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1
Content available remote On chromium direct ETAAS determination in serum and urine
100%
EN
A simple, accurate and reliable method for direct electrothermal atomic absorption spectrometric (ETAAS) determination of chromium in serum and urine samples without any preliminary sample pretreatment is described. Instrumental parameters are optimized in order to define: the most suitable atomizer, optimal temperature program and efficient modifier. An appropriate quantification method is proposed taking into account a matrix interference study. Pyrocoated graphite tubes and wall atomization, pretreatment temperature of 700 °C, atomization temperature of 2600 °C, hydrogen peroxide as modifier and standard addition calibration are recommended. The accuracy of the method proposed for Cr determination in serum and urine was confirmed by comparative analysis of parallel samples after wet or dry ashing as well as by the analysis of two certified reference materials: Serum, Clin Rep 1 and Lypochek Urine, level 1. The detection and determination limits achieved for both matrices are 0.08 µg/L and 0.15 µg/L respectively. The relative standard deviation varied between 15 and 18 % for the chromium content in the samples in the range 0.08–0.2 µg/L and between 4 and 7 % for the chromium content in the range 0.2–2.0 µg/L for both matrices. [...]
EN
The calcium-binding protein S100B is produced primarily by astrocytes and exerts concentration-dependent paracrine and autocrine effects on neurons and glia. The numerous findings of a correlation between S100B and traumatic brain injury (TBI) have resulted in the employment of this protein as a clinical biomarker for such injury. Our present aim was to determine whether cycling with (V) or without (NV) vibration alters serum concentrations of S100B. Twelve healthy, male non-smokers (age: 25.3±1.6 yrs, body mass: 74.2±5.9 kg, body height: 181.0±3.7 cm, VO2peak: 56.9±5.1 ml·min-1·kg-1 (means ± SD)) completed in random order two separate trials to exhaustion on a vibrating bicycle (amplitude 4 mm and frequency 20 Hz) connected to an ergometer. The initial workload of 100 W was elevated by 50 W every 5 min and the mean maximal period of exercise was 25:27±1:30 min. The S100B in venous blood taken at rest, immediately after the test, and 30, 60 and 240 min post-exercise exhibited no significant differences (p>0.05), suggesting that cycling with and without vibration does not influence this parameter.
EN
Introduction: Osteoarthritis (OA) is a widely prevalent joint disease leading to motor disability and pain. Appropriate indicators for identifying patients at risk for this progressive disease, identifying molecular events for detecting early phases of the disease, or biomarkers to screen for treatment responses, however, are lacking. Micro RNAs (miRNAs), which play crucial roles in OA, could be potential biomarkers of OA. Because circulating miRNA levels reflect the disease state, they may be useful for OA screening and as diagnostic tools, reducing the need for invasive procedures and minimizing the cost of current diagnostic methods. Materials and methods: The expression levels of 18 microRNAs (let-7e-5p, miR-21-5p, miR-93-5p, miR-101-3p, miR-103a-3p, miR-130a-3p miR-146a-5p, miR-16-5p, miR-193b-3p miR-199a-3p, miR-210-3p, miR-222-3p, miR-22-3p, miR-27a-3p, miR-27b-3p, miR-335-5p, miR-454-3p, and miR-98-5p) were analyzed by quantitative real-time polymerase chain reaction in the cartilage tissues and serum samples of 28 OA patients and were compared to those of 2 healthy controls. Results: Expression of microRNA-146a-5p was significantly upregulated in the cartilage (p=0.006) and serum (p=0.002) of OA patients. The expression levels of miR-146a-5p in the serum were positively correlated with those in the cartilage (Pearson correlation coefficient R=0.32; p=0.002). Conclusion: miR-146a-5p was significantly overexpressed in patients with OA, both in the articular cartilage tissue and serum, with a positive correlation between the levels in both types of samples. Therefore, the miR-146a-5p serum level could reflect the molecular processes in the cartilage, suggesting its clinical utility as a biomarker for OA management. Implementing noninvasive biomarker using serum miRNAs involves the analysis of the misregulated miRNAs linked to the cartilage pathology.
4
Content available remote Serum metallothionein in newly diagnosed patients with childhood solid tumours
88%
EN
Tumour markers are substances produced by malignant cells or by the organism as a response to cancer development. Determination of their levels can, therefore, be used to monitor the risk, presence and prognosis of a cancer disease or to monitor the therapeutic response or early detection of residual disease. Time-consuming imaging methods, examination of cerebrospinal fluid or tumour tissue and assays for hormones and tumour markers have been used for cancer diagnosis. However, no specific marker for diagnosis of childhood solid tumours has been discovered yet. In this study, metallothionein (MT) was evaluated as a prospective marker for such diseases. Serum metallothionein levels of patients with childhood solid tumours were determined using differential pulse voltammetry - Brdicka reaction. A more than 5-fold increase in the amount of metallothionein was found in sera of patients suffering from cancer disease, compared with those in sera of healthy donors. The average metallothionein level in the sera of healthy volunteers was 0.5 ± 0.2 μmol · dm-3 and was significantly different (p<0.05, determined using the Schefe test) from the average MT level found in serum samples of patients suffering from childhood solid tumours (3.4 ± 0.8 μmol · dm-3). Results found in this work indicate that the MT level in blood serum can be considered as a promising marker for diagnostics, prognosis and estimation of therapy efficiency of childhood tumours.
EN
There is growing evidence that proteins are early targets of reactive oxygen species, and that the altered proteins can in turn damage other biomolecules. In this study, we measured the effects of proteins on the oxidation of liposome phospholipid membranes, and the formation of protein hydroperoxides in serum and in cultured cells exposed to radiation-generated hydroxyl free radicals. Lysozyme, which did not affect liposome stability, gave 50% protection when present at 0.3 mg/ml, and virtually completely prevented lipid oxidation at 10 mg/ml. When human blood serum was irradiated, lipids were oxidized only after the destruction of ascorbate. In contrast, peroxidation of proteins proceeded immediately. Protein hydroperoxides were also generated without a lag period in hybrid mouse myeloma cells, while at the same time no lipid peroxides formed. These results are consistent with the theory that, under physiological conditions, lipid membranes are likely to be effectively protected from randomly-generated hydroxyl radicals by proteins, and that protein peroxyl radicals and hydroperoxides may constitute an important hazard to biological systems under oxidative stress.
EN
introduction: The recommended biological material for the parathyroid hormone (PTH) assay is venous blood plasma. According to diagnostic kit manufacturers, the PTH concentration can also be determined in blood serum. The aim of the study was to determine whether differences exist between PTH measured in serum and in plasma. Material and methods: The study was conducted on venous blood samples (plasma and serum) obtained from 92 patients at the Clinic of Endocrinology, University Hospital WAM – CSW. The PTH concentration was measured with an Immulite 1000 analyzer (Siemens), and iPTH kit. Results: The PTH values in plasma and serum were the same only in 3 cases. In 46 samples (50%), a higher hormone level was observed in the plasma, and in 43 cases (47%), a higher concentration was obtained in serum. The absolute differences between the PTH values in the tested biological materials ranged from 0 to 38 pg/mL, and increased together with the hormone concentration. The relative differences (percentage ratio of absolute differences to plasma PTH concentration) ranged from 0 to 34.3%. However, neither the absolute nor the relative differences were statistically significant (p > 0.05). The classification of PTH concentrations according to the reference values for each biological material (i.e. normal, reduced or elevated) was found to be concordant in 83 tested samples (90%). Conclusion: The identified PTH concentrations in the plasma and serum were not identical but comparable and in most cases the differences did not affect the clinical interpretation of the results.
PL
WSTĘP: Materiałem biologicznym zalecanym do oznaczania parathormonu (parathyroid hormone – PTH) jest osocze krwi żylnej pobranej na wersenian sodowo-potasowy, jednak według producentów zestawów diagnostycznych do pomiaru PTH można stosować zarówno osocze, jak i surowicę krwi. Celem pracy było zbadanie, czy wartości PTH oznaczane w surowicy i w osoczu krwi żylnej są porównywalne. MATERIAŁ I METODY: W badaniu wykorzystano próbki krwi żylnej (osocze i surowica) uzyskane od 92 pacjentów Poradni Endokrynologicznej Uniwersyteckiego Szpitala Klinicznego im. Wojskowej Akademii Medycznej – Centralnego Szpitala Weteranów (USK im. WAM – CSW) w Łodzi. Oznaczenia hormonu wykonano na analizatorze Immulite 1000 (Siemens) zestawem iPTH. WYNIKI: Wartości PTH oznaczone w osoczu i w surowicy były jednakowe tylko w 3 przypadkach. W 46 próbkach (50%) wyższe wartości hormonu odnotowano w osoczu, a w 43 przypadkach (47%) w surowicy. Różnice bezwzględne między wartościami PTH w badanych materiałach wahały się od 0 do 38 pg/mL i zwiększały się wraz ze wzrostem stężenia hormonu. Różnice względne (procentowy stosunek różnic bezwzględnych do stężenia PTH w osoczu) wynosiły od 0 do 34,3%. Jednak zarówno różnice bezwzględne, jak i względne nie były znamienne statystycznie (p > 0,05). Po klasyfikacji stężeń – na prawidłowe, obniżone lub podwyższone – na podstawie wartości referencyjnych PTH dla danego materiału biologicznego uzyskano zgodność wyników w 83 badanych próbkach (90%). WNIOSKI: Stężenia PTH oznaczane w osoczu i w surowicy nie są identyczne, ale porównywalne, a różnice w większości przypadków nie wpływają na interpretację kliniczną wyników.
7
Content available remote HPLC study of the pharmacokinetics of K-203
75%
EN
The pharmacokinetics of K-203, a recently synthesized bis-pyridinium aldoxime, have been investigated by reversed-phase HPLC. The K-203 content of serum samples was monitored by HPLC with ultraviolet absorbance detection at 286 nm. The low concentration of K-203 in samples of brain, eyes, and cerebrospinal fluid was also determined by HPLC; quantitative evaluation was possible by use of an amperometric detector.
EN
The potential effect of genetic modification on nutritional properties of potatoes transformed to improve resistance to a necrotic strain of Potato virus Y was determined in a rat experiment. Autoclaved tubers from four transgenic lines were included to a diet in the amount of 40% and compared with the conventional cv. Irga. The experiment lasted 3 weeks and special attention was paid to nutritional properties of diets, caecal metabolism and serum indices. Genetic modification of potato had no negative effect on the chemical composition and nutritional properties of tubers, ecosystem of the caecum, activity of serum enzymes and non-specific defence mechanism of the rats. Obtained results indicate that transgenic potato with improved resistance to PVYN: line R1F (truncated gene coding for PVYN polymerase in sense orientation), R2P (truncated gene coding for PVYN polymerase in antisense orientation), and NTR1.16 (non-translated regions of PVYN genome in sense orientation) are substantial and nutritional equivalence to the non-transgenic cultivar. Tubers of transgenic line NTR2.27 (non-translated regions of PVYN genome in antisense orientation) increased the bulk of caecal digesta and the production of SCFA as compared to tubers of the conventional cultivar and the other transgenic clones. Taking into account some deviations, it seems reasonable to undertake a long-term feeding study to confirm the nutritional properties of tubers of transgenic lines.
EN
Introduction: Beta-glucuronidase (GLU) is a member of the lysosomal glycosidase family that catalyzes hydrolysis of β-D-glucuronic acid residues from the non-reducing end of glycosaminoglycans. Increased activities of GLU have been earlier reported in the serum of alcohol-dependent patients after a chronic heavy drinking period but not after acute intoxication (called binge drinking). The accelerating binge drinking pheno-menon is an alarming public health issue that requires better prevention. Purpose: To determine the activity serum and urinary GLU, after an acute, single, and a large dose of alcohol intoxication. Materials and methods: The serum and urine of eight healthy binge drinkers were collected before binge drinking, and 2 and five days after the drinking session. The activity of GLU was determined by the colorimetric method. Results: There was a tendency to decrease in the serum GLU activity two days after acute alcohol intoxication (binge drinking), which was followed by the significant increase in the GLU activity five days after drinking. The urinary activity of GLU was not changed after intoxication. Conclusion: Alcohol-induced imbalance in the serum GLU activity might be associated with alcohol-induced liver hypoxia and subsequent reperfusion, and can be detected even five days after the drinking session.
EN
Several markers present in tumor tissues and body fluids considerably aid in early diagnosis, enabling both correct decision- making qualifying patients to high-risk groups, monitoring of the patients’ response to treatment implemented and heralding the possibility of distant metastases or local recurrence. The paper presents correlations of elevated serum levels of CYFRA 21-1 and SCC-Ag with clinical stage, histological differentiation grade and survival rates of 89 patients with FIGO stage IB/IIA squamous cell carcinoma of the uterine cervix. Marker levels were assessed prior to surgery using immunoenzymatic assay kits. Two groups of patients were defined: I (n=31; 35%) surviving less than 5 years and II (n=58; 65%) surviving more than 5 years. CYFRA 21-1 levels exceeding 3.5 ng/ml were present in 42% of patients. Elevation of SCC-Ag level above 1.5 ng/ml was seen in 48% of patients. No correlations were noticed between elevation of serum levels of markers studied and clinical stage nor histological differentiation grade. Patients presenting marker levels below reference values benefited from longer survival. Patients surviving less than 5 years had a statistically significant elevation of baseline serum SCC-Ag levels (p<0.001). These results suggest that patients presenting at an early clinical stage of squamous cell carcinoma and having elevated serum levels of CYFRA 21-1 and SCC-Ag, require neoadjuvant chemotherapy prior to surgical treatment.
PL
Obecne w tkance nowotworowej oraz płynach ustrojowych liczne markery wspomagają wczesną diagnostykę, umożliwiają zarówno podjęcie decyzji o zakwalifikowaniu do grupy o wysokim ryzyku zachorowania, jak i monitorowanie odpowiedzi na zastosowane leczenie, ponadto wskazują na możliwość wystąpienia przerzutów odległych lub wznowy miejscowej. W pracy badano związek pomiędzy wzrostem surowiczych stężeń CYFRA 21-1 i SCC-Ag a zaawansowaniem klinicznym, stopniem histologicznego zróżnicowania oraz czasem przeżycia 89 chorych na raka płaskonabłonkowego szyjki macicy w stopniu klinicznego zaawansowania IB i IIA (wg FIGO). Oceny markerów dokonano przed leczeniem operacyjnym za pomocą immunoenzymatycznych zestawów detekcyjnych. Wyłoniono dwie grupy: I – 31 leczonych (35%) z czasem przeżycia <5 lat i II – 58 (65%) z czasem przeżycia >5 lat. Stężenia CYFRA 21-1 wyższe niż 3,5 ng/ml stwierdzono u 42% chorych. Wzrost SCC-Ag powyżej 1,5 ng/ml wykazano u 48% pacjentek. Nie ujawniono zależności pomiędzy wzrostem stężeń badanych markerów a stopniem zaawansowania klinicznego, jak również zróżnicowaniem histologicznym raka. U chorych, u których stwierdzono niższe od wartości referencyjnych stężenia obu markerów, odnotowano dłuższy czas przeżycia. W grupie leczonych z czasem przeżycia krótszym niż 5 lat wykazano istotny statystycznie wzrost wartości wyjściowych stężeń SCC-Ag (p<0,001). Wyniki badań sugerują, że chore na raka płaskonabłonkowego szyjki macicy we wczesnych stadiach zaawansowania klinicznego, u których przed leczeniem odnotowano podwyższone stężenia CYFRA 21-1 i SCC-Ag, wymagają neoadiuwantowej chemioterapii poprzedzającej leczenie operacyjne. Słowa kluczowe: płaskonabłonkowy rak szyjki macicy, CYFRA 21-1, SCC-Ag, surowica
EN
Determination of non-steroidal anti-infl ammatory drugs (NSAIDs) in human and animal physiological fl uids requires the use of appropriate pro-cedures permitting rapid and effi cient recovery of the drugs from biological matrices. The aim of this study was to compare various methods of NSAIDs extraction from human serum that is essential for GC/MS analysis of the drugs. Commercially available biochemistry control human serum supplemented with ibuprofen, paracetamol, fl urbiprofen, nabumeton, naproxen, ketoprofen, mefenamic acid and diclofenac sodium salt was used. Solid phase extraction, HybridSPE- -PPT technology and liquid-liquid extraction were applied for NSAIDs isolation from the serum. GC/MS analysis of the drugs was preceded by their derivatization to trimethylsilyl esters with BSTFA. The highest extraction effi ciency and the best separation of the drugs from endogenous components of the serum were achieved using the solid phase extraction on octadecyl-bonded silica.
PL
Oznaczanie niesteroidowych leków przeciwzapalnych (NLPZ) w płynach fizjologicznych ludzi i zwierząt wymaga zastosowania szybkich i wydajnych procedur izolacji tych leków z biologicznej matrycy. Celem pracy było porównanie różnych metod ekstrakcji NLPZ z surowicy ludzkiej, jako etapu poprzedzającego analizę tych leków techniką GC/MS. W badaniach wykorzystano dostępną handlowo surowicę stosowaną jako materiał kontrolny w oznaczeniach biochemicznych, do której dodano ibuprofen, paracetamol, flurbiprofen, nabumeton, naproksen, ketoprofen, kwas mefenamowy oraz sól sodową diklofenaku. W celu wyizolowania NLPZ z surowicy zastosowano ekstrakcję do fazy stałej, technikę HybridSPE- -PPT oraz ekstrakcję ciecz-ciecz. Leki poddano derywatyzacji do pochodnych trimetylosililowych za pomocą BSTFA, a następnie analizowano techniką GC/MS. Najwyższą wydajność ekstrakcji oraz najlepszy stopień oddzielenia NLPZ od endogennych składników surowicy uzyskano metodą ekstrakcji do fazy stałej w postaci żelu krzemionkowego z aktywnymi grupami oktadecylowymi.
12
Content available remote Top-down peptidomics of bodily fluids
63%
EN
The naturally occurring peptides, mainly arising from the proteolytic cleavage of larger proteins, play several functions within the body (e. g. antihypertensive, immuno-modulatory, anti-microbial and antiviral, mineral carriers). Their presence or the increase of their concentration could be connected to different pathologies and thereby some peptides could be useful biomarkers for the diagnosis or prognosis of the disease. Peptidome research, particularly within biological fluids, therefore represents one of the most interesting and challenging purposes of proteomics. In this review we describe the current state-of-the-art in peptidomics-based studies of several human bodily fluids (serum, plasma, urine, cerebrospinal fluid, saliva, tears, seminal fluid, vitreous humor, pancreatic juice), emphasizing the contribution of top-down proteomic platforms to the deep structural characterization of natural peptides and their posttranslational modifications.
PL
Celem przedstawionych badań było określenie wpływu krioterapii ogólnoustrojowej na zmiany poziomu białek dopełniacza i aktywność bakteriobójczą surowicy krwi. Wykazano wzrost cytolitycznego działania białek dopełniacza surowicy krwi wobec komórek Shigella flexnerii zarówno w trakcie zabiegów jak i po zakończeniu wszystkich serii badań.
EN
The purpose of the study was to investigate the influence of whole-body cryotherapy on the level of proteins of complement and bactericidal activity of serum. Our preliminary result showed that the cytolitic effect of the complement of serum against Shigella flexneri cells significantly increased. These changes seem to be permanent.
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