Serum PCSK9 Correlates with PTX3 and Apolipoproteins B, A1, and C3 Concentrations in Patients with Type 2 Diabetes.

Proprotein convertase subtilisin/kexin type 9 (PCSK9) is involved in the regulation of LDL metabolism. There is evidence that circulating PCSK9 is a cardiovascular risk factor. In this study, we determined factors associated with circulating PCSK9 in a group of patients with type 2 diabetes mellitus (DM2). Material included 116 consecutive patients with DM2 from outpatient diabetes clinic. Circulating PCSK9, PTX3, apolipoprotein (apo) B100, apo B48, and apo C3 levels were determined by ELISA, apo A1 by immunoturbidimetry. The mean (sd) age of patients was 59.1 (11.1) years, the mean (sd) values of serum PCSK9 were 255.4 (106.97) ng/ml. Circulating PCSK9 correlated negatively with age (r = -0.21, p < 0.05) and HbA1c (r = -0.21, p < 0.05) and positively with BMI (r = 0.21, p < 0.05), total cholesterol (r = 0.59), LDL-cholesterol (r = 0.50), triglyceride (r = 0.35), apo B100 (r = 0.43), apo A1 (r = 0.43) (p < 0.001 for all), apo C3 (r = 0.29, p < 0.01), and apo B48 (r = 0.25, p < 0.01) concentration and FLI (r = 0.26, p < 0.01). Strong correlation between PTX3 and PCSK9 levels was observed (r = 0.47, p < 0.001). Multiple stepwise backward regression analysis with PCSK9 as dependent variable revealed that PTX3, apo B100, apo A1, apo B48, and BMI were significantly positive and the presence of NAFLD and HbA1c negatively associated with PCSK9 concentrations. These variables together explain 57% of PCSK9 variability; the strongest relationship was observed between PCSK9 and PTX3 and apo B100. Our results indicate that circulating PCSK9 is significantly associated with inflammation marker PTX3 as well as atherogenic lipids and apolipoproteins C3, B100, and B48, which might be of value in understanding interactions between development of atherosclerosis and inflammatory state in DM2 patients.

Correlates of pentraxin 3 serum concentration in men and women with type 2 diabetes.

Elevated levels of plasma pentraxin 3 (PTX3), a marker of inflammation, are associated with the risk of developing cardiovascular diseases in the general population, as well as in patients with type 2 diabetes (DM2). In this study, we aimed to determine factors associated with PTX3 serum concentrations in men and women with DM2. The study included 116 consecutive patients (67 men and 49 women) with DM2 from an outpatient diabetic clinic. Men were characterised by lower age and higher uric acid, creatinine and bilirubin concentrations and waist/hip ratio than women. In women, low-density lipoprotein cholesterol (LDL-C) levels were higher than in men. In men, median (interquartile range) values of PTX3 concentration were 4.02 (1.99), and in women they were 4.53 (3.31) ng/ml (NS). In men, PTX3 concentrations correlated with total cholesterol (TC), triglycerides, apolipoprotein (Apo) C3, Apo B48, Glc and creatinine levels. In women, PTX3 correlated significantly with TC and LDL-C and Apo B100. Partial regression analysis revealed that after adjusting for age, PTX3 concentrations in men were significantly associated with TC, LDL-C, triglycerides, creatinine, Apo C3 and Apo B48, while in women they were associated with TC, LDL-C and Apo B100. The results could be of importance in sex-specific prevention of vascular complications in DM2 patients.

Serum pentraxin 3 concentration in patients with type 2 diabetes and nonalcoholic fatty liver disease.

INTRODUCTION: Nonalcoholic fatty liver disease (NAFLD) is common in patients with type 2 diabetes (T2D). Pentraxin 3 (PTX3), a marker of inflammation, is a cardiovascular risk factor. OBJECTIVES: We examined clinical and biochemical factors associated with serum PTX3 concentrations in patients with T2D with and without NAFLD. PATIENTS AND METHODS: Serum material was obtained from 116 patients with T2D (mean age, 59.1 years), including 79 patients with NAFLD. RESULTS: Median (interquartile range) PTX3 level was 4.264 (2.293) ng/ml in patients with and 3.773 (3.223) ng/ml in patients without NAFLD (P = 0.93). In the whole group, PTX3 level was associated with total cholesterol, low­density lipoprotein cholesterol (LDL­C), apolipoprotein (apo) B100, apo C3, triglyceride (TG) concentrations, and waist circumference after adjustment for age and gender. As indicated by partial regression coefficient b, increase of independent variable LDL­C by 1 mmol/l was associated with the rise of PTX3 by 1.2017 ng/ml, increase of apo B100 by 1 mg/dl with the rise of PTX3 by 1.0051 ng/ml, and increase of apo C3 by 1 µg/dl with the rise of PTX3 by 1.0012 ng/ml. In patients with T2D with NAFLD, total cholesterol, LDL­C, TG, apo C3, and apo B100 were associated with PTX3. Associations of PTX3 with apolipoproteins were observed only in the NAFLD group. CONCLUSIONS: Reported associations of PTX3 level add new insight into possible mechanisms of its atherogenic actions.

Development of sampling method and chromatographic analysis of volatile organic compounds emitted from human skin.

AIM: The studies on volatile organic compounds emitted from skin are an interest for chemists, biologists and physicians due to their role in development of different scientific areas, including medical diagnostics, forensic medicine and the perfume design. This paper presents a proposal of two sampling methods applied to skin odor collection: the first one uses a bag of cellulose film, the second one, using cellulose sachets filled with active carbon. MATERIALS & METHODS: Volatile organic compounds were adsorbed on carbon sorbent, removed via thermal desorption and analyzed using gas chromatograph with mass spectrometer. RESULTS: The first sampling method allowed identification of more compounds (52) comparing to the second one (30). Quantitative analyses for acetone, butanal, pentanal and hexanal were done. CONCLUSION: The skin odor sampling method using a bag of cellulose film, allowed the identification of many more compounds when compared with the method using a sachet filled with active carbon.

Serum uromodulin concentrations correlate with glomerular filtration rate in patients with chronic kidney disease.

INTRODUCTION Urinary uromodulin excretion has been associated with kidney diseases. However, serum uromodulin concentrations have not been extensively studied in patients with chronic kidney disease (CKD), and the results of published studies are inconsistent. OBJECTIVES The aims of the study were to evaluate serum uromodulin concentrations in patients with CKD and to assess the utility of serum uromodulin measurements for diagnosing CKD stages. PATIENTS AND METHODS This observational study included 170 patients with CKD stages 1 to 5, not treated by renal replacement therapy, and 30 healthy individuals. The serum levels of creatinine, cystatin C, and uromodulin were measured, and estimated glomerular filtration rate (eGFR) was calculated according to the 2012 CKD Epidemiology Collaboration cystatin­creatinine equation. RESULTS Among patients with CKD, serum uromodulin concentrations were significantly lower than in controls, and were strongly negatively correlated with renal retention markers (ie, serum creatinine and cystatin C) and strongly positively correlated with eGFR. An inverse, hyperbolic relationship between serum creatinine and uromodulin levels was analogous to the well­known association between serum creatinine concentrations and eGFR. A receiver­operating characteristic curve analysis showed a high diagnostic accuracy of the measurement of serum uromodulin concentrations in the assessment of CKD stages. CONCLUSIONS Serum uromodulin concentrations are closely correlated with eGFR, which is the recommended measure of renal function. As uromodulin is produced exclusively by renal tubular cells, the assessment of uromodulin levels in patients with CKD may be an alternative method for evaluating the number of functioning nephrons.

[Patient receiving peritoneal dialysis after treatment of ovarian cancer]. / Pacjentka dializowana otrzewnowo, po leczeniu nowotworu zlosliwego jajnika.

Peritoneal dialysis is one of the three available options for renal replacement therapy. This method of treatment of end-stage renal disease gives patients relatively high sense of independence and control over their disease, especially in comparison with hemodialysis, and therefore is often preferable method for young individuals wishing to lead an active lifestyle. We present a case of 22 year old female patient with stage 5 of chronic kidney disease, which is a consequence of multi-agent chemotherapy for endo-dermal sinus tumor of the right ovary (diagnosed at the age of 13). Particularly important in the context of treating our patient with peritoneal dialysis is the fact of confirmed metastases into the peritoneum, which was the reason for the use of chemotherapy reserved for high-risk patients (ifosfamide + etoposide + cisplatin). The selected program of chemotherapy provided effective eradication of cancer, but a side effect of treatment was renal tubular damage. In the period from 03.2006 to 05.2007 our patient required hemodialysis (with gradually reduce dose of dialysis), at a later time to 12.2011 patient did not require renal replacement therapy (stable renal function were observed at the stage 4 of chronic kidney disease), but in 12.2011 resumption of dialysis was necessary and the patient, in accordance with her selection, is receiving peritoneal dialysis. Qualification of our patient for treatment with peritoneal dialysis was associated with reasonable concern about the ability to provide acceptable adequacy of dialysis. Apprehensions proved to be unfounded, the clinical observation of the patient presents proper ratios of dialysis adequacy. Our patient was also qualified for renal transplantation.

Peritoneal solute transport rate as an independent risk factor for total and cardiovascular mortality in a population of peritoneal dialysis patients.

The aim of the present study was to assess the influence of peritoneal permeability expressed as the dialysate-to-plasma ratio of creatinine (D/P Cr) on total and cardiovascular (CV) mortality in a population of peritoneal dialysis (PD) patients during a 6-year observation period. The study recruited 55 patients (mean age: 53 years) treated with PD for a median of 24 months. Hematology parameters and serum albumin were determined using routine methods. Tumor necrosis factor alpha (TNF-alpha) and transforming growth factor beta (TGF-beta1) were determined by high-sensitivity ELISA. Peritoneal transport characteristics were identified using D/P Cr reference values after a peritoneal equilibration test. During the 6-year observation period, 22 patients (40%) died, mostly from CV complications (77% of deaths). In multiple Cox regression, D/P Cr and dialysate volume at PD initiation predicted total [hazard ratio (HR): 1.57; p = 0.02; and HR: 1.20; p = 0.04 respectively] and CV mortality (HR: 1.65; p = 0.02; and HR: 1.23; p = 0.05 respectively) independent of age, dialysis therapy duration, serum albumin concentration, dialysis adequacy measures, TGF-beta1, and TNF-alpha. Additionally, TNF-alpha was independently associated with all-cause and CV mortality, and albumin, with all-cause mortality. Baseline D/P Cr was a strong independent marker of survival in PD patients. Baseline D/P Cr and dialysate volume were independent risk factors for total and CV mortality in the PD population and could be significant for assessing CV risk in this population.

Detection of potential chronic kidney disease markers in breath using gas chromatography with mass-spectral detection coupled with thermal desorption method.

The analytical potential of chromatographic breath analysis towards detection of compounds suggested as markers of chronic kidney disease (CKD) was tested. Until now, trimethylamine (TMA) considered as a potential marker of renal disorder was detected mainly in plasma. Detection of TMA in breath was rarely undertaken due to analytical difficulties associated with amines' properties. The results of our investigations confirmed that an application of thermal desorption (TD) and gas chromatography with mass-spectral detection (GC/MS) allows direct detection of TMA in breath. The preliminary studies allowed to determine the breath composition in case of patients suffering from CKD and to compare the obtained results to a control group. Breath samples were collected from 14 patients and 9 healthy volunteers. TMA was detected in all patients suffering from CKD in the range 1.76-38.02ppb, but not in the control group. Acetone and isoprene were present in the exhaled air of all examined persons. The concentration of acetone was in the range of 26.52-329.46ppb in the patient group and 73.11-437.14ppb in the control group. Isoprene was detected in the range 57.17-329.8ppb among CKD patients and 27.99-143.77ppb in healthy volunteers. Additionally aliphatic hydrocarbons and sulfur compounds were determined in breath as compounds which could be essential in case of diseases coexisting with CKD. Apart from TMA and pentane no statistically significant differences were found using our analytical technique. TMA was detected in the breath of all patients with CKD and in none of breath samples in control group. TMA seems to be a promising marker of CKD.

Intima-media thickness correlates with features of metabolic syndrome in young people with a clinical diagnosis of familial hypercholesterolaemia.

BACKGROUND: Familial hypercholesterolaemia (FH) is a monogenic lipid metabolism disorder characterised by markedly elevated serum low-density lipoprotein (LDL) cholesterol level due to a mutation in the LDL receptor gene. Clinical features of FH include premature atherosclerosis and coronary artery disease. AIM: To explore associations between noninvasive markers of atherosclerosis including intima-media thickness (IMT) and pulse wave velocity (PWV) and blood lipids, blood pressure (BP) and obesity in a group of young patients with FH. METHODS: Study population included 36 patients aged < 35 years with the diagnosis of FH based on the Simon Broome Register criteria, and their 49 relatives who comprised the control group free of FH. RESULTS: Mean IMT values were higher in FH patients than controls (0.60 ± 0.19 vs. 0.53 ± 0.07 mm, respectively, p < 0.05).Mean body mass index (BMI) and waist circumference were similar in patients and controls. The prevalence of carotid atherosclerotic plaques was significantly higher among FH patients (n = 6) than in controls (n = 1) (21.4% vs. 2.6%, p = 0.012). Arterial hypertension was present in 27.8% of patients with FH and 16.3% of subjects in the control group. Systolic blood pressure (SBP) in FH patients correlated significantly with age (r = 0.35), BMI (r = 0.48) and waist circumference (r = 0.47), and diastolic blood pressure (DBP) correlated with BMI (r = 0.42) and waist circumference (r = 0.41). PWV correlated significantly with age (r = 0.44), SBP (r = 0.63) and DBP (r = 0.52). We did not find any correlations between IMT and serum lipids, BP or obesity indices in FH patients. CONCLUSIONS: Our findings show a higher rate of arterial hypertension in young FH patients compared to their relatives free of FH, with significant associations between BP and indices of obesity and arterial stiffness. Intensive lipid lowering and antihypertensive therapy along with a reduction in central fat may be considered a mandatory treatment strategy in young FH patients to prevent atherosclerosis and increased arterial stiffness.

[New opportunities of renal diseases diagnostics using gas chromatography]. / Nowe mozliwosci diagnostyki chorób nerek przy wykorzystaniu chromatografii gazowej.

During the last decade the amount of patients suffering from chronic kidney disease (CKD) has increased. The physicians' efforts are focused on early CKD diagnosis and reduction of the end-stage renal diseases incidence. The breath test seems to be a promising diagnostic device offering early noninvasive diseases detection. The aim of presented study was the determination of breath composition in case of persons suffering from CKD. Breath samples were collected from 14 patients and 7 healthy volunteers. Exhaled air samples were analyzed by gas chromatography equipped with mass spectrometer (GC-MS). Samples were enriched using solid phase microextraction (SPME). Trimethylamine (TMA), mentioned in literature as potential marker of chronic kidney diseases, was detected in case of 11 patients. Among breath components were detected: sulfur compounds: dimethyl sulfide (was detected in exhaled air of patients and control group), carbon disulfide (was detected in case of 4 healthy and in case of all patients) and also potential markers of oxidative stress: propane, butane, pentane, 2-methylpentane, hexane. Acetone and isoprene occurred in exhaled air of all studied persons. The considerable increase of acetone concentration in comparison to control group was observed in case of patients with diagnosed diabetes. The application of gas chromatography with mass spectrometer and appropriate enrichment of samples allows to define the breath profile characteristic for chosen the unit of disease. Typical compounds--biomarkers can be useful for early diagnostics.

A novel mutation (Cys308Phe) of the LDL receptor gene in families from the South-Eastern part of Poland.

The purpose of this investigation was to characterize a new mutation in the LDL-receptor (LDLR) gene in three families with clinically diagnosed familial hypercholesterolemia (FH) from the South-Eastern part of Poland. Mutational screening with exon by exon sequencing analysis was performed in all probands. The novel mutation c986G>T (Cys308Phe) in the exon 7 of LDLR gene was found in three apparently unrelated probands with FH. Analysis of the receptor activity of peripheral blood lymphocytes by binding and uptake of DiL-LDL showed a significant reduction (by 24% versus healthy control) of the fluorescent label in the lymphocytes of patients heterozygous for this mutation. Concentrations of serum LDL-C in probands before treatment were between 9.5 and 10.5 mmol/l. All patients had corneal arcus and tendon xanthoma. Clinically, families were characterized by premature coronary artery disease. This mutation occurred relatively frequently in our group of patients with FH, but this could be explained by a founder effect since we demonstrated their common ancestors.

Corticosteroid treatment of kidney disease in a patient with familial lecithin-cholesterol acyltransferase deficiency.

Familial lecithin-cholesterol acyltransferase (LCAT) deficiency (FLD) is a rare genetic disorder of lipid metabolism, characterised by low plasma HDL cholesterol, proteinuria, haemolytic anaemia and corneal opacities. Usually renal disease progresses during the third decade of life to renal failure; however the pathogenesis of renal disease is not well understood. In this study we describe treatment of renal disease in two siblings with FLD. The proband WX at the age of 31 years presented proteinuria and ankle oedema during her third pregnancy. Diagnosis of FLD was based on a renal biopsy with characteristic serpiginous fibrillar deposits under electron microscopy, markedly decreased HDL cholesterol, esterified cholesterol levels and LCAT activity, confirmed by molecular analysis. After 3 years her proteinuria increased and she received an ACE inhibitor to which she responded well. During further increases of proteinuria she additionally received methylprednisolone and her proteinuria decreased. This long-term observation indicates the efficacy of corticosteroids and renin-angiotensin-aldosterone system blockers in the treatment of proteinuria in patients with FLD. The results suggest the role of inflammatory processes as well as dyslipidemia in the pathogenesis of glomerular disorders in LCAT-deficient patients.

Does coronary artery calcification in patients with diabetic nephropathy depend on the advancement of renal failure?

Calcium-phosphate disorders and vascular calcification are highly prevalent in patients with diabetes mellitus and nephropathy. The aim of the study was to compare the prevalence and advancement of vascular calcification in patients with end-stage diabetic nephropathy on peritoneal dialysis and diabetic patients with chronic renal disease stages 2-4. The study group included 31 patients with type 2 diabetes and diabetic nephropathy divided into 2 groups: 12 patients (aged 50-74 years: mean 58.6+/-8.8) undergoing peritoneal dialysis and 19 patients (aged 46-82 years; mean 65.8+/-9.7) with chronic kidney disease stages 2-4 (GFR range 24-78 ml/min/1.73 m2). Coronary artery calcification score, was assessed using multi-slice computed tomography. Coronary artery calcification score did not differ significantly between groups (CaSc values 1085.2 vs 452.4 AgU; NS). The patients undergoing peritoneal dialysis showed significantly higher levels of parathyroid hormone (658.2 vs. 74.3 pg/ml; p=0.001), fibrinogen (5.82 vs. 3.89 g/l; p<0.0001) and alkaline phosphatase (330.9 vs. 168.0 U/l; p=0.001). Despite more advanced abnormalities in calcium-phosphate balance parameters and more active inflammation in peritoneal dialysis subjects we failed to demonstrate any statistically significant difference in coronary artery calcification score between patients with diabetic nephropathy on peritoneal dialysis and those with chronic kidney disease stages 2-4.

The relationships between activation of non-specific inflammatory process and malnutrition in patients on peritoneal dialysis.

Malnutrition is a frequent complication among patients on chronic peritoneal dialysis and early recognition of malnutrition can be a key factor in successful treatment. The aim of the study was to assess the nutritional status of patients on peritoneal dialysis and to search for the relationships between activation of non-specific inflammatory process and progression of malnutrition. The study group included 60 patients (age 50.4+/-14 years) on peritoneal dialysis for 17.6+/-20 months. Fourty-six patients completed the entire 24-month observation period. Nutritional status was assessed using SGA scale, anthropometric measures, bioimpendance, and several biochemical parameters. Inflammatory markers included: IL-6, TNFalpha, fibrinogen and CRP. Severe malnutrition was observed in the range between 8.4% (5 subjects, group C in SGA scale) to 11.7% (7 subjects, BMI <20 kg/m2) of patients. The nutritional status of the entire cohort was constant over 2 years of observation (based on SGA scale), although the mean albumin level decreased significantly after 24 months of observation (from 39.55+/-3.5 to 37.63+/-3.7 g/l; p<0.01). The mean concentrations of CRP (4.8 and 5.25 mg/l), IL-6 (3.45 and 6.8 pg/ml) and leptin (22.95 and 22.2 ng/ml) were above reference ranges both at the initial and final assessment. Moreover, the concentration of IL-6 significantly increased (p<0.001) after 24 months of observation. Patients treated with PD are frequently affected by malnutrition. Our results indicate a strong association between nutritional indices and markers of inflammation.

[Arterial wave velocity and coronary risk factors in patients with type-2 diabetes]. / Predkosc fali tetna a wybrane czynniki ryzyka choroby niedokrwiennej serca u pacjentów z cukrzyca typu 2.

Arterial compliance is a significant prognostic factor of cardiovascular risk. Elevated serum homocysteine level is related to increased oxidative stress and impaired endothelial function. The aim of the study was to evaluate factors influencing arterial compliance in patients with diabetes. We examined 30 patients with type 2 diabetes. In each patient we performed measurements of arterial compliance, blood pressure, serum lipids, glucose and homocysteine concentration and microalbuminuria. Serum lipids were determined by enzymatic methods, using Boehringer-Mannheim reagents, homocysteine levels by ELISA, using Bio-Rad reagents, HbA1c by HPLC method and microalbuminuria by immunonephelometric method. Arterial compliance assessment was based on measurements of pulse wave velocity (PWV), registered on femoral and carotid arteries (mean of 20 measurements), using automatic sensor (Fukuda) and Complior system. The mean age of patients was 62.2 +/- 8.1 years, BMI 30.0 +/- 3.4 kg/m2, waist to hip ratio (WHR) 0.9 +/- 0.09, mean diabetes duration 14.1 +/- 7.5 years, and HbA1c level 7.7 +/- 1.5%. Mean cholesterol, HDL-cholesterol and triglyceride serum concentrations were as follows: 5.7 +/- 1, 2 mmol/l; 1.32 +/- 0.43 mmol/ll and 1.8 +/- 1.0 mmol/l, and homocysteine levels 16.3 +/- 3.9 mmol/l. Arterial compliance significantly correlated with age (r = 0.52, p = 0.007), homocysteine concentration (r = 0.47, p = 0.015), WHR(r = 0.47, p = 0.015) and creatinine levels (r = 0.51, p = 0.016). The results of the study suggest that in patients with type 2 diabetes arterial compliance is associated not only with age but also with serum homocysteine levels and renal function parameters.