BACKGROUND Acute kidney injury (AKI) is a common cause of organ failure in patients after major trauma and is associated with increased morbidity and mortality. Early identification of patients at risk enables the implementation of a bundle of supportive care, which reduces the incidence of AKI. The primary objective of our study was to investigate whether the levels of biomarkers on admission predicted the onset of early AKI in patients with serious injuries. MATERIAL AND METHODS This prospective observational study included 98 adult patients of both sexes with a serious injury (injury severity score >16). At admission, blood samples were taken, and creatinine, neutrophil gelatinase-associated lipocalin (NGAL), high mobility group box 1 (HMGB-1), and markers of rhabdomyolysis (creatine kinase, myoglobin) were evaluated. The patients were provided with standard resuscitation care, and the occurrence of AKI was monitored during the first 7 days after admission to the Intensive Care Unit, according to the Kidney Disease Improving Global Outcomes diagnostic criteria. RESULTS AKI occurred in 25 (25.5%) patients, in whom the admission levels of HMGB-1, NGAL, creatinine, and myoglobin were significantly higher than in non-AKI patients (48.3±98.4 vs 113.0±209.4 µg/L, P=0.006; 150.2±349.9 vs 181.4±152.2 µg/L, P=0.004; 83.1±20.8 vs 118.8±32.2 µmol/L, P<0.005; 2734.4±2214.5 vs 4182.3±2477.1 µg/L, P=0.008, respectively). Creatine kinase was 14.5±9.2 µkat/L in non-AKI patients and 13.7±7.9 µkat/L in AKI patients (P=0.916). CONCLUSIONS Admission levels of HMGB-1, NGAL, creatinine, and myoglobin predicted the risk of AKI in severely injured patients.

• MeSH
• akutní poškození ledvin * diagnóza   etiologie   MeSH
• biologické markery MeSH
• dospělí MeSH
• kreatinin MeSH
• kreatinkinasa MeSH
• lidé MeSH
• lipokalin-2 MeSH
• myoglobin * MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• pozorovací studie MeSH
• Názvy látek
• biologické markery MeSH
• kreatinin MeSH
• kreatinkinasa MeSH
• lipokalin-2 MeSH
• myoglobin * MeSH

BACKGROUND: Through regulation of signaling pathways, microRNAs (miRNAs) can be involved in sepsis and associated organ dysfunction. The aims of this study were to track the 7-day time course of blood miRNAs in patients with sepsis treated with vancomycin, gentamicin, or a non-nephrotoxic antibiotic and miRNA associations with neutrophil gelatinase-associated lipokalin (NGAL), creatinine, procalcitonin, interleukin-6, and acute kidney injury (AKI) stage. METHODS: Of 46 adult patients, 7 were on vancomycin, 20 on gentamicin, and 19 on another antibiotic. Blood samples were collected on days 1, 4, and 7 of treatment, and miRNAs were identified using quantitative reverse transcription PCR. RESULTS: The results showed no relationship between miRNA levels and biochemical variables on day 1. By day 7 of gentamicin treatment miR-15a-5p provided good discrimination between AKI and non-AKI (area under curve, 0.828). In patients taking vancomycin, miR-155-5p and miR-192-5p positively correlated with creatinine and NGAL values, and miR-192-5p and miR-423-5p positively correlated with procalcitonin and interleukin-6 in patients treated with a non-nephrotoxic antibiotic. In patients together we found positive correlation between miR-155-5p and miR-423-5p and all biochemical markers. CONCLUSION: The results suggest that these four miRNAs may serve as diagnostic or therapeutic tool in sepsis, renal injury and nephrotoxic treatment. TRIAL REGISTRATION: ClinicalTrials.gov , ID: NCT04991376 . Registered on 27 July 2021.

• Klíčová slova
• Acute kidney injury, Gentamicin, Nephrotoxicity, Sepsis, Vancomycin, microRNA,
• MeSH
• akutní poškození ledvin * komplikace   MeSH
• antibakteriální látky terapeutické užití   MeSH
• cirkulující mikroRNA * MeSH
• dospělí MeSH
• gentamiciny MeSH
• interleukin-6 metabolismus   MeSH
• kreatinin MeSH
• lidé MeSH
• lipokalin-2 MeSH
• mikro RNA * genetika   MeSH
• prokalcitonin MeSH
• sepse * komplikace   MeSH
• vankomycin terapeutické užití   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• klinická studie MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antibakteriální látky MeSH
• cirkulující mikroRNA * MeSH
• gentamiciny MeSH
• interleukin-6 MeSH
• kreatinin MeSH
• lipokalin-2 MeSH
• mikro RNA * MeSH
• prokalcitonin MeSH
• vankomycin MeSH

PURPOSE OF THE STUDY Laboratory methods are central to prosthetic joint infection (PJI) diagnosis. Most research teams focus on detection of specific inflammatory markers, causative pathogens, or on assessment of the tissue response. This study sought to determine the optimal cut-off values and diagnostic performance of selected synovial markers in relation to the diagnosis of hip or knee PJI. The studied markers were synovial level of glucose, lactate, coefficient of energy balance (CEB) and NGAL (neutrophil gelatinase-associated lipocalin). MATERIAL AND METHODS This prospective study includes 89 patients who underwent revision total knee or hip arthroplasty for septic or aseptic reasons in the period from 2014 to 2017. Among these 89 patients, there are 2 cases of prosthetic hip infection, 22 cases of prosthetic knee infection, 31 aseptic revision total hip arthroplasties and 34 aseptic revision total knee arthroplasties. The diagnostic characteristics of the studied methods were set in relation to the reference standard, the 2013 MSIS (Musculoskeletal Infection Society) criteria. The cut-off values were calculated using the ROC (receiver operating characteristic curve) analysis. RESULTS The synovial glucose test is considered positive if the glucose level drops below 2.65 mmol/L. The area under the curve is 0.813, sensitivity 75.0%, specificity 83.1%. The synovial lactate test is considered positive if lactate level rises above 8.87 mmol/L. The area under the curve is 0.882, sensitivity 70.8%, specificity 95.4%. Synovial NGAL is considered positive if its level exceeds 998 μg/L. The area under the curve is 1.000, sensitivity 100.0%, specificity 100.0%. CEB is considered positive if its value is lower than +4.665. The area under the curve is 0.883, sensitivity 91.7% and specificity 69.8%. Combining of these tests with other synovial markers does not improve the diagnostic performance of the studied tests. CONCLUSIONS The glucose and lactate levels and CEB undoubtedly reflect the presence of an inflammatory process in a prosthetic joint. However, the diagnostic characteristics of these tests are not better than those of other modern diagnostic techniques. As opposed to these tests, synovial NGAL shows excellent diagnostic performance. Nonetheless, the potential of this method shall be verified on larger cohorts of patients. Key words: prosthetic joint infection, periprosthetic infection, total knee arthroplasty, total hip arthroplasty, diagnosis, glucose, lactate, CEB, NGAL.

• MeSH
• biologické markery analýza   MeSH
• C-reaktivní protein analýza   MeSH
• glukosa MeSH
• infekce spojené s protézou * diagnóza   etiologie   MeSH
• kyselina mléčná MeSH
• lidé MeSH
• lipokalin-2 analýza   MeSH
• náhrada kyčelního kloubu * škodlivé účinky   MeSH
• prospektivní studie MeSH
• protézy kolene * škodlivé účinky   MeSH
• senzitivita a specificita MeSH
• synoviální tekutina chemie   MeSH
• totální endoprotéza kolene * škodlivé účinky   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• biologické markery MeSH
• C-reaktivní protein MeSH
• glukosa MeSH
• kyselina mléčná MeSH
• lipokalin-2 MeSH

Cisplatin is a commonly used chemotherapeutic drugs. It is known for its nephrotoxic side effects with an increased risk of acute kidney injury. Finding of clinically feasible cisplatin nephrotoxicity markers is of importance. In our study, we compared neutrophil gelatinase-associated lipocalin (NGAL) in serum and urine, the estimated glomerular filtration rate (based on serum cystatin C) and urine albumin as markers of nephrotoxicity. The study involved 11 men and 9 women (mean ± SD age 58.2±9.5 years) with different malignancies treated with cisplatin in four cycles of chemotherapy (I - IV). Samples 0-4 were taken before, immediately after, in 3, 6 and 24 hours after administering chemotherapy. We detected significant increase of ACR in Sample 2 (p=0.03) and decrease of eGFR in Sample 4 (p=0.03) up to 24 hours after cisplatin administration in the first chemotherapy cycle only. When cumulative effect of cisplatin was assessed, significantly increased values of urine albumin (vs cycle I) were found in Sample 0 (p=0.00058), 1 (p=0.00256), 2 (p=0.00456), 3 (p=0.00006) and 4 (p=0.00319) in cycles II to IV. We found a correlation between values of urine NGAL and urine albumin (r=0.68, p<0.0001). In conclusion, urine albumin was the only measured marker that consistently and statistically significantly increased after cisplatin containing chemotherapy cycles.

• MeSH
• antitumorózní látky terapeutické užití   MeSH
• cisplatina terapeutické užití   MeSH
• cystatin C moč   MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• lidský sérový albumin moč   MeSH
• lipokalin-2 moč   MeSH
• nádorové biomarkery moč   MeSH
• nádory farmakoterapie   moč   MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antitumorózní látky MeSH
• cisplatina MeSH
• CST3 protein, human MeSH Prohlížeč
• cystatin C MeSH
• LCN2 protein, human MeSH Prohlížeč
• lidský sérový albumin MeSH
• lipokalin-2 MeSH
• nádorové biomarkery MeSH

BACKGROUND AND OBJECTIVES: Renal elimination of amikacin and other aminoglycosides is slowed down in sepsis-induced acute kidney injury increasing the risk of adverse effects. Since neutrophil gelatinase-associated lipocalin (NGAL) and aminoglycosides share the mechanisms for renal excretion, the predictive power of NGAL was examined towards the changes in amikacin pharmacokinetics during early endotoxemia in anesthetized Wistar rats. METHODS: Endogenous biomarkers of inflammation and acute kidney injury were assessed including NGAL in saline-injected controls and two groups of rats challenged with an intravenous injection of bacterial lipopolysaccharide (5 mg/kg)-a fluid-resuscitated group (LPS) and a fluid-resuscitated group infused intravenously with 8 μg/kg/h terlipressin (LPS-T). Sinistrin and amikacin were infused to measure glomerular filtration rate (GFR) and amikacin clearance (CLam). The investigations included blood gas analysis, chemistry and hematology tests and assessment of urine output, creatinine clearance (CLcr) and sinistrin clearance (CLsini). RESULTS: Within 3 h of injection, systemic and renal inflammatory responses were induced by lipopolysaccharide. Gene and protein expression of NGAL was increased in the kidneys and the concentrations of NGAL in the plasma (pNGAL) and urine rose 4- to 38-fold (P < 0.01). The decreases in CLam and the GFR markers (CLcr, CLsini) were proportional, reflecting the extent to which endotoxemia impaired the major elimination mechanism for the drug. Terlipressin attenuated lipopolysaccharide-induced renal dysfunction (urine output, CLcr, CLsini) and accelerated CLam. The pNGAL showed a strong association with the CLsini (rs = - 0.77, P < 0.0005). Concerning prediction of CLam, pNGAL was comparable to CLcr (mean error - 24%) and inferior to CLsini (mean error - 6.4%), while the measurement of NGAL in urine gave unsatisfactory results. CONCLUSIONS: During early endotoxemia in the rat, pNGAL has a moderate predictive ability towards CLam. Clinical studies should verify whether pNGAL can support individualized dosing of aminoglycosides to septic patients.

• MeSH
• akutní poškození ledvin krev   MeSH
• amikacin krev   metabolismus   farmakokinetika   MeSH
• biologické markery krev   MeSH
• cytokiny MeSH
• endotoxemie chemicky indukované   MeSH
• hodnoty glomerulární filtrace fyziologie   MeSH
• krysa rodu Rattus MeSH
• ledviny patofyziologie   MeSH
• lipokalin-2 krev   metabolismus   moč   MeSH
• lipopolysacharidy farmakologie   MeSH
• metabolická clearance MeSH
• moč MeSH
• modely u zvířat MeSH
• oligosacharidy farmakokinetika   MeSH
• potkani Wistar * MeSH
• prediktivní hodnota testů MeSH
• sepse farmakoterapie   metabolismus   MeSH
• zánět MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• amikacin MeSH
• biologické markery MeSH
• cytokiny MeSH
• lipokalin-2 MeSH
• lipopolysacharidy MeSH
• oligosacharidy MeSH
• sinistrin MeSH Prohlížeč

Common variable immunodeficiency disorders (CVID) represent a group of primary immunodeficiency diseases characterized by hypogammaglobulinemia and impaired specific Ab response, resulting in recurrent infections due to dysfunctional immune response. The specific mechanisms mediating immune deficiency in CVID remain to be determined. Previous studies indicated that immune dysregulation in CVID patients is associated with chronic microbial translocation, systemic immune activation, and altered homeostasis of lymphocytic and myeloid lineages. A detailed phenotypic, functional characterization of plasma markers and immune cell populations was performed in 46 CVID patients and 44 healthy donors. CVID patients displayed significantly elevated plasma levels of a marker of neutrophil activation neutrophil gelatinase-associated lipocalin. Neutrophils from CVID patients exhibited elevated surface levels of CD11b and PD-L1 and decreased levels of CD62L, CD16, and CD80, consistent with a phenotype of activated neutrophils with suppressive properties. Neutrophils from CVID patients actively suppressed T cell activation and release of IFN-γ via the production of reactive oxygen species. Furthermore, CVID was associated with an increased frequency of low-density neutrophils (LDNs)/granulocytic myeloid-derived suppressor cells. LDN/granulocytic myeloid-derived suppressor cell frequency in CVID patients correlated with reduced T cell responsiveness. Exogenous stimulation of whole blood with bacterial LPS emulated some but not all of the phenotypic changes observed on neutrophils from CVID patients and induced neutrophil population with LDN phenotype. The presented data demonstrate that neutrophils in the blood of CVID patients acquire an activated phenotype and exert potent T cell suppressive activity. Specific targeting of myeloid cell-derived suppressor activity represents a novel potential therapeutic strategy for CVID.

• MeSH
• aktivace neutrofilů MeSH
• antigeny CD11b metabolismus   MeSH
• antigeny CD274 metabolismus   MeSH
• běžná variabilní imunodeficience imunologie   MeSH
• dospělí MeSH
• granulocyty fyziologie   MeSH
• imunologická tolerance MeSH
• kultivované buňky MeSH
• lidé středního věku MeSH
• lidé MeSH
• lipokalin-2 krev   MeSH
• mladý dospělý MeSH
• myeloidní supresorové buňky fyziologie   MeSH
• neutrofily fyziologie   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• T-lymfocyty imunologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny CD11b MeSH
• antigeny CD274 MeSH
• CD274 protein, human MeSH Prohlížeč
• LCN2 protein, human MeSH Prohlížeč
• lipokalin-2 MeSH
• reaktivní formy kyslíku MeSH

BACKGROUND/AIMS: The rate of incidence and prevalence of acute kidney injury is increasing due to an increased number of patients with heart failure. Therefore it is very pertinent to early detect the level of renal injuries and to make necessary heart failure predictions. Thus the aim of this study is to determine renal functions and prognosis stratification in chronic heart failure patients and importance of Neutrophil Gelatinase-Associated Lipocalin (NGAL), an early diagnostic marker of acute kidney injury, as well as stratification of cardiovascular risk in heart failure patients. METHODS: Data including age, gender, comorbidities and medical history of outpatients and hospitalized patients from Farmacology and NeuroHumoraL activation (FAR NHL) multicenter prospective registry comprising three Cardiological Centers in the Czech Republic were collected between 1st October 2014 and 30th November 2015. One-year follow-up data were collected in November 2016 in such a way that all patients had at least one-year data from the time of recruitment, but up to two years to the time of follow-up. One-year data were used for the whole set of patients while data up to 24 months were used with Kaplan-Meier's survival curves to analyse the patients' survival data. Blood samples were collected from the patients and basic parameters were evaluated in order to analyse Neutrophil gelatinase-associated lipocalin (NGAL) and plasma levels of N-terminal pro-brain natriuretic peptide (NT-ProBNP) using Lipocalin-2/NGAL Human ELISA kit (Bio Vendor, Czechia) and the Cobas E411 NT-proBNP Immunoassay kit (Roche Diagnostics, Indianapolis, IN, USA) respectively. Statistical analysis was further carried out to explain the level of significance of the evaluated parameters using Spearman Correlation, Mann Whitney or Kruskal-Wallis test and log-rank test. RESULTS: Out of 547 patients from Farmacology and NeuroHumoraL activation (FAR NHL) multicenter prospective registry with available data on hospitalizations, mortality, biomarkers and one-year follow-up that were recorded, there were 439 males (80.3%) with a median age of 66 years. At least one-month stable patients with left ventricle ejection fraction (LV EF) under 50% were recorded. The etiology of heart failure was ischemic heart disease in 54%, dilated cardiomyopathy in 40% and others in 6%. 69% patients were in New York Heart Association functional class II. There were 76 events (13.9%; all-cause mortality, acute heart failure hospitalization, left ventricle assist device implantation and orthotopic heart transplant) in the first 365 days of follow-up. The area under the receiver operating characteristic curve was higher for NT-proBNP (0.77) than the creatinine (0.57), NGAL (0.55) or creatinine clearance (0.54). In multivariable analyses, NT-proBNP (P= 0.001) and NGAL (P = 0.004) were significant predictors of events. Subjects with NT-proBNP and NGAL above the cut off value (NT-proBNP 1,121 pg/ml, NGAL 80 ng/ml) survived without any event in 55.7%, subjects with NT-proBNP and NGAL under the cut off value survived without any event in 90.5%, after two years (P = 0.001). CONCLUSION: The findings of the study showed that NGAL associated with NT-proBNP was a stronger predictor of the primary endpoint than NGAL or NT-proBNP alone. The level of NGAL was rising in hypertension, ischemia, anemia, hypoalbuminemia, diabetes or arrhythmias.

• Klíčová slova
• Chronic heart failure, Creatinine, N-terminal pro-brain natriuretic peptide, Neutrophil Gelatinase-associated Lipocalin, Renal function,
• MeSH
• akutní poškození ledvin komplikace   epidemiologie   MeSH
• chronická nemoc MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• lipokalin-2 krev   MeSH
• natriuretický peptid typu B krev   MeSH
• peptidové fragmenty krev   MeSH
• prognóza MeSH
• registrace MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• srdeční selhání komplikace   diagnóza   epidemiologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• multicentrická studie MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• LCN2 protein, human MeSH Prohlížeč
• lipokalin-2 MeSH
• natriuretický peptid typu B MeSH
• peptidové fragmenty MeSH
• pro-brain natriuretic peptide (1-76) MeSH Prohlížeč

OBJECTIVE: We aimed at assessing the predictive value of plasmatic Neutrophil Gelatinase Associated Lipocalin (pNGAL) at admission and severity scores to predict major adverse kidney events (MAKE, defined as death and/or need for renal replacement therapy (RRT) and/or non-renal recovery at day 90) in critically ill burn patients. MATERIAL AND METHODS: Single-center cohort study in a burn critical care unit in a tertiary center, including all consecutive severely burn patients (total burned body surface >20%) from January 2012 until January 2015 with a pNGAL dosage at admission. Reclassification of patients was assessed by Integrated Discrimination Improvement (IDI). MEASUREMENTS AND RESULTS: 87 patients were included. Mean age was 47.7 (IQ 25-75: 33.4-65.2) years; total burn body surface area was 40 (IQ 25-75: 30-55) % and ICU mortality 36%. 39 (44.8%) patients presented a MAKE, 32 (88.9%) patients died at day 90. pNGAL was higher in the MAKE group (423 [IQ 25-75: 327-518]pg/mL vs 184 [IQ 25-75: 147-220]pg/mL, p<0.001). In multivariate analysis, pNGAL and abbreviated burn severity index (ABSI) remained associated with MAKE (OR 1.005 [CI 95% 1.0005-1.009], p=0.03 and OR 1.682 [CI95%1.038-2.726], p=0.035 respectively). Adding pNGAL to abbreviated burn severity index, simplified organ failure assessment and the simplified acute physiology score 2 did outperform clinical scores for the prediction of MAKE and AKI and for most severe forms of AKI and allowed a statistically significant reclassification of patients compared to ABSI for MAKE, RRT, AKI at Day 7 and AKI during hospitalization with a number of patients needed to screen to detect one extra episode of MAKE was 44, 13 for severe AKI and 15 for AKI. CONCLUSIONS: pNGAL at admission is associated with the risk of MAKE in this population, and outperform severity scores when associated. Interventional studies are now needed to assess if impact of biomarkers-guided strategies would improve outcome.

• Klíčová slova
• Acute kidney injury, Burn patients, Major adverse kidney event, Plasmatic Neutrophil Gelatinase-Associated Lipocain,
• MeSH
• akutní poškození ledvin krev   epidemiologie   metabolismus   MeSH
• dospělí MeSH
• hodnocení rizik MeSH
• jednotky intenzivní péče MeSH
• kohortové studie MeSH
• kreatinin metabolismus   MeSH
• kritický stav * MeSH
• lidé středního věku MeSH
• lidé MeSH
• lipokalin-2 krev   MeSH
• mortalita * MeSH
• náhrada funkce ledvin statistika a číselné údaje   MeSH
• obnova funkce * MeSH
• popálení krev   metabolismus   mortalita   MeSH
• prognóza MeSH
• prospektivní studie MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• kreatinin MeSH
• LCN2 protein, human MeSH Prohlížeč
• lipokalin-2 MeSH

Neutrophil gelatinase-associated lipocalin is an extracellular protein produced mostly in kidney. Recently, it has become a promising biomarker of renal damage in vivo. On the other hand, the validation of NGAL as a biomarker for nephrotoxicity estimation in vitro has not been characterized in detail yet. Since the HK-2 cells are frequently used human kidney cell line, we aimed to characterize the production of NGAL in these cells and to evaluate NGAL as a possible marker of cell impairment. We used heavy metals (mercury, cadmium), peroxide, drugs (acetaminophen, gentamicin) and cisplatin to mimic nephrotoxicity. HK-2 cells were incubated with selected compounds for 1-24h and cell viability was measured together with extracellular NGAL production. We proved that HK-2 cells possess a capacity to produce NGAL in amount of 2pg/ml/h. We found a change in cell viability after 24h incubation with all tested toxic compounds. The largest decrease of the viability was detected in mercury, acetaminophen, cisplatin and gentamicin. Unexpectedly, we found also a significant decrease in NGAL production in HK-2 cells treated with these toxins for 24h: to 11±5%, 54±5%, 57±6% and 76±9% respectively, compared with controls (=100%). Our results were followed with qPCR analysis when we found no significant increase in LCN2 gene expression after 24h incubation. We conclude that extracellular NGAL production negatively correlates with HK-2 cell impairment.

• Klíčová slova
• Cell viability, HK-2 cells, In vitro nephrotoxicity, NGAL, Nephrotoxicity markers,
• MeSH
• akutní poškození ledvin chemicky indukované   metabolismus   MeSH
• biologické markery metabolismus   MeSH
• buněčné linie MeSH
• cisplatina toxicita   MeSH
• gentamiciny toxicita   MeSH
• kadmium toxicita   MeSH
• lidé MeSH
• lipokalin-2 genetika   metabolismus   MeSH
• paracetamol toxicita   MeSH
• rtuť toxicita   MeSH
• terc-butylhydroperoxid toxicita   MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• biologické markery MeSH
• cisplatina MeSH
• gentamiciny MeSH
• kadmium MeSH
• LCN2 protein, human MeSH Prohlížeč
• lipokalin-2 MeSH
• paracetamol MeSH
• rtuť MeSH
• terc-butylhydroperoxid MeSH

Recently, using experimental animal model, we demonstrated that porcine buccal pouch mucosal cells reflect increased proliferation capability during primary cultivation in vitro. Although the histological structure and morphogenesis in oral cavity is well recognized, the molecular mechanisms which regulate this process still need further investigation. This study was aimed to analyze the molecular marker expression profile involved in morphogenesis and differentiation capacity of porcine buccal pouch mucosal cells during their long-term primary cultivation in vitro. The experiment was performed on buccal pouch mucosal cells isolated from 80 pubertal crossbred Landrace gilts. After collection, the cells were treated enzymatically and transferred into a primary in vitro culture (IVC) system and cultured for 30 days. The cells were collected for RNA isolation after 7, 15 and 30 days of IVC and were checked for their real-time proliferative status using the RTCA system. We found an increased expression of FN1 and SOX9 genes when calculated against ACTB after 7, and 30 days of IVC, (P less than 0.01, P less than 0.001, respectively). The CXCL12 mRNA was down-regulated after 7, 15 and 30 days of IVC, but not statistically significant. Similar expression profile was observed when calculated against HPRT, however, DAB2 was found to be higher expressed at day 15 of IVC, (P less than 0.05). The cell index measured during real-time cell proliferation was substantially increased between 96 h and 147h of IVC and reached the log phase. Since FN1 and SOX9 revealed significant increase of expression after long-term culture in vitro, it is suggested that expression of these differentiation and stemness genes is accompanied by cell proliferation. Moreover, FN1 and SOX9 might be recognized as new markers of buccal pouch mucosal cell proliferation and differentiation in pigs in in vitro primary culture model.

• MeSH
• buněčná diferenciace MeSH
• chemokin CXCL12 genetika   metabolismus   MeSH
• epitelové buňky cytologie   metabolismus   MeSH
• hypoxanthinfosforibosyltransferasa genetika   metabolismus   MeSH
• lipokalin-2 genetika   metabolismus   MeSH
• messenger RNA genetika   metabolismus   MeSH
• morfogeneze genetika   MeSH
• nádorové supresorové proteiny genetika   metabolismus   MeSH
• prasata MeSH
• primární buněčná kultura MeSH
• proliferace buněk MeSH
• signální transdukce MeSH
• stanovení celkové genové exprese MeSH
• transkripční faktor SOX9 genetika   metabolismus   MeSH
• ústní sliznice cytologie   růst a vývoj   metabolismus   MeSH
• vývojová regulace genové exprese * MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• chemokin CXCL12 MeSH
• hypoxanthinfosforibosyltransferasa MeSH
• lipokalin-2 MeSH
• messenger RNA MeSH
• nádorové supresorové proteiny MeSH
• transkripční faktor SOX9 MeSH