Electrochemical Sensors and Biosensors for Identification of Viruses: A Critical Review.

Due to their life cycle, viruses can disrupt the metabolism of their hosts, causing diseases. If we want to disrupt their life cycle, it is necessary to identify their presence. For this purpose, it is possible to use several molecular-biological and bioanalytical methods. The reference selection was performed based on electronic databases (2020-2023). This review focused on electrochemical methods with high sensitivity and selectivity (53% voltammetry/amperometry, 33% impedance, and 12% other methods) which showed their great potential for detecting various viruses. Moreover, the aforementioned electrochemical methods have considerable potential to be applicable for care-point use as they are portable due to their miniaturizability and fast speed analysis (minutes to hours), and are relatively easy to interpret. A total of 2011 articles were found, of which 86 original papers were subsequently evaluated (the majority of which are focused on human pathogens, whereas articles dealing with plant pathogens are in the minority). Thirty-two species of viruses were included in the evaluation. It was found that most of the examined research studies (77%) used nanotechnological modifications. Other ones performed immunological (52%) or genetic analyses (43%) for virus detection. 5% of the reports used peptides to increase the method's sensitivity. When evaluable, 65% of the research studies had LOD values in the order of ng or nM. The vast majority (79%) of the studies represent proof of concept and possibilities with low application potential and a high need of further research experimental work.

Eicosanoids in human heart failure: pilot study of plasma epoxyeicosatrienoic and dihydroxyeicosatrienoic acid levels.

Introduction: The role of eicosanoids, metabolites of arachidonic acid with cardio-renal activity, remains unclear in human heart failure (HF). Methods: We enrolled 50 patients with HF to measure plasma 14,15-EET and 14,15-DHET levels using commercial ELISA kits and compared them with 25 age- and sex-matched controls. Results: Both of the measured eicosanoids were significantly higher in the HF group: 14,15-EET (91.3 ±25.7 ng/ml vs. 64.95 ±35.4 ng/ml) and 14,15-DHET (10.58 ±2.06 ng/ml vs. 9.07 ±1.60 ng/ml), p for both < 0.001. Conclusions: We found that peripheral plasma eicosanoid (14,15-EET, 14,15-DHET) levels are raised in patients with HF compared to age- and sex-matched controls.

Serum Thymidine Kinase 1 - Potential Prostate Cancer Biomarker: A Clinical Study.

BACKGROUND/AIM: Serum thymidine kinase 1 (STK1) is a proliferation biomarker that has been used as a diagnostic marker of several malignant diseases. However, there are limited data for prostate cancer (PCa). PATIENTS AND METHODS: In this study, we retrospectively analysed serum samples from 169 patients with biopsy confirmed PCa, who had been indicated for radical prostatectomy (RP) between 2013-2016. The results were compared with those in serum samples from 39 healthy men. We used commercially available enzymatic immunoassay to determine the levels of STK1. The patients were divided into groups according to the Gleason score (GS) and risk factors for adjuvant radiotherapy (aRT), which were defined as GS 8-10, pT3, and a positive surgical margin. RESULTS: The median serum level of STK1 in PCa patients was 0.289 pmol/l. In the control group, the median value was 0.0116 pmol/l (p<0.001). By comparing the patients with GS≤6 vs. 7 vs. ≥8 (p=0.01), we found statistically significant differences. In the correlation of STK1 values with risk factors, we found statistically significant differences both in comparison of 0 vs. 1 vs. 2 vs. 3 risk factors (p=0.021), as well as ≤1 vs. 2≥ risk factors (p=0.009). CONCLUSION: The levels of STK1 are significantly higher in patients with PCa than those in healthy controls. Furthermore, STK1 values correlate with GS and predefined risk factors for aRT. Therefore, STK1 can be considered as a potential tumour marker of PCa diagnosis and risk stratification.

Impact of liver fibrosis and nodules formation on hemodynamics in young adults after total cavopulmonary connection. A magnetic resonance study.

Background: The aim of this study was to analyze the relation between the hepatic fibrosis markers, liver morphology and hemodynamics assessed by magnetic resonance imaging (MRI) after total cavopulmonary connection (TCPC). Materials and methods: Adult patients after TCPC performed in childhood between 1993 and 2003 are the subjects of this observational study. The follow-up protocol consisted of clinical and echocardiographic examination, liver elastography, cardiopulmonary exercise test, MRI hemodynamics and liver morphology assessment and direct enhanced liver fibrosis (ELF) test. Results: The cohort consisted of 39 patients (46% female) with a median age at study 26 (IQR 23-28) years and interval from TCPC 21 (IQR 20-23) years. There was no correlation between ELF test and any MRI variables, but procollagen III amino-terminal peptide (PIIINP), a single component of ELF test, correlated significantly with ventricular end-diastolic volume (r = 0.33; p = 0.042) and inferior vena cava flow (r = 0.47; p = 0.003). Fifteen (38%) patients with liver nodules had compared to other 24 patients higher end-diastolic volume (ml/m2) 102.8 ± 20.0 vs. 88.2 ± 17.7; p = 0.023, respectively. PIIINP correlated significantly with inferior vena cava flow (r = 0.56; p = 0.030) and with end-diastolic volume (r = 0.53; p = 0.043), but only in patients with liver nodules. Conclusion: Gradual progression of liver fibrosis, particularly hepatic arterialization caused by liver nodules formation, increases inferior vena cava flow and subsequent ventricular volume overload may further compromise single ventricle functional reserve in adult patients after TCPC.

Antimicrobial peptide in polymethylmethacrylate bone cement as a prophylaxis of infectious complications in orthopedics-an experiment in a murine model.

Polymethylmethacrylate (PMMA) bone cement mixed with antibiotics is used in orthopedic surgery to cope with implant-related infections which are typically associated with the formation of bacterial biofilms. Taking into account the growing bacterial resistance to current antibiotics, we examined here the efficacy of a selected antimicrobial peptide (AMP) mixed into the bone cement to inhibit bacterial adhesion and the consequent biofilm formation on its surface. In particular, we followed the formation of bacterial biofilms of methicillin-resistant Staphylococcus aureus (MRSA) on implants made from PMMA bone cement loaded with AMP composed of 12 amino acid residues. This was evaluated by CFU counting of bacteria released by sonication from the biofilms formed on their surfaces after these implants were retrieved from the infected murine femoral canals. The AMP loaded in these model implants prevented adhesion of MRSA and the subsequent formation of MRSA biofilm on the surfaces of more than 80% of these implants, whereas biofilms did form on control implants made from the plain cement. The results of our experiments performed in the murine femoral canal indicate the potential for this murine osteomyelitis model to mimic actual operations in orthopedics.

Perioperative changes of FGF23 in patients undergoing surgery for primary hyperparathyroidism.

Background: Fibroblast growth factor 23 (FGF23) is a key regulator of urine phosphate excretion. The aim of the study was to investigate the perioperative (intraoperative and postoperative) changes of plasma intact and C-terminal FGF23 (iFGF23, cFGF23) concentrations in patients with primary hyperparathyroidism (pHPT) submitted to surgery. Materials and methods: The study involved 38 adult patients with pHPT caused by adenoma. Parathyroid hormone (PTH) levels were investigated intraoperatively (just before the incision and 10 min after adenoma excision). cFGF23, iFGF23, phosphate, estimated glomerular filtration rate (eGFR), and procollagen type 1 N-terminal propetide (P1NP) were measured intraoperatively and postoperatively (next day after the surgery). Results: PTH levels decreased intraoperatively (13.10 pmol/L vs 4.17 pmol/L, P< 0.0001). FGF23 levels measured intraoperatively were at the upper level of reference interval. cFGF23 decreased postoperatively compared with the values measured just before the incision (cFGF23: 89.17 RU/mL vs 22.23 RU/mL, P< 0.0001). iFGF23 decreased as well, but the postoperative values were low. Postoperative inorganic phosphate values increased (1.03 mmol/L vs 0.8 mmol/L, P= 0.0025). We proved significant negative correlation of perioperative FGF23 with inorganic phosphate (cFGF23: Spearman's r = -0.253, P= 0.0065; iFGF23: Spearman's r = -0.245, P= 0.0085). We also found that FGF23 values just before incision correlated with eGFR (cystatin C) (cFGF23: Spearman's r = -0.499, P= 0.0014; iFGF23: Spearman's r = -0.413, P= 0.01). Conclusion: Intraoperative iFGF23 and cFGF23 did not change despite PTH decreased significantly. cFGF23 and iFGF23 significantly decreased 1 day after parathyroidectomy and are associated with increase of inorganic phosphate in pHPT patients. cFGF23 and iFGF23 just before incision correlated with eGFR (cystatin C). Similar results found in both iFGF23 and cFGF23 suggest that each could substitute the other.

Hyaluronic acid concentration in the middle ear fluid as an indicator of severity of the secretory otitis in newborns with cleft palate.

The aim of the study was to investigate the hyaluronic acid concentration in middle ear fluid of patients with cleft palate as an indicator of the severity of the disease. Hyaluronic acid was examined in the middle ear fluid of 65 children (48 boys and 17 girls) subjected to cleft lip surgery in neonatal period up to 10 days of age. Patients were divided into 3 groups according to the course of the disease. First group consists of 15 patients with favorable course, second group consist of 25 patients with moderate course, third group included 25 patients with an adverse course. Hyaluronic acid levels were determined by commercially available immunoassay. The concentrations of hyaluronic acid in the middle ear fluid were as follows (mean+SEM): favorable course: 14253+2393 µg/l, moderate course: 7503+1345 µg/l, adverse course: 5905+2393 µg/l. Patients with adverse course and moderate course had significantly decreased hyaluronic acid levels in middle ear fluid compared to the patients with favorable course (P=0.02 and P=0.0018). Hyaluronic acid concentration is related to the course of the disease and the lowest values are most frequent in patients with an adverse course.

Thymidine Kinase-1 as Additional Diagnostic Marker of Prostate Cancer.

BACKGROUND: Thymidine kinase-1 (TK-1) is associated with proliferation and malignancy and has been extensively studied as a diagnostic biomarker for a variety of tumors, but there are limited data for prostate cancer. METHODS: TK-1 concentrations in serum were measured in 59 patients with prostate cancer (mean age 68 years) and in the control group of 28 healthy men (mean age 63 years) using commercially available enzymatic immunoassay (LSBio, Inc., Seattle, WA, USA). The patients were divided with respect to the severity of the disease into two groups according to the European Association of Urology (EAU) guidelines (Stage 1, 2 - less severe tumors, stage 3 - severe tumors). RESULTS: Serum thymidine kinase-1 concentrations were significantly elevated in the group of the patients with prostate cancer compared to the healthy individuals (0.204 pmol/L vs. 0.072 pmol/L, with p < 0.0001). Diagnostic efficiency of serum TK-1 concentrations was 0.792 with the specificity of 53.6% and sensitivity of 94.9%. Patients with less severe tumors (Stage 1, 2) and severe tumors (Stage 3) had significantly increased levels of TK-1 as well (p < 0.0001). Combination of TK-1 and PSA investigation in patients with PCa improve the diagnostic validity of TK-1 (AUC = 0.87). CONCLUSIONS: Concentrations of thymidine kinase 1 are increased in all patients with prostate cancer and even more in patients with severe prostate cancer. Thymidine kinase 1 appears to be a promising additional diagnostic marker promising in patients with prostate cancer.

Diagnostic benefits of mindin as a prostate cancer biomarker: Dijagnosticke prednosti mindina kao biomarkera raka prostate.

BACKGROUND: It has been shown that decreased expression and activity of extracellular matrix protein mindin correlate with various types of cancers including breast, colon and lung cancers. The aim of the presented study was to investigate the serum mindin levels in prostate cancer. METHODS: Mindin concentrations in serum were measured in 56 patients with prostate cancer (mean age 68 years) and in control group of 29 healthy men (mean age 64 years) using commercially available enzymatic immunoassay (Cusabio, WuHan, China). The patients were divided with respect to the severity of the disease into two groups according to the EAU guidelines (stage 1, 2 - less severe tumours, stage 3, 4 - severe tumours). RESULTS: Serum mindin concentrations were significantly elevated in the group of healthy individuals unlike in the patients with prostate cancer (2.12 ng/mL vs 0.78 ng/mL, with P=0.0007, AUC=0.705). Patients with less severe tumours (stage 1, 2) and severe tumours (stage 3, 4) had significantly decreased levels of S-mindin as well (P=0.0037), although the difference in serum mindin concentrations between the patients with less severe and severe tumours was not significant. CONCLUSIONS: Concentrations of mindin were decreased in patients with prostate cancer and reduced in patients with less severe prostate cancer as well. Mindin appears to be a promising diagnostic marker useful in the diagnosis of prostate cancer.

Polycyanoacrylate (super glue) as bladder concretion in a patient after bilateral inguinal hernioplasty.

A rare case of cyanoacrylate urine bladder urolithiasis in a 60-year-old male is presented. The application of surgical glue (Glubran) as treatment of seroma one month after laparoscopic inguinal hernioplasty led to the instillation of the n-butyl cyanoacrylate into the bladder resulting in the formation of a concretion. Infrared spectroscopy of the urine stone removed by cystoscopic laser lithotripsy four months after the surgery allowed the identification of the nature of the stone and revealed cyanoacrylate as the major component and co-monomer methacryloxy sulfolane as the minor component. Polypropylene from the mesh was not detected.

High-Performance Liquid Chromatography as a Novel Method for the Determination of α-Defensins in Synovial Fluid for Diagnosis of Orthopedic Infections.

The α-defensins (AD) present in synovial fluid have been regarded as constituting the most accurate periprosthetic joint infection (PJI) biomarker. The methods most commonly used for estimating AD as a biomarker are the qualitative Synovasure® PJI tests, based on the technique of lateral flow, and quantitative enzyme-linked immunosorbent assay (ELISA). Here, we propose a novel test based on detecting α-defensins in synovial fluid by high-performance liquid chromatography (HPLC). Synovial fluid was collected from 157 patients diagnosed with PJI, infectious arthritis (IA), arthrosis, reactive arthritis, and rheumatoid arthritis. AD concentrations in the fluid were determined by HPLC, and these same samples were used for additional diagnostic analyses. The results were statistically processed to calculate cutoff concentrations for PJI and IA. HPLC testing showed a sensitivity of 94% and a specificity of 92% for diagnosis of PJI, and a sensitivity of 97% and a specificity of 87% for diagnosis of IA. Using HPLC, we detected in synovial fluid a combination of three α-defensins: human neutrophil peptides HNP1, HNP2, and HNP3. All measured AD concentration values shown in this work refer to the sum of these three individual concentrations. Our study shows that the HPLC method meets the conditions for measuring precise concentrations of the sum of AD and can be recommended as part of a diagnostic array for PJI and IA diagnostics. By this method, we have verified that higher levels of AD in synovial fluid can also be seen in rheumatoid illnesses, crystal arthropathies, and reactive arthritis.

The Effect of Inhibin A on Prenatal Screening Results for Down Syndrome in the High Risk Czech Pregnant Women.

BACKGROUND: Currently, prenatal testing is based on an ultrasound examination, the testing of certain biochemical markers and, most recently, also on the analysis of fragments from the extracellular DNA of the fetus in the mother´s blood. The aim of this work was to verify whether inhibin A testing during pregnancy can help influence the risk distribution of Down syndrome screening results in high risk population and thus possibly reduce the number of unnecessarily invasive procedures, or for better stratification of risks when deciding on non-invasive DNA testing. METHODS: The concentrations of inhibin A were measured using a chemiluminescent immunoassay in two groups of screening tests. The first group (triple test) included a total of 277 pregnant women; the second group (integrated test) included 91 pregnant women. Risk assessments of screenings were performed using Alpha software, LMS. RESULTS: The resulting risk for pregnant women without the determination of inhibin A was higher or equal to 1:300 (triple test) and 1:150 (integrated test). Inhibin A was then measured in the monitored groups and the risk was recalculated. In the first group (triple test) the risk was lower than 1:300 in 152 pregnant women and in the other group (the integrated test) in 47 pregnant women. At the end of the study, all results were compared with the outcome of the pregnancy. CONCLUSIONS: The results obtained show that the inclusion of inhibin A in screening protocols reduces the number of positive results in high risk population screened without inhibin A.

Manganese and selenium concentrations in cerebrospinal fluid of seriously ill children.

BACKGROUND: The homeostasis of essential trace elements such as selenium and manganese may be altered in patients with severe diseases of various etiologies (trauma brain injuries, tumors, leukemias, lymphomas, neurological diseases). METHODS: Concentration of manganese and selenium were determined in cerebrospinal fluid by electrothermal atomic absorption spectrometry in 50 hospitalized children with various clinical ethiologies including oncological, neurological, and brain related diseases. RESULTS: The concentrations of manganese in cerebrospinal fluid of children were 0.97±0.67 µg/L. The concentrations of selenium were 13.3±3.5 µg/L. The concentrations were similar as published in adults. The values did not correlated with the age, gender and severity of the disease. CONCLUSION: We evaluated values of selenium and manganese in cerebrospinal fluid of seriously diseased children.

A Comparison of Four Methods (Immunochemistry and HPLC) for Determination of 25-(OH)-Vitamin D in Postmenopausal Women.

BACKGROUND: Three immunochemical methods for the determination of 25-(OH)-vitamin D and validated HPLC method for the determination of 25-(OH)-vitamin D3 and 25-(OH)-vitamin D2 were compared. 62 patient samples from postmenopausal women were measured and the results obtained by all these methods were compared. METHODS: We used three chemiluminescent assays for determination of 25-(OH)-vitamin D. 25-(OH)-vitamin D3 and 25-(OH)-vitamin D2 were determined by HPLC with UV detection (Agilent 1200). The chemiluminescent assays were performed using the Abbott Architect i4000SR analyzer (Abbott Laboratories, Germany), the ADVIA Centaur (Siemens, USA), and the Liaison XL (DiaSorin Inc, USA). The statistical evaluation was done using GraphPad Prism 6.0. RESULTS: The data were tested by Tukey's multiple comparison test. All methods showed significant differences in comparison with the immunochemical method from DiaSorin (p < 0.001 for Abbott, p < 0.05 for Siemens, and p < 0.0001 for HPLC). The comparison of the immunochemical method from Siemens with HPLC was also significant, p < 0.05. The mean of DiaSorin measurements was 38% lower than the mean of HPLC measurements. The non-significant difference was shown by the comparison of Abbott with HPLC and also Abbott with Siemens. Means for the 25-(OH)-vitamin D methods used were: Abbott 70.2 ± 24.2 nmol/L, Siemens 67.6 ± 27.9 nmol/L, DiaSorin 53.5 ± 17.1, and HPLC 82.4 ± 40.0 nmol/L. CONCLUSIONS: The comparison of the DiaSorin immunochemical assay with other tested methods showed the greatest deviation. The mean of DiaSorin measurements was 38% lower than the mean of HPLC measurements. According to the results of the DiaSorin method, most patients treated with vitamin D would not achieve the optimal level of 25-(OH)-vitamin D and this could negatively affect the clinical decision.

Determination of Urine Albumin by New Simple High-Performance Liquid Chromatography Method.

BACKGROUND: A simple high-performance liquid chromatography (HPLC) method was developed for the determination of albumin in patients' urine samples without coeluting proteins and was compared with the immunoturbidimetric determination of albumin. Urine albumin is important biomarker in diabetic patients, but part of it is immuno-nonreactive. METHODS: Albumin was determined by high-performance liquid chromatography (HPLC), UV detection at 280 nm, Zorbax 300SB-C3 column. Immunoturbidimetric analysis was performed using commercial kit on automatic biochemistry analyzer COBAS INTEGRA® 400, Roche Diagnostics GmbH, Manheim, Germany. RESULTS: The HLPC method was fully validated. No significant interference with other proteins (transferrin, α-1-acid glycoprotein, α-1-antichymotrypsin, antitrypsin, hemopexin) was found. The results from 301 urine samples were compared with immunochemical determination. We found a statistically significant difference between these methods (P = 0.0001, Mann-Whitney test). CONCLUSION: New simple HPLC method was developed for the determination of urine albumin without coeluting proteins. Our data indicate that the HPLC method is highly specific and more sensitive than immunoturbidimetry.

Diagnostic Efficiency of Serum and Urine Procathepsin B and Cathepsin B in Patients with Carcinoma of the Urinary Bladder.

BACKGROUND: The aim of the study was to evaluate the diagnostic efficiency of cathepsins B (cathepsin B and procathepsin B) in patients with transient cell carcinoma of the urinary bladder. METHODS: Serum and urine concentrations of cathepsin B and procathepsin B were measured by two commercially available enzymatic immunoassays in a group of 125 patients with bladder cell carcinoma without metastases and in a group of 72 healthy individuals. Concentrations in urine were adjusted to creatinine. RESULTS: Concentrations of both cathepsin B and procathepsin B in serum and urine were significantly elevated in patients with bladder cell carcinoma (p < 0.0001 for U-procathepsin B, U-procathepsin B/creatinine, and U-cathepsin B/creatinine, p = 0.0001 for U-cathepsin B, p = 0.0002 for S-procathepsin B and p = 0.02 for S-cathepsin B). Comparison of all diagnostic efficiencies of cathepsin B and procathepsin B in serum and in urine showed the best diagnostic accuracy for procathepsin B in urine (AUC = 0.81 vs. 0.50). The ratio of U-procathepsin B/creatinine was also more efficient than the ratio of U-cathepsin B/creatinine (AUC = 0.81 vs. AUC = 0.70). The diagnostic efficiencies of both parameters in serum were low (S-procathepsin B: AUC = 0.50, S-cathepsin B: AUC = 0.60). U-procathepsin B and U-procathepsin B/creatinine ratio show significantly better diagnostic efficiency in patients with invasive bladder tumors than other parameters (S-procathepsin B, S-cathepsin B, U-cathepsin B and U-Cathepsin B/creatinine; U-procathepsin B: AUC = 0.82, U-procathepsin B/creatinine: AUC = 0.86, S-procathepsin B and cathepsin B: AUC = 0.51 - 0.68). CONCLUSIONS: Procathepsin B concentration in urine is a valuable diagnostic marker in patients with bladder cell carcinoma.

Usefulness of N-Terminal Pro-Brain Natriuretic Peptide to Predict Mortality in Adults With Congenital Heart Disease.

Natriuretic peptides are often elevated in congenital heart disease (CHD); however, the clinical impact on mortality is unclear. The aim of our study was to evaluate the prognostic value of N-terminal pro-brain natriuretic peptide (NT-proBNP) in the prediction of all-cause mortality in adults with different CHD. In this prospective longitudinal mortality study, we evaluated NT-proBNP in 1,242 blood samples from 646 outpatient adults with stable CHD (mean age 35 ± 12 years; 345 women). Patients were followed up for 6 ± 3 (1 to 10) years. The mortality rate was 5% (35 patients, mean age 40 ± 14 years, 17 women). Median NT-proBNP (pg/ml) was 220 in the whole cohort, 203 in survivors, and 1,548 in deceased patients. The best discrimination value for mortality prediction was 630 pg/ml with 74% sensitivity and 84% specificity. During the follow-up, the survival rate was 65% for those with median NT-proBNP ≥630 pg/ml and 94% for NT-proBNP <630 pg/ml; p <0.0001. There was only 1% mortality among 388 patients with at least 1 NT-proBNP value ≤220 pg/ml compared with 41% mortality among 54 patients with at least 1 NT-proBNP value >1,548 pg/ml. Even the first (baseline) measurements of NT-proBNP were strongly associated with a high risk of death (log10 NT-proBNP had hazard ratio 7, p <0.0001). In conclusion, NT-proBNP assessment is a useful and simple tool for the prediction of mortality in long-term follow-up of adults with CHD.

NT-proBNP levels and their relationship with systemic ventricular impairment in adult patients with transposition of the great arteries long after Mustard or Senning procedure.

BACKGROUND: The aim of the study was to investigate serum NT-proBNP levels in adult patients with transposition of the great arteries (d-TGA) corrected by atrial switch procedures (Mustard or Senning) operation and to assess the relationship with ventricular impairment and NYHA class. METHODS: Serum NT-proBNP levels were measured in a group of 81 consecutive adult patients (59 males, mean age 27 years and 22 females, mean age 28 years) with transposition of the great arteries (TGA) after surgical correction in childhood, and in a control group of 25 healthy individuals (16 males, mean age 32 years, and 9 females, mean age 29 years). Age-matched correlation of NT-proBNP concentrations in TGA patients after Mustard or Senning correction was performed, but this correlation was considered not significant (p=0.08). RESULTS: Concentrations of NT-proBNP in patients with TGA were significantly elevated compared to the control group of healthy individuals (203 ng/L vs. 41 ng/L, p<0.0001). Patients after the Mustard repair had significantly higher NT-proBNP values than patients after the Senning operation (234 ng/L vs. 148 ng/L, p=0.0023). NT-proBNP correlated negatively with the systemic right ventricular ejection fraction with the greatest significance in patients after Mustard correction (r=-0.32, p<0.0001). The concentration of NT-proBNP was also associated with NYHA functional class (p=0.0035) with the greatest significance in patients with Mustard correction (p=0.028). CONCLUSIONS: Elevated levels of NT-proBNP appear to be a useful tool in assessing heart failure in patients with transposition of the great arteries after atrial switch correction.

Concentrations of MMP-9 and TIMP-1 in lip tissue and their impact on cleft lip surgery healing.

AIM: To compare aspects of wound healing after cleft lip surgery performed within one week of age and wound healing after surgery performed within 2 - 4 months of age, especially concentrations of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinases-1 (TIMP-1) in tissue removed during surgery. METHODS: 34 tissue samples (26 boys and 8 girls) were removed during surgery within one week of age (n=19) or within 2 - 4 months of age (n=15). Tissue samples were separated into epidermis, dermis and mucous membrane. Proteins were extracted in cacodylic buffer for 24 h at a temperature 2 - 8 ºC. Total protein concentrations were examined using a modification of the Lowry method. Samples were examined using ELISA kit Amersham Biotrak Activity Assay (GE Healthcare UK) for detection of MMP-9 and TIMP-1 concentrations. RESULTS: MMP-9: early surgery - epidermis 2.168 ± 3.303 µg/g of protein (mean ± SD), dermis 1.251 ± 1.848 µg/g, 2 - 4 months surgery - epidermis 0.347 ± 0.212 µg/g, dermis 0.555 ± 0.276 µg/g. TIMP-1: early surgery - epidermis 1.762 ± 2.162 µg/g, dermis 1.628 ± 0.822 µg/g, mucous membrane 2.066 ± 1.717 µg/g, 2 - 4 months surgery - epidermis 1.881 ± 2.810 µg/g, dermis 3.117 ± 1.540 µg/g, mucous membrane 4.833 ± 6.550 µg/g. CONCLUSIONS: There were no significant differences in concentrations of protein MMP-9 in epidermis and dermis and TIMP-1 in epidermis and mucous membrane according to time of surgery. Significantly decreased levels of TIMP-1 in dermis were found in samples obtained from early surgery compared to levels in samples obtained from 2 - 4 months surgery.