Prospective Assessment of Hepcidin in Relation to Delayed or Immediate Graft Function in Patients Undergoing Kidney Transplantation.

BACKGROUND: Hepcidin is a peptide hormone that regulates iron homeostasis. Hepcidin may represent an early, predictive biomarker of acute kidney injury, another model of ischemia-reperfusion injury. Urinary hepcidin-25 has been shown to be elevated in patients who do not develop acute kidney injury. Creatinine is an unreliable indicator during acute changes in kidney; therefore, the aim of the study was to assess whether hepcidin could predict renal outcome in 31 consecutive patients undergoing kidney allograft transplantation. Serum hepcidin was evaluated before and after 1, 3, 6, and 10 days after kidney transplantation, using commercially available kits. Serum creatinine was assessed at the same time. METHODS: We found a significant decrease in serum hepcidin, as early as after 1 day after kidney transplantation. Before transplantation, serum hepcidin was related to creatinine. In patients with delayed graft function, there was no decrease in serum hepcidin. RESULTS: Our findings may have important implications for the clinical treatment of patients undergoing kidney transplantation. The "window of opportunity" is narrow in delayed graft function to distinguish between acute rejection and calcineurin inhibitors nephrotoxicity, and time is limited to introduce proper treatment after initiating insult. CONCLUSIONS: Hepcidin must be investigated as a potential early marker for delayed graft function, especially in the upcoming setting of early dialysis treatment or anti-rejection therapy and might contribute to early patient risk stratification.

Diet and the content of selenium and lead in patients with abdominal aortic aneurysm.

BACKGROUND: To evaluate the content of selenium (Se) and lead (Pb) and the influence of dietary habits and smoking in patients with abdominal aortic aneurysm (AAA). PATIENTS AND METHODS: Forty-nine patients with AAA prior to surgical procedures aged 42 - 81 years and a control group of 22 healthy volunteers aged 31 - 72 years and 17 aortic wall samples from deceased were included in the study. Food-frequency questionnaires were implemented in AAA patients to collect the dietary data. Se and Pb concentrations in the serum and blood, respectively, and in arterial wall and parietal thrombus samples were determined by the atomic absorption spectrometry method. RESULTS: The mean Se level in serum of patients with AAA (60.37 ± 21.2 cm/L) was significantly (p < 0.008) lower than in healthy volunteers (75.87 ± 22.4 cm/L). We observed a significant correlation (r = 0.69, p < 0.0001) between the content of Se in serum and the parietal thrombus of examined patients. Se concentration in aortic wall was inversely correlated to the concentration of Pb (r = - 0.38, p < 0.02). We observed significantly lower (p < 0.05) concentrations of Se (39.14 ± 37.1 cm/g) and significantly higher (p < 0.05) concentrations of Pb (202.69 ± 180.6 cm/g) in aortic wall samples of smoking patients than in non-smoking patients (77.56 ± 70.0 cm/g, 73.09 ± 49.8 cm/g; respectively). CONCLUSIONS: Se serum level is lower in patients with AAA than in healthy volunteers. In aortic wall, Se concentration is inversely correlated with Pb concentration. Dietary habits and smoking have an influence on the Se and Pb status in patients with AAA.

Renal parenchyma perfusion spectrum and resistive index (RI) in ultrasound examinations with contrast medium in the early period after kidney transplantation.

The main diagnostic method for renal graft dysfunction is color Doppler ultrasound with the use of spectrae evaluation of blood flow within the main and intrarenal arteries. Ultrasound with contrast medium (US-CM) enhances the possibilities of this tool. The aim of this study was to evaluate the efficacy of US-CM to assess renal graft perfusion among 18 kidney allograft recipients at 5 to 10 days after transplantation. Patients underwent pulse inversion sonography of the graft during intravenous injections of 2.4 mL SonoVue (Bracco-Altana, Italy). Images were quantitatively assessed using computer software to compare the time to peak contrast enhancement effect in the renal cortex and renal pyramids. The results were compared to Doppler ultrasonography of the renal arteries and estimated glomerular filtration rate (eGFR; Modification of Diet in Renal Disease [MDRD]). A correlation was observed between eGFR and blood flow parameters within the renal arteries, as well between the flow time of contrast medium from the artery within the renal hilus to the renal cortex. Increased eGFR correlated with subsequent improvement in graft function (r = -.806; P = .001), and resistive index (RI) of the renal artery was inversely related to subsequent delayed graft function (r = .544; P = .029). Negative correlations were observed between eGFR and renal artery RI, as well as between eGFR and time from renal artery contrast to maximal contrast enhancement of the renal pyramids. A negative correlation was found as well between eGFR and time difference of contrast enhancement of the cortex and pyramids. In conclusion, US-CM enhanced the efficacy of ultrasound diagnostics of the renal graft and may be used as a predictor of graft function in the early posttransplantation period.

Neutrophil gelatinase-associated lipocalin and cystatin C could predict renal outcome in patients undergoing kidney allograft transplantation: a prospective study.

Urinary neutrophil gelatinase-associated lipocalin (NGAL) may represent an early, predictive biomarker of delayed graft function due to ischemia-reperfusion injury. Unfortunately, creatinine is an unreliable indicator of acute changes in kidney function. NGAL was proposed as a novel early marker for detection of acute renal failure. Therefore, the aim of the study was to assess whether NGAL and cystatin C predicted outcomes among 41 consecutive patients undergoing kidney transplantation. Serum NGAL and cystatin C were evaluated before, as well as 1, 3, 6, and 10 days after kidney transplantation using commercially available kits. Serum creatinine was assessed at each time. We observed a significant fall in serum NGAL as early as 1 day following kidney transplantation. Serum cystatin C decreased significantly 3 days after transplantation. Before transplantation, serum NGAL was related to creatinine and cystatin C. At each time point, serum NGAL was related positively to serum creatinine, cystatin C, and negatively to urine volume. In patients with delayed graft function, there was no fall in serum NGAL or cystatin C. Our findings may have important implications for the clinical management of patients undergoing kidney transplantation. The "window of opportunity" to distinguish between acute rejection and calcineurin inhibitor nephrotoxicity is narrow in delayed graft function. Time is limited to introduce proper treatment after the initiating insult. Therefore, NGAL needs to be investigated as a potential early marker for delayed graft function, especially in the settings of early dialysis treatment or antirejection therapy.

Glycosaminoglycan-degrading enzymes in the varicose vein wall.

AIM: Mechanical properties of the vein wall are determined by extracellular matrix components, including glycosaminoglycans (GAGs). The aim of the study was to evaluate the activity of enzymes involved in GAGs degradation pathway in the wall of varicose veins and varicose veins complicated by thrombophlebitis, when compared to the wall of normal ones. METHODS: Normal, varicose veins and varicose veins complicated by thrombophlebitis were collected during surgical treatment of 10 patients. Activities of endoglycosidases, sulphatases and exoglycosidases were assessed according to colorimetric methods. RESULTS: Activities of neutral endoglycosidases degrading chondroitin-4-sulphate (C4S) and heparan sulphate (HS) were decreased, whereas activities of neutral endoglycosidases degrading dermatan sulphate and hyaluronic acid were increased in varicose veins and varicose veins complicated by thrombophlebitis. Activities of acidic endoglycosidases degrading C4S and HS were decreased in varicose veins and varicose veins complicated by thrombophlebitis, whereas activity of acidic endoglycosidases degrading chondroitin-6-sulphate was decreased only in varicose veins complicated by thrombophlebitis. Furthermore increased activities of arylosulphatase B, beta-N-acetylhexosaminidase and alpha-L-iduronidase were demonstrated in varicose veins, as well as in varicose veins complicated by thrombophlebitis. CONCLUSIONS: Changed activities of GAGs-degrading enzymes may contribute to previously reported changes in the content and molecular differentiation of GAGs in the wall of varicose veins that may play a role in the disease pathogenesis.

The predictive value of arterial renal blood flow parameters in renal graft survival.

BACKGROUND: Renal allograft survival depends on a number of factors, however, no reliable simple parameter has been shown to predict long-term outcome after transplantation. Ultrasound is recognized and relatively inexpensive, providing information about renal location, contour, and size. Doppler ultrasonography shows kidney morphology and hemodynamics. The aim of this study was the evaluation of whether Doppler ultrasound of renal arteries performed in the early stage after transplantation was a valuable predictor for long-term-outcomes. MATERIAL AND METHODS: The study included 17 female and 24 male patients, aged 17-69 years with stable graft function. The Doppler ultrasound of renal flow was done on the 1st and 3rd day after transplantaion, and estimated glomerular filtration rate (eGFR) on the 20th day. The measured indices were as follows: maximum blood flow velocity (V(max)), minimum blood flow velocity (V(min)), resistive index (RI), and pulsatile index (PI). The creatinine concentration was evaluated, and eGFR calculated. RESULTS: Mean renal and intrarenal artery RI increased to day 3 after transplantation, and then reduced. The mean renal and intrarenal V(max) at day 3 correlated positively with eGFR (r = 0.38; P = .015); (r = 0.45; P = .003, respectively). Mean renal and intrarenal V(min) correlated positively with eGFR (r = 0.50; P = .001; r = 0.41; P = .008, respectively). The mean renal and intrarenal V(max) and V(min) on day 1 did not correlate with eGFR. CONCLUSIONS: Early Doppler Ultrasonography of renal graft hemodynamics may be a valuable predictor of graft survival and long-term outcomes. Blood flow velocity within renal arteries seemed to be an important factor.

Renal artery resistance index, thyroid hormones, and thyroid volume in the early kidney transplants recipients.

BACKGROUND: Thyroid hormones could affect renal function, and, on the other hand, renal dysfunction may affect thyroid function. Disturbances of concentrations of thyroid hormones are often associated with thyroid gland enlargement. The aim of the study was to assess the function and morphology of the thyroid (volume and hormones concentration) and kidney function after transplantation (creatinine concentration and resistance index [RI] of transplant artery). MATERIAL AND METHODS: The group included 13 females, 19 males; aged 19-69 years, mean 44.75 +/- 14.8 years after transplantation with stable graft function. Thyroid volume, renal artery RI, creatinine concentration, and concentrations of T3, rT3, FT3, FT4, and TSH were estimated the day before surgery, and at 1, 3, 6, and 10 days after transplantation. RESULTS: The statistical analysis revealed a negative correlation between delta RI (difference between RI at 3 and 6 days after transplantation) and serum creatinine concentration, 10 days after transplantation (r = -0.63; P < 0.01). We also observed a negative correlation between creatinine serum concentration at 10 days after transplantation and delta thyroid volume (Delta Vol; r = - 0.48; p < .05), a positive correlation between delta FT4 (Delta FT4) serum concentration, and delta creatinine (Delta Crea; r = 0.73; P < .001). CONCLUSIONS: The dynamics of RI changes in the transplant kidney artery between 3 and 6 days after transplantation may predict graft function. Together with improved kidney function at 10 days after transplantation, we observed a regression of goiter.

The effect of temperature, acidification, and alkalization changes as well as ethanol on salivary cathepsin D activity.

The activity of salivary cathepsin D undergoes inactivation at the temperature of 50-60 degrees C and at pH of 2.0 and pH of 8.0-10.0. The enzyme activity is also decreased by high concentrations of ethanol and high-proof alcoholic beverages. The factors should be taken into consideration in the evaluation of salivary cathepsin D activity.

The choice of conditions for cathepsin D activity determination in human saliva.

The aim of the study was the demonstration and choice of conditions for the determination of cathepsin D activity in human mixed saliva. The 6% solution of hemoglobin, denatured with hydrochloric acid, was used as the substrate. The ratio of saliva volume to hemoglobin was 4:1 w/v. The reaction was interrupted by adding 10% trichloroacetic acid, after 6 hours of incubation at 37 degrees C. The increase in degradation products was determined with the use of Folina and Ciocalteau method with copper modification.

Activity of coagulation and fibrinolytic system components in the vein thrombus.

PURPOSE: Behaviour of the vein thrombus is determined by the activity ratio of coagulation factors to factors of fibrinolytic system. The aim of the study is to evaluate activity of some coagulation and fibrinolytic factors in the vein thrombus. MATERIAL AND METHODS: The activity of platelets aggregating factors, tissue factor, thrombin, antithrombins, antiheparin factors, plasminogen activators, plasmin (plasminogen) and antiplasmins of the vein thrombus homogenate was determined using coagulative, fibrinolytic and caseinolytic tests. Retracted blood clot was a compared material. RESULTS: Tissue factor activity in the vein thrombus was above twofold higher and antiheparin activity was nearly twice higher in comparison to the blood clot. The vein thrombus contains also active thrombin. Plasminogen activators activity in the vein thrombus was twofold higher and activity of plasmin (plasminogen) was threefold higher than in the blood clot. High activity of the tissue factor, substances neutralizing heparin and presence of thrombin intensify the thrombus enlargement. However, the thrombotic tendency may be balanced by a high activity of plasminogen activators and high activity of plasmin (plasminogen). CONCLUSIONS: 1) Vein thrombus is characterized by high activity of tissue factor, presence of active thrombin and high antiheparin activity. 2) High coagulative potential of vein thrombus is balanced to a certain grade by high fibrinolytic potential: high activity of plasminogen activators and high activity of plasmin (plasminogen), as well as absence of antiplasmins activity.

Matrix metalloproteinases in the vein wall.

AIM: Matrix metalloproteinases contribute to extracellular matrix remodelling that can influence mechanical properties of the vein wall and predispose to varicose veins development. The aim of the study was to assess the following matrix metalloproteinases in the wall of varicose veins: tissue collagenase I (MMP-1), gelatinase A (MMP-2), gelatinase B (MMP-9) and stromelysin 1 (MMP-3). METHODS: Normal, varicose and varicose veins complicated by thrombophlebitis were collected during the surgical treatment of 26 patients. In harvested tissues the presence of gelatinases was detected with zymography, contents of MMP-1, MMP-2, MMP-3 and MMP-9 were evaluated with ELISA, activity of MMP-1 was assessed with HPLC and activity of MMP-2 with ELISA. RESULTS: Zymography demonstrated particularly high contents of both gelatinases in the wall of varicose veins complicated by thrombophlebitis. The contents of MMP-1, MMP-2 and MMP-9 were significantly increased only in the wall of varicose veins complicated by thrombophlebitis, whereas the increased content of MMP-3 was also found in the wall of varicose veins. A significantly higher activity of MMP-1 was shown only in the wall of varicose veins complicated by thrombophlebitis, whereas an active form of MMP-2 was increased in the wall of varicose, as well as varicose veins complicated by thrombophlebitis, when compared with normal ones. CONCLUSION: The wall of varicose veins, particularly those complicated by thrombophlebitis shows extensive alterations in the content and activity of matrix metalloproteinases, that may result in extracellular matrix remodelling, influence mechanical properties of the vein wall and predispose to further progression of the disease.

Amidolytic activity of plasma euglobulins.

Amidolytic activity of plasmin, produced in euglobulin fraction, does not correlate with the time of euglobulin fibrinolysis. It does not depend on fibrinogen concentration.

Chemical compound content and enzyme activity in supernatant and sediment of liver homogenate.

Evaluation was performed of chemical compound contents and enzyme activities in the whole homogenate, its supernatant and sediment. Six rabbit livers were pulverized in liquid nitrogen and homogenized. After centrifugation, the contents of protein, haemoglobin, vitamin A, vitamin E, vitamin C, as well as the activities of cathepsin B, cathepsin D, superoxide dismutase, catalase, glutathione peroxidase and reductase were assessed in the whole homogenate, its supernatant and sediment. Protein, vitamin A, superoxide dismutase, catalase, cathepsin D, glutathione peroxidase and reductase reveal uniform localisation. Vitamin C and cathepsin B are localized in supernatant, whereas haemoglobin is localized mainly in sediment. Evaluation of chemical compounds and enzyme activities should be performed in the whole homogenate, supernatant and sediment to obtain a real interpretation of biochemical disturbances in the investigated material.

Assessment of connective tissue fibres in walls of allogenic arterial grafts preserved by the method of cold ischemia--a preliminary report.

The aim of the study was to evaluate microscopic changes in the wall structures of allogenic arterial grafts, preserved by the method of cold ischemia in relation to the storage period and to test the possibility of the storage period prolongation by allograft freezing at -70 degrees C. The middle layer ultrastructure is well preserved till 30 days from allograft harvesting, however, allograft freezing results in total destruction of elastic and collagen fibres in the arterial wall. An application of allogenic arterial grafts, preserved by the method of cold ischemia till 30 days from their harvesting, seems an efficient therapeutic method in the treatment of patients with synthetic vascular graft infection. Further prolongation of the storage period at -70 degrees C made the allograft useless for implantation.

Some components of oxidative-antioxidative system in human blood plasma and serum.

Comparison of the concentrations and activities of components in the oxidative-antioxidative system between blood plasma and serum. Blood plasma and serum samples were obtained from 38 healthy adults to evaluate malondialdehyde concentration, the total antioxidative capacity, superoxide dismutase activity, protein and non-protein sulphydryl groups, ascorbate, haemoglobin, methaemoglobin and protein. Blood plasma shows higher activity of superoxide dismutase, as well as higher concentrations of low-molecular sulphydryl groups and ascorbate, when compared to those in blood serum. The total plasma antioxidative capacity is also higher than that assessed in blood serum. Processes of blood coagulation and blood clot retraction lead to antioxidant consumption. The evaluation of oxidative-antioxidative system for diagnostic purposes should be performed in blood plasma.

Haemoglobin of varicose vein, varicose vein with thrombophlebitis and in parietal thrombus of varicose vein.

Saphenous veins were taken for examination: unchanged, varicose with thrombophlebitis and varicose thrombus. The contents of haemoglobin and protein were determined in the homogenate of that material. Only small quantities of haemoglobin were found in walls of unchanged veins. Greater amounts of haemoglobin were observed in walls of varicose veins, especially in walls of varicose veins with thrombophlebitis. The varicose vein thrombus also contained marked quantities of haemoglobin.

[Proteolytic activity of placenta with EPH-gestosis determined by casein and azocasein]. / Aktywnosc proteolityczna lozysk EPH-gestozowych oznaczana przy uzyciu kazeiny i azokazeiny.

Total proteolytic activity, activity of cathepsin B, activity of cysteine cathepsins and contents of protein degradation products were determined in placentas of pregnancies complicated with mild, moderate and severe EPH-gestosis and in placentas from normal pregnancies. The highest activity of all the determined proteases was observed in placentas of pregnancies complicated with severe EPH-gestosis. The placentas of pregnancies complicated with severe EPH-gestosis also include the highest amounts of aminoacids and low-molecular peptides.

[Activation mechanisms, biological role and inhibitors of metalloproteases in the extracellular matrix]. / Mechanizmy aktywacji, znaczenie biologiczne i inhibitory metaloproteaz macierzy miedzykomórkowej.

Prometalloproteinases are activated by serine proteases, MMP-3, leucocytic elastase, furin, furin-like proteases and by membrane-type metalloproteinases as well. They form complexes with some proMMPs and thus they modify their activation.

[Activity of cathepsin B and cystatins in the placenta during EPH-gestosis]. / Aktywnosc katepsyny B i aktywnosc cystatyn w lozysku EPH-gestozowym.

Activity of cathepsin B using Bz-DL-Arg-pNA contents of SH-group by means of Ellman method, activity of cystatins against papain using casein as a substrate and contents of deoxyribonuclein acids by Burton method were determined in 64 placentas of pregnancies with EPH-gestosis and in 36 placentas of physiological pregnancies. The placentas from pregnancies with EPH-gestosis showed markedly higher activity of cathepsin B, no difference in the contents of SH-group, slightly higher activity of cystatins and they contain less deoxyribonucleic acids than the placentas from physiological pregnancies. The obtained results show that proteolytic--anti-proteolytic balance in placentas from pregnacies with EPH-gestosis is changed to the advantage of cathepsin B. This protease may in formation of structural and functional changes observed in placentas during EPH-gestosis.

[Experimental aortic aneurysm]. / Eksperymentalny tetniak aorty.

Experimental aortic aneurysm may be evoked in animals by application of chemical compounds disturbing biosynthesis, posttranslating modifications and degradation of elastin and collagen, local damage of the aortic wall by chemical, thermal and mechanical factors and by increased blood pressure. Administration of protease inhibitors, anti-inflammatory drugs, agents enhancing synthesis and formation of elastic and collagen fibers and hypotensive drugs prevent formation enlargement and rupture of aneurysm.