Prognostic significance of Ki67-negative blast cell clone in the high risk group of children treated for acute myeloid leukaemia.

The aim of this study was to demonstrate the value of immunocytochemical staining of Ki67 antigen expression in blast cells of children with acute myeloid leukemia (AML) and to evaluate its correlation with treatment failure. The material included bone marrow specimens obtained during induction treatment from 46 children treated for AML between 1998-2003. Immunocytochemical staining for Ki67 was based on the ABC technique. Expression of Ki67 antigen on day 0 of induction treatment was confirmed in all patients. The percentage of immunopositive blasts ranged from 88.4% to 99.8% (mean 91.8%). On day 15, according to chemotherapy response, patients were divided into two groups: G1-36 children who responded to induction treatment and reached remission (blast level 5%, low risk group) and G2-10 patients who did not meet remission criterion (blast level > 5%) and were assigned to the high risk (HR) group. Out of 10 children assigned to this group, Ki67 expression in blast cells was confirmed in 4 cases. The fraction of immunopositive blasts ranged from 78.4% to 88.6%. In the other 6 cases, blasts were Ki67-negative. In 12-month period after beginning the treatment, 18 cases of treatment failure (including 7 deceases) were observed in both groups. Five deaths, observed in the HR group, concerned the patients characterized by Ki67-negative blasts. The results indicate a possible correlation between the Ki67-immunonegative blast pattern on day 15 of treatment induction and early decease of AML children assigned to HR group

Nephrotic syndrome unfavorable course correlates with downregulation of podocyte vascular endothelial growth factor receptor (VEGFR)-2

Idiopathic nephrotic syndrome (INS) in children is most commonly caused by primary glomerulopathies. Morphological lesions observed in INS might be secondary to inflammatory factors of mainly extra-renal origin. The vascular endothelial growth factor (VEGF) family is regarded as playing a crucial role in this pathomechanism. The aim of the present work was to analyze the possible relation between VEGF-C and VEGF receptor (VEGFR)-2 expressions at electron microscopy level in different INS cases. The study group comprised 18 children with minimal change disease (MCD), 30 patients diagnosed with diffuse mesangial proliferation (DMP) and 11 subjects with focal segmental glomerulosclerosis (FSGS). An indirect immunohistochemical assay employing monoclonal anti-VEGF-C and anti-VEGFR-2 antibodies was applied in the study. The immunohistochemical expression of VEGF-C within podocyte cytoplasm was significantly increased in DMP subjects who were resistant to steroids and in all FSGS patients compared to MCD children and controls (p < 0.05). VEGF-C over-expression in these cases was followed by downregulation of VEGFR-2. Nephrotic syndrome progression correlates with the downregulation of podocyte VEGFR-2. For this reason, decreased VEGFR-2 expression in the podocyte processes of children with idiopathic nephrotic syndrome might be regarded as a potent factor of unfavorable prognosis. (Folia Histochemica et Cytobiologica 2011; Vol. 49, No. 3, pp. 472&#8211;478

Expression of galectin-3 in nephrotic syndrome glomerulopaties in children.

BACKGROUNDGalectins are a family of ancient animal carbohydrate binding proteins; the name is from their description as beta-galactoside-specific lectins. They have been strongly implicated in inflammation and cancer. Studies of the association of galectins with various aspects of kidney disease in humans are still at an early stage. In line with the above, the aim of the present report was to analyse the immunohistochemical expression of galectin-3 (the only chimera galectin currently identified) in renal biopsy specimens of children with idiopathic nephrotic syndrome (INS).PATIENTS AND METHODSEighteen children with minimal change disease (MCD), 30 with diffuse mesangial proliferation (DMP) and 11 with focal segmental glomerulosclerosis (FSGS) treated between 2003 and 2006 in the Department of Paediatric Cardiology and Nephrology, Poznan University of Medical Sciences. An indirect immunohistochemical protocol using a polyclonal rabbit antibody against human galectin-3 was employed.RESULTSIn the control, MCD and DMP children who responded to steroid therapy anti-galectin-3 reactivity was present both in renal cortex and medulla. It was the strongest within cortical collecting ducts and subjectively less expressed in distal tubules. The total number of galectin-3 positive cortical and medullary segments of collecting ducts was significantly higher in the subjects who did not respond to steroid therapy These patients revealed also immunohistochemical reactivity of galectin-3 within nuclei of individual glomerular mesangial cells (

Neutrophil gelatinase-associated lipocalin and Cathepsin L as early predictors of kidney dysfunction in children with type 1 diabetes

Wstęp: Celem pracy była ocena stężenia w surowicy i w moczu markerów wczesnego uszkodzenia nerek, to jest lipokainy związanej z żelatynazą neutrofilii (odpowiednio sNGAL i uNGAL) oraz wydalania z moczem katepsyny L (uCathL) u dzieci z cukrzycą typu 1 (DM1) wykazujących normoalbuminurię i filtrację kłębuszkową (eGFR) powyżej 90 ml/min/1,73 m2.Materiał i metody: Grupę badaną stanowiło 63 dzieci ze średnim czasem trwania DM1 wynoszącym 5,16 ± 3,39 roku. Albuminurię oceniano za pomocą wskaźnika albuminowo-kreatyninowego (ACR). eGFR obliczano na podstawie stężenia cystatyny C. Hiperfiltrację kłębuszkową (GH) definiowano jako wartość eGFR &gt; 135 ml/min/1.73 m2.Wyniki: U dzieci z DM1 w porównaniu do grupy kontrolnej wykazano znamiennie wyższe stężenia uNGAL, niższe sNGAL i uCathL. Istotne zmiany stężeń uNGAL i uCathL stwierdzono już u dzieci bez GH i z optymalną kontrolą glikemii (HbA1c &lt; 7,5%). Stwierdzono pozytywną zależność pomiędzy uNGAL, ACR i eGFR oraz pomiędzy uCathL i eGFR.Wnioski: Istotne zmiany w stężeniu markerów wczesnego uszkodzenia nerek to jest sNGAL, uNGAL i uCathL mogą wystąpić u dzieci z DM1 i normoalbuminurią. Zmiany uNGAL i uCathL mogą wystąpić nawet u chorych bez GH oraz wykazujących optymalną kontrolę glikemii. Pierwsze objawy zaburzenia czynności nerek w przebiegu DM1 wydają się wynikać z uszkodzenia cewek nerkowych. (Endokrynol Pol 2014; 65 (6): 479–484)Introduction: The aim of this study was to evaluate serum levels and urinary excretion of neutrophil-gelatinase associated lipocalin (respectively sNGAL and uNGAL) and urinary excretion of Cathepsin L (uCathL) in children with type 1 diabetes mellitus (DM1) who presented normoalbuminuria and the estimated glomerular filtration rate (eGFR) above 90 mL/min/1.73 m2.Material and methods: The study group consisted of 63 children with a diabetes duration of 5.16 ± 3.39 years. The degree of albuminuria was based on urine albumin-to-creatinine ratio (ACR), while eGFR was based on serum cystatin C. Glomerular hyperfiltration (GH) was defined as an eGFR value above 135 mL/min/1.73 m2.Results: Children with DM1 showed significantly higher concentrations of uNGAL, and lower sNGAL and uCathL. Significant changes of uNGAL and uCathL levels were even found in children without GH and with optimal glycaemic control (HbA1c &lt; 7.5%). Positive correlations between uNGAL, ACR and eGFR were shown, as well as between uCathL and eGFR.Conclusions: Significant changes in the concentration of markers of early kidney injury: sNGAL, uNGAL, and uCathL, can occur in children with DM1 and normoalbuminuria. The changes of uNGAL and uCathL can be even found in children without GH and with optimal glycaemic control. The earliest signs of diabetic kidney dysfunction seem to result from tubular damage. (Endokrynol Pol 2014; 65 (6): 479–484)

Potentially positive ageing-related variations of medial smooth muscle cells in the saphenous veins used as aortocoronary bypass grafts

Introduction. Currently, elderly people constitute a large proportion of patients undergoing coronary artery bypass grafting (CABG). Activated smooth muscle cells in the tunica media of saphenous vein (SV) grafts are thought to play a key role in the formation of neointima and development of occluding atherosclerotic plaques. The aim of this study was to identify ageing-related variations in the expression of the smooth muscle cells pro­teins that may impact on patency rate of the grafts and the CABG outcomes. Material and methods. The study involved 216 consecutive patients with the mean of 62.7 ± 8.4 years who underwent isolated CABG with at least one SV aortocoronary bypass graft. Expression of a-smooth muscle actin (a-SM actin), smooth muscle-myosin heavy chain (SM-MHC), calponin (CALP), cytokeratin 8 (CK-8), metalloproteinase-2 (MMP-2) and tissue inhibitors of metalloproteinases-2 and -3 (TIMP-2, TIMP-3) in the SV wall was assessed by immunohistochemistry and correlated with the age of patients. Results. Calponin and a-SM actin were expressed in all studied SV transplants. SM-MHC immunoreactivity was observed in SV segments in 68.5% of patients, whereas MMP-2a and TIMPs expression was found in 75% of cases. In more than 50% of analyzed SV transplants, no expression of cytokeratin-8 was found. Moderate correlations between preexisting expressions of either cytoskeletal or hemostatic proteins in the tunica media of the SV grafts and the age of CABG patients were demonstrated. They were positive for SM-MHC (r = 0.494), CALP (r = 0.548), TIMP-2 (r = 0.413) and TIMP-3 (r = 0.406) whereas negative for CK-8 (r = –0.528) and MMP-2 (r = –0.417). Conclusions. Age-dependent decreases in the expression of MMP-2 and CK-8 accompanied by increases in expression of SM-MHC, TIMP-2 and TIMP-3 may promote SV graft patency and, thus, suggest a rationale for common use of SV grafts in the elderly