Inflammatory markers in peripheral arterial disease patients after endovascular revascularization with new restenosis

Wstęp. Zapalenie odgrywa kluczową rolę w powstawaniu i rozwoju miażdżycy tętnic, także u pacjentów z miażdżycą tętnic kończyn dolnych (PAD). Zabiegi rewaskularyzacyjne stanowią czynnik ryzyka, który nasila ten proces, niekiedy prowadząc do powstania restenoz.Cel. Oznaczenie stężenia fibrynogenu, białka C-reaktywnego (CRP), interleukin 6 i 10 (IL-6 i IL-10), zasadowego czynnika wzrostu fibroblastów (bFGF) i transformującego czynnika wzrostu (TGF-b1) we krwichorych z PAD po obwodowej, wewnątrznaczyniowej rewaskularyzacji i u pacjentów z restenozami.Materiał i metody. Przebadano 150 pacjentów z PAD, w tym 90 mężczyzn i 60 kobiet w wieku 50–88 (średnio 65,5) lat. Podczas 12-miesięcznej obserwacji po obwodowej, wewnątrznaczyniowej rewaskularyzacji u 38 pacjentów powstały restenozy. Grupa kontrolna dla oznaczanych cytokin składała się z 15 klinicznie zdrowych osób w wieku 60–83 lat bez niedokrwienia kończyn dolnych. Krew pobierano na czczo do 3,2% cytrynianu sodu w proporcji 9:1 (do oznaczenia fibrynogenu i CRP) i 2,6 ml krwi mieszano z EDTA do pomiaru cytokin: IL-6, IL-10, bFBF, TGF-beta1. Cytokiny badano przy użyciu komercyjnych zestawów metodą immunoenzymatyczną, a fibrynogen i CRP za pomocą analizatora koagulologicznego.Wyniki. We krwi pacjentów z PAD po obwodowej rewaskularyzacji stężenie fibrynogenu i CRP było wyższe od normy laboratoryjnej. Podczas 12-miesięcznej obserwacji parametry te nadal wzrastały: CRP prawie dwukrotnie, a fibrynogen tylko o około 10%. Z badanych cytokin tylko IL-10 i bFGF różniły się istotnie statystycznie od grupy kontrolnej. Cytokiny były również wyższe u pacjentów z powstałymi restenozami, mieściły się jednak, z wyjątkiem TGF-beta1, w szerokich normach laboratoryjnych podanych przez producentów testów.Wnioski. 1. U pacjentów z PAD po wewnątrznaczyniowej rewaskularyzacji stężenie CRP i fibrynogenu podczas rocznej obserwacji wyraźnie się zwiększyło.2. U tych pacjentów obserwowano istotną korelację między prozapalnym CRP i przeciwzapalną interleukiną 10.3. Stężenie CRP, fibrynogenu i cytokin: IL-6, bFGF, TGF-b1 i IL-10 zwiększało się także po powstaniu restenozu pacjentów z PAD, potwierdzając udział w tym procesie miernie nasilonego zapalenia.Introduction. Inflammation plays a central role in the pathophysiology and progression of atherosclerosis, also in patients with peripheral arterial disease (PAD). Revascularization is a risk factor which intensifies this process sometimes leading to newly developed restenosis.Aim. To determine the levels of fibrinogen, C-reactive protein (CRP), interleukin 6 (IL-6), interleukin 10 (IL-10), basic fibroblast growth factor (bFGF) and transforming growth factor-beta 1 (TGF-beta1) in the blood of patients with PAD after endovascular revascularization of lower limbs and in patients with new restenosis.Material and methods. The study included 150 patients with PAD (90 men and 60 women), aged 50–88 (mean 65.5) years after successful peripheral angioplasty (PTA) and/or stenting. Within 12 months after revascularization procedure in 38 PAD patients restenosis occurred. Control group for cytokines consisted of 15 clinically healthy subjects without limb ischaemia aged 60–83 years. Fasting blood was drawn in the morning and put into 3.2% sodium citrate in a 9:1 proportion (for determination of fibrinogen and CRP), and 2.6 ml of blood was mixed with EDTA for determination of cytokines: IL-6, IL-10, bFGF and TGF-beta1. All parameters were measured in the plasma of patients with commercial kits for enzyme immunoassay; fibrinogen and CRP levels were measured with coagulometer.Results. In the plasma of patients with PAD after revascularization, the levels of fibrinogen and CRP were significantly higher than in controls. These markers increased within 12 months of observation: CRP level increased almost two-fold and fibrinogen only by about 10%. All examined cytokines in PAD patients with restenosis were higher than in those without restenosis, but the difference was significant only for IL-10 and bFGF. But increased cytokines in patients with restenosis were within the laboratory norms specified by reagent producers except of TGF-beta1.Conclusions.1.In PAD patients after endovascular revascularization the levels of CRP and fibrinogen during one yearobservation significantly increased.2.In these patients, significant correlation was observed between pro-inflammatory CRP and anti-inflam-matory IL-10.3.The concentrations of CRP, fibrinogen and cytokines (IL-6, bFGF, TGF-beta1 and IL-10) were also higher inPAD patients with restenosis indicating the involvement of low-grade inflammation in this process

Early detection of active glomerular lesions in dogs and cats using podocin

In veterinary medicine, sensitive and specific markers of the early stages of renal failure still remain to be established. Podocytes could be a promising diagnostic tool in veterinary nephrology, especially in the differentiation of active pathological disease and glomerulopathies. Podocin is one of the robust proteins exploitable in detection of podocyturia. This article presents podocyte detection in urine for diagnostic purposes in veterinary medicine using a variety of methods. We describe the advantages and disadvantages of the immunohistochemical technique currently used, and of scanning microscopy, chromatography, and immunostaining. The identification of podocin-positive cells is a promising diagnostic tool in the detection of the early stages of glomerular basement membrane damage. The detection of renal failure prior to the occurrence of azotaemia is of high clinical importance from the clinical and scientific points of view

From Primary MSC Culture of Adipose Tissue to Immortalized Cell Line Producing Cytokines for Potential Use in Regenerative Medicine Therapy or Immunotherapy

For twenty-five years, attempts have been made to use MSCs in the treatment of various diseases due to their regenerative and immunomodulatory properties. However, the results are not satisfactory. Assuming that MSCs can be replaced in some therapies by the active factors they produce, the immortalized MSCs line was established from human adipose tissue (HATMSC1) to produce conditioned media and test its regenerative potential in vitro in terms of possible clinical application. The production of biologically active factors by primary MSCs was lower compared to the HATMSC1 cell line and several factors were produced only by the cell line. It has been shown that an HATMSC1-conditioned medium increases the proliferation of various cell types, augments the adhesion of cells and improves endothelial cell function. It was found that hypoxia during culture resulted in an augmentation in the pro-angiogenic factors production, such as VEGF, IL-8, Angiogenin and MCP-1. The immunomodulatory factors caused an increase in the production of GM-CSF, IL-5, IL-6, MCP-1, RANTES and IL-8. These data suggest that these factors, produced under different culture conditions, could be used for different medical conditions, such as in regenerative medicine, when an increased concentration of pro-angiogenic factors may be beneficial, or in inflammatory diseases with conditioned media with a high concentration of immunomodulatory factors