgen receptor (AR), oestrogen receptor (ER) α,β, and progesterone receptor (PR). Tissue sections were scored semi‐quantitatively using an immunoreactivity score (IRS) ranging from 0 to 300 (3). Since some animals were ovariohysterectomized before puberty (n = 1 from GII and n = 2 from GIII), and some in metestrus (all controls and 2 from GII and GIII each), results from these animals were separately evaluated and compared to the controls. Results: No abnormalities were seen in uterine tissues. Kisspeptin 10 expression was low in all cell types, highest IRS were seen in the vascular endothelial cells. The GPR54 was mainly detected in the luminal epithelial cells, superficial and deep uterine glands. The expression of GPR54 and ERα,β was especially high in bitches operated prepubertally. No difference was observed between the controls and experimental bitches operated in their first metestrus. The PR and ERα,β were exclusively expressed in superficial and deep uterine glands and luminal surface epithelial cells. The AR and GnRH‐R expression was negative in all cells of all groups. We conclude that application of 4.7 or 9.4 mg deslorelin at the age of 4 months did not cause uterine disturbances. GPR54 expression might be influenced by pre‐pubertal deslorelin treatment or the changings related to approaching puberty; the latter is supposed in case of ERα,β.</jats:p