GalNAc-T14 may Contribute to Production of Galactose-Deficient Immunoglobulin A1, the Main Autoantigen in IgA Nephropathy.

Introduction: Immunoglobulin A1 (IgA1) with galactose-deficient O-glycans (Gd-IgA1) play a key role in the pathogenesis of IgA nephropathy (IgAN). Mucosal-tissue infections increase IL-6 production and, in patients with IgAN, are often associated with macroscopic hematuria. IgA1-secreting cell lines derived from the circulation of patients with IgAN, compared to those of healthy controls (HCs), produce more IgA1 that has O-glycans with terminal or sialylated N-acetylgalactosamine (GalNAc). GalNAc residues are added to IgA1 hinge region by some of the 20 GalNAc transferases, the O-glycosylation-initiating enzymes. Expression of GALNT2, encoding GalNAc-T2, the main enzyme initiating IgA1 O-glycosylation, is similar in cells derived from patients with IgAN and HCs. In this report, we extend our observations of GALNT14 overexpression in IgA1-producing cell lines from patients with IgAN. Methods: GALNT14 expression was analyzed in peripheral blood mononuclear cells (PBMCs) from patients with IgAN and from HCs. Moreover, the effect of GALNT14 overexpression or knock-down on Gd-IgA1 production in Dakiki cells was assessed. Results: GALNT14 was overexpressed in PBMCs from patients with IgAN. IL-6 increased GALNT14 expression in PBMCs from patients with IgAN and HCs. We used IgA1-producing cell line Dakiki, a previously reported model of Gd-IgA1-producing cells, and showed that overexpression of GalNAc-T14 enhanced galactose deficiency of IgA1, whereas siRNA-mediated GalNAc-T14 knock-down reduced it. GalNAc-T14 was localized in trans-Golgi network, as expected. Conclusions: Overexpression of GALNT14 due to inflammatory signals during mucosal infections may contribute to overproduction of Gd-IgA1 in patients with IgAN.

Galactose-Deficient IgA1 B cells in the Circulation of IgA Nephropathy Patients Carry Preferentially Lambda Light Chains and Mucosal Homing Receptors.

BACKGROUND: IgA nephropathy (IgAN) primary glomerulonephritis is characterized by the deposition of circulating immune complexes composed of polymeric IgA1 molecules with altered O-glycans (Gd-IgA1) and anti-glycan antibodies in the kidney mesangium. The mesangial IgA deposits and serum IgA1 contain predominantly λ light (L) chains, but the nature and origin of such IgA remains enigmatic. METHODS: We analyzed λ L chain expression in peripheral blood B cells of 30 IgAN patients, 30 healthy controls (HCs), and 18 membranous nephropathy patients selected as disease controls (non-IgAN). RESULTS: In comparison to HCs and non-IgAN patients, peripheral blood surface/membrane bound (mb)-Gd-IgA1+ cells from IgAN patients express predominantly λ L chains. In contrast, total mb-IgA+, mb-IgG+, and mb-IgM+ cells were preferentially positive for kappa (κ) L chains, in all analyzed groups. Although minor in comparison to κ L chains, λ L chain subsets of mb-IgG+, mb-IgM+, and mb-IgA+ cells were significantly enriched in IgAN patients in comparison to non-IgAN patients and/or HCs. In contrast to HCs, the peripheral blood of IgAN patients was enriched with λ+ mb-Gd-IgA1+, CCR10+, and CCR9+ cells, which preferentially home to the upper respiratory and digestive tracts. Furthermore, we observed that mb-Gd-IgA1+ cell populations comprise more CD138+ cells and plasmablasts (CD38+) in comparison to total mb-IgA+ cells. CONCLUSIONS: Peripheral blood of IgAN patients is enriched with migratory λ+ mb-Gd-IgA1+ B cells, with the potential to home to mucosal sites where Gd-IgA1 could be produced during local respiratory or digestive tract infections.

Electrocatalytic artificial carbonylation assay for observation of human serum albumin inter-individual properties.

Human serum albumin (HSA) is a multifunctional protein with ligand binding, transporting and buffering properties. Posttranslational modifications and ligand binding processes are closely related to albumin final functional status. In the last few decades, HSA has been characterized using a broad spectrum of methods, but quantitative data on the HSA's modifications among individuals have not been reported. The investigations presented here are based on the non-denaturing electrocatalytic screening of HSA samples isolated from the blood serum of healthy subjects. The electrocatalytic responses of the native protein (Rnat) varied depending on its modifications among individuals, which enable us to express the inter-individual variability. Consequently, the native HSA samples were subjected to ex vivo carbonylation with 50 mM methylglyoxal for 36 h. The differences between Rnat and the responses of artificially carbonylated protein (Rmod) corresponded with inter-individual binding capacity variations (ΔR = Rnat-Rmod). The coefficients of variation for the Rnat and ΔR values of purified HSA samples were estimated to be 8.5 and 23.2%, respectively. A sensitive non-denaturing electrocatalytic assay was utilized to provide new data about albumin inter-individual variations and evaluate its oxidative modifications and binding capacity, which could be used for further studies targeting not only on HSA but also other clinically important proteins.

Total and High Molecular Weight Adiponectin Levels and Prediction of Cardiovascular Risk in Diabetic Patients.

The study aimed at assessing the potential use of lower total and HMW adiponectin levels for predicting cardiovascular risk in patients with type 2 diabetes mellitus (T2DM). Concentrations of total adiponectin or high molecular weight (HMW) adiponectin decrease in association with the development of metabolic dysfunction such as obesity, insulin resistance, or T2DM. Increased adiponectin levels are associated with a lower risk for coronary heart disease. A total of 551 individuals were assessed. The first group comprised metabolically healthy participants (143 females, and 126 males) and the second group were T2DM patients (164 females, and 118 males). Both total adiponectin and HMW adiponectin in diabetic patients were significantly lower when compared with the group of metabolically healthy individuals. There was a weak monotonic correlation between HMW adiponectin levels and triglycerides levels. Binary logistic regression analysis, gender adjusted, showed a higher cardiovascular risk in diabetic persons when both total adiponectin (OR = 1.700) and HMW adiponectin (OR = 2.785) levels were decreased. A decrease in total adiponectin levels as well as a decrease in its HMW adiponectin is associated with a higher cardiovascular risk in individuals with T2DM. This association suggests that adiponectin levels may be potentially used as an epidemiological marker for cardiovascular risk in diabetic patients.

Self care in patients with chronic heart failure. Pilot study - self care includes problems.

AIM: The goal of this pilot study was to define the scope of therapeutic self-care demand in heart failure (HF) patients according to the concepts of self-care postulated by D.E. Orem and to determine the level of problems experienced and self-care in these patients. METHODS: A questionnaire consisting of 7 areas with closed format questions was developed according to the definition. The level of patient problems and level of self-care actions were mapped in each area. The questionnaire was distributed at the hospital outpatient clinic. The study group consisted of 47 heart failure patients (14 women) with following characteristics: average age 68 years, average BMI 29.4, resynchronization therapy 21%, hypertension 69.8%, diabetes mellitus 25.6%, coronary artery disease 46.5%, dilated cardiomyopathy 46.5%, obesity 46.5%, smoking history 39.5% (present and past smoking together), other cardiac disease 16.28%, condition after myocardial infarction 27.8% (NYHA II 41%, NYHA III 56.8%, NYHA IV 2.2%). RESULTS: The greatest problems were in the area of physical activity, sleep and fatigue, the least were in the area of chest pain and blood circulation. The greatest self-care agency was shown in the area of managing problems with physical activities and sleep, the lowest in the area of liquid intake and output. In the patients' subjective opinion, fatigue and sleep problems had the greatest impact on the quality of life. There were no substantial differences in either area based on gender. CONCLUSIONS: The data can serve as a foundation for modifying the extent and structure of patient education for more comprehensive and more effective out­patient treatment of HF.

Procedure for granulocyte collection performed at the Blood Centre of the Faculty Hospital Ostrava.

BACKGROUND: Granulocyte apheresis is a safe and effective method for granulocyte collection. We present a five year experience (2006-2010) of the Blood Center, Faculty Hospital Ostrava, Czech Republic. Donor granulocyte transfusion is one treatment option for haemato-oncology patients with severe neutropenia complicated by bacterial/fungal infections unresponsive to standard antibiotic/antifungal treatment. In this study, we describe the experiences of the Blood Centre at the Faculty Hospital in Ostrava of granulocyte apheresis. METHODS AND RESULTS: A total of 149 granulocyte units were collected for 33 patients from the Department of Haemato-oncology, University Hospital Olomouc, over a 5-year period (2006-2010). Information on donor selection, laboratory screening, mobilization regimen and granulocyte yield was obtained and analyzed. All mandatory screening tests for infection markers, AB0 and Rh and abnormal erythrocyte antibodies were performed. The cytomegalovirus status of the donors was not investigated. Steroids were the only mobilization regimen used, and were generally well accepted. The mean granulocyte yield was 12.6×10(9)/per transfusion unit (range 5.4-30.3). All granulocyte concentrates were irradiated and transfused according to AB0 Rh compatibility within 24 h after collection. CONCLUSION: Based on our experience, granulocytapheresis is a safe and effective method for obtaining granulocytes but the yield can be significantly influenced by other variables. From the recipients' perspective, the use of donor granulocytes supports an effective therapeutic modality.

Haemotherapy analysis: the Teaching Hospital Olomouc in 2006.

AIMS: Annual evaluation of blood transfusion preparation administration at the University Hospital Olomouc, Czech Republic as a contribution to the European Union haemovigilance system. METHODS: Analysis of blood transfusion preparations released from the Department of Blood Transfusion of the above university hospital and laboratory examination results in receivers of these products for the year 2006. Total hospital consumption and usage in particular medical disciplines in the hospital were assessed. RESULTS: Red cell concentrates: in total 14 347 TU (deleucotised in 20.4 %). Departments according to usage: surgery, haematooncology, anaesthesiology and surgery intensive care department, and internal medicine department. Haemoglobin levels were above 100 g/L before blood transfusion administration in 24 % of cases. Platelets: in total 1 712 TU (100 % manufactured by apheresis, 56 % deleucotised). Platelet counts below 20 x 10(9)/L were found in all cases before platelet concentrate administration. Plasma: in total 5 959 TU to 1 297 of patients. Departments according to usage: surgery, anaesthesiology and surgery intensive care department, internal medicine and haematooncology. Two hundred and forty nine cases without coagulation parameter monitoring and 333 (25.67 %) patients with only 1 TU of administered plasma were found. CONCLUSIONS: The incorrect indications for red cell transfusion preparations were found in eleven patients with haemoglobin levels above 130 g/L. Underdosing in 25.67 % of plasma administrations signifies dubious indications in these cases. Implementation of the haemovigilance system in practice is now mandatory for the Czech Republic after joining the European Union. Therefore more precise data and analysis of questionable cases with further education of staff in clinical departments are essential for the haemovigilance principle to be applied on a hospital basis in the Czech Republic.