Publications by authors named "Tomas Zima"

161 Publications

Diagnostic and prognostic value of placental growth factor serum concentration in clear cell renal cell carcinoma.

Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 2021 Feb 22. Epub 2021 Feb 22.

Institute of Medical Biochemistry and Laboratory Diagnostics, General University Hospital and 1.

Background And Aim: Placental Growth Factor (PlGF) plays a crucial role in angiogenesis and was identified as a potential prognostic biomarker in various types of cancer. Therefore, we evaluated the diagnostic accuracy and prognostic value of PlGF serum concentration in patients with clear cell renal cell carcinoma (ccRCC).

Patients And Methods: A total of 49 patients subjected to partial or radical nephrectomy for ccRCC [localized without relapse (lccRCC; n=31), localized with later relapse (rccRCC; n=8), primary metastatic cancer (mccRCC; n=10); median of follow-up 4.4 years] were enrolled in a prospective study to assess the significance of PlGF serum concentration. PlGF was measured prior to surgery and 3 months postoperatively. Our control group consisted of 38 healthy subjects.

Results: PlGF serum concentration was significantly higher in ccRCC compared to controls (P=0.002). The cut-off value of PlGF concentration for the risk of ccRCC was determined at 12.71 pg/mL (AUC=0.729; P=0.0001). Prior to surgery, among ccRCC subgroups, significantly higher PlGF concentration was detected in mccRCC compared to lccRCC (P=0.002). Postoperatively, we observed a tendency to higher PlGF serum concentration in rccRCC compared to lccRCC subgroup, however without significance (P=0.17). The cut-off value for the risk of relapse was 11.41 pg/mL (AUC=0.792; P=0.0003). In subjects with localized ccRCC with PlGF concentration below 11.41 pg/mL 3-years cancer specific survival was 93% compared to 61% in subject with concentration above the cut-off value (P=0.018).

Conclusion: Based on our findings, PlGF serum concentration seems to be a useful biomarker in diagnostics and prediction of prognosis in ccRCC.
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http://dx.doi.org/10.5507/bp.2021.003DOI Listing
February 2021

Avidity of anti-phospholipid antibodies in relation to their levels.

Cent Eur J Immunol 2020 27;45(2):136-143. Epub 2020 Jul 27.

Institute of Medical Biochemistry and Laboratory Diagnostics, First Faculty of Medicine, Charles University and General University Hospital in Prague, Czech Republic.

Introduction: The heterogeneity of anti-phospholipid antibodies can be manifested not only in different antigenic specificities, but also in their avidities. The aim of the study was to investigate the relationship between anti-cardiolipin antibody (aCL) IgG avidities and levels within the range of their titres, from very low to high ones.

Material And Methods: We analyzed 78 serum samples from 60 patients by ELISA with chaotropic agents, using urea concentration of 6 and 8 mol/l and single diluted serum samples. The changes of aCL levels and avidities were explored during a long-term follow-up in 14 patients.

Results: The avidities of aCLs did not differ in the groups of patients classified according to aCL levels. The higher avidity antibodies predominated in our patients and the fluctuation of avidities in the longitudinal follow-up did not show significant differences. No relationship between aCL levels and their avidities was found.

Conclusions: aCL avidities seem to have no relationship with aCL levels and high-avidity aCLs; the potentially deleterious effects might be present also in patients with low and extremely low aCL levels. Avidity of aCLs belongs to stable characteristics with insignificant changes in time.
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http://dx.doi.org/10.5114/ceji.2020.97901DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7792445PMC
July 2020

Mass spectrometry-based proteomic exploration of the small urinary extracellular vesicles in ANCA-associated vasculitis in comparison with total urine.

J Proteomics 2021 Feb 9;233:104067. Epub 2020 Dec 9.

Institute of Pathological Physiology, First Faculty of Medicine, Charles University, Prague, Czech Republic. Electronic address:

ANCA-associated vasculitis (AAV) is a rare, but potentially severe autoimmune disease, even nowadays displaying increased mortality and morbidity. Finding early biomarkers of activity and prognosis is thus very important. Small extracellular vesicles (EVs) isolated from urine can be considered as a non-invasive source of biomarkers. We evaluated several protocols for urinary EV isolation. To eliminate contaminating non-vesicular proteins due to AAV associated proteinuria we used proteinase K treatment. We investigated the differences in proteomes of small EVs of patients with AAV compared to healthy controls by label-free LC-MS/MS. In parallel, we performed an analogous proteomic analysis of urine samples from identical patients. The study results showed significant differences and similarities in both EV and urine proteome, the latter one being highly affected by proteinuria. Using bioinformatics tools we explored differentially changed proteins and their related pathways with a focus on the pathophysiology of AAV. Our findings indicate significant regulation of Golgi enzymes, such as MAN1A1, which can be involved in T cell activation by N-glycans glycosylation and may thus play a key role in pathogenesis and diagnosis of AAV. SIGNIFICANCE: The present study explores for the first time the changes in proteomes of small extracellular vesicles and urine of patients with renal ANCA-associated vasculitis compared to healthy controls by label-free LC-MS/MS. Isolation of vesicles from proteinuric urine samples has been modified to minimize contamination by plasma proteins and to reduce co-isolation of extraluminal proteins. Differentially changed proteins and their related pathways with a role in the pathophysiology of AAV were described and discussed. The results could be helpful for the research of potential biomarkers in renal vasculitis associated with ANCA.
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http://dx.doi.org/10.1016/j.jprot.2020.104067DOI Listing
February 2021

Osteopontin: The Molecular Bridge between Fat and Cardiac-Renal Disorders.

Int J Mol Sci 2020 Aug 4;21(15). Epub 2020 Aug 4.

Department of Biomedical Sciences for Health, Università degli Studi di Milano, 20133 Milan, Italy.

Osteopontin (OPN) is a multifaceted matricellular protein, with well-recognized roles in both the physiological and pathological processes in the body. OPN is expressed in the main organs and cell types, in which it induces different biological actions. During physiological conditioning, OPN acts as both an intracellular protein and soluble excreted cytokine, regulating tissue remodeling and immune-infiltrate in adipose tissue the heart and the kidney. In contrast, the increased expression of OPN has been correlated with the severity of the cardiovascular and renal outcomes associated with obesity. Indeed, OPN expression is at the "cross roads" of visceral fat extension, cardiovascular diseases (CVDs) and renal disorders, in which OPN orchestrates the molecular interactions, leading to chronic low-grade inflammation. The common factor associated with OPN overexpression in adipose, cardiac and renal tissues seems attributable to the concomitant increase in visceral fat size and the increase in infiltrated OPN macrophages. This review underlines the current knowledge on the molecular interactions between obesity and the cardiac-renal disorders ruled by OPN.
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http://dx.doi.org/10.3390/ijms21155568DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7432729PMC
August 2020

A Multicenter Evaluation of a Nongel Mechanical Separator Plasma Blood Collection Tube for Testing of Selected Therapeutic Drugs.

J Appl Lab Med 2020 07;5(4):671-685

BD, Franklin Lakes, NJ.

Background: Some therapeutic drugs are unstable during sample storage in gel tubes. BD Vacutainer® Barricor™ Plasma Blood Collection Tube with nongel separator was compared with plasma gel tubes, BD Vacutainer PST™, PST II, and BD Vacutainer Serum Tube for acetaminophen, salicylate, digoxin, carbamazepine, phenytoin, valproic acid, and vancomycin during sample storage for up to 7 days.

Methods: Seven hospital sites enrolled 705 participants who were taking at least one selected drug. The study tubes were collected and tested at initial time (0 h), after 48 h of storage at room temperature and on day 7 (after additional 5 days of refrigerated storage). The performance of BD Barricor tube was evaluated for each drug by comparing BD Barricor samples with samples from the other tubes at 0 h from the same participant; stability was evaluated by comparing test results from the same tube at 0 h, 48 h, and 7 days.

Results: At 0 h, BD Barricor showed clinically equivalent results for selected therapeutic drugs compared with the other tubes, except phenytoin in BD PST. Phenytoin samples ≥20 µg/mL in BD PST had 10-12% lower values than samples in BD Barricor. During sample storage, all selected drugs remained stable for 7 days in BD Barricor and in serum aliquots. In BD PST, all drugs remained stable except phenytoin and carbamazepine and in BD PST II except for phenytoin.

Conclusion: The BD Barricor Tube is effective for the collection and storage of plasma blood samples for therapeutic drug monitoring without sample aliquoting.
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http://dx.doi.org/10.1093/jalm/jfaa033DOI Listing
July 2020

Alcohol Use in the Czech Republic – Joint Statement of the Society for Addiction Medicine of the J. E. Purkyně Czech Medical Association and the Czech Society of Cardiology

Cent Eur J Public Health 2019 12;27(Suppl):S3-S5

Institute of Medical Biochemistry and Laboratory Diagnostic, First Faculty of Medicine, Charles University and General University Hospital in Prague, Prague, Czech Republic

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http://dx.doi.org/10.21101/cejph.a5972DOI Listing
December 2019

Novel serum markers HSP60, CHI3L1, and IGFBP-2 in metastatic colorectal cancer.

Oncol Lett 2019 Dec 26;18(6):6284-6292. Epub 2019 Sep 26.

Department of Oncology, First Faculty of Medicine, Charles University, and General University Hospital in Prague, 128 08 Prague 2, Czech Republic.

Colorectal cancer (CRC) is the second leading tumor diagnosis in women and men in the Czech Republic. Patient outcome depends on tumor stage at the time of diagnosis and, in metastatic disease, on the localization and extent of distant metastases. The early detection of metastatic liver disease is an important indication for liver surgery. Therefore, novel biomarkers are urgently required. Serum samples were collected from 97 patients with histologically confirmed metastatic CRC at the time of diagnosis or at the time of progression during palliative treatment, and 79 samples from healthy controls. All patients exhibited adequate liver and renal function and signed informed consent was obtained from all patients included in the current study. The serum levels of Heat shock protein 60 (HSP60), Chitinase-3-like protein 1 (CHI3L1) and Insulin-like growth factor binding protein 2 (IGFBP-2) were measured using immunochemistry. The serum levels of HSP60, CHI3L1 and IGFBP-2 were significantly higher in patients with CRC compared with healthy controls. When compared with carcinoembryonic antigen (CEA), HSP60 exhibited the same sensitivity and specificity, while CHI3L1 and IGFBP-2 exhibited decreased sensitivity. Additionally, the serum levels of HSP60 and IGFBP-2 were indicated to be correlated with the presence of liver metastases, which is in contrast to CEA and Cancer antigen 19-9 (CA19-9). Patients with higher HSP60 and IGFBP-2 levels exhibited a significantly worse survival (P<0.001 and 0.007, respectively). The results of the current study indicate HSP60 to be an effective biomarker in patients with metastatic CRC, with it exhibiting an equal sensitivity to CEA. Additionally, HSP60 and IGFBP-2 levels also strongly correlated with extension of liver metastases and exhibited a prognostic value that contrasted that of CEA.
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http://dx.doi.org/10.3892/ol.2019.10925DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6864964PMC
December 2019

Serum levels of TIMP-1 and MMP-7 as potential biomarkers in patients with metastatic colorectal cancer.

Int J Biol Markers 2019 Sep 4;34(3):292-301. Epub 2019 Sep 4.

Department of Oncology, First Faculty of Medicine, Charles University, and General University Hospital in Prague, Czech Republic.

Objective: Tissue inhibitor of metalloproteinases 1 (TIMP-1) and matrix metalloproteinase 7 (MMP-7) were reported to have potent growth promoting activity. Lack of balance between MMPs and TIMPs is an important factor in the development of gastrointestinal malignancies.

Methods: We collected serum samples from 97 patients with metastatic colorectal cancer and 79 samples from healthy controls. Serum levels of TIMP-1 and MMP-7 were measured immunochemically and compared with standard tumor markers carcinoembryonic antigen and CA19-9.

Results: Serum levels of TIMP-1 and MMP-7 were significantly higher in patients with colorectal cancer compared to healthy controls (both, P < 0.001). TIMP-1 and MMP-7 correlate with the presence of colon involvement (P = 0.001; P = 0.012) and the presence of liver metastases (P = 0.002; P = 0.037), and negatively correlate with pulmonary metastases (P = 0.014; P = 0.005). MMP-7 had similar sensitivity and the same specificity as carcinoembryonic antigen. TIMP-1 and MMP-7 had better sensitivity than CA19-9. TIMP-1 and MMP-7 level correlate with worse outcome (P = 0.002).

Conclusion: The results indicate that TIMP-1 and MMP-7 are effective biomarkers in patients with metastatic colorectal cancer with good sensitivity. TIMP-1 and MMP-7 levels strongly correlate with the extent of liver disease and have prognostic value.
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http://dx.doi.org/10.1177/1724600819866202DOI Listing
September 2019

Does the renal expression of Toll-like receptors play a role in patients with IgA nephropathy?

J Nephrol 2020 Apr 5;33(2):307-316. Epub 2019 Sep 5.

Department of Nephrology, 1st Faculty of Medicine and General University Hospital, Charles University, U Nemocnice 2, 128 08, Prague 2, Czech Republic.

The onset of IgA nephropathy (IgAN), characterized by glomerular deposition of IgA-containing immune complexes, is often associated with synpharyngitic hematuria. Innate immune responses mediated by Toll-like receptors (TLR) may play a role in IgAN onset and/or progression. Here, we assessed the expression of TLR 4, 7, 8, and 9 in renal-biopsy specimens from patients with IgAN, with different degree of proteinuria and eGFR, compared with normal-kidney and disease-control tissues (ANCA-associated vasculitis). Renal-biopsy specimens from 34 patients with IgAN and 7 patients with ANCA-associated vasculitis were used. In addition, we used 15 healthy portions of renal-tissue specimens from kidneys after nephrectomy for cancer as control specimens. Expression of TLR 4, 7, 8, and 9 was assessed using immunohistochemical staining of paraffin-embedded renal-biopsy tissue specimens with specific antibodies and evaluated semiquantitatively by light microscopy. Linear discriminant analysis (LDA) was used to test whether intrarenal staining of TLR 4, 7, 8, and 9 distinguished patients with IgAN from controls or correlated with eGFR and/or proteinuria. eGFR was calculated using the creatinine-based formula. Moreover, the biopsies from patients with IgAN were scored according to the Oxford Classification. LDA showed that staining for TLR 4, 7, 8, and 9 was more intense in specimens from IgAN patients compared to normal kidney tissues. The intensity of intrarenal staining of TLRs discriminated four groups of IgAN patients with different eGFR and proteinuria and MEST scoring.
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http://dx.doi.org/10.1007/s40620-019-00640-zDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7170228PMC
April 2020

The standardization of cerebrospinal fluid markers and neuropathological diagnoses brings to light the frequent complexity of concomitant pathology in Alzheimer's disease: The next challenge for biochemical markers?

Clin Biochem 2019 Oct 10;72:15-23. Epub 2019 Jun 10.

Institut de Pathologie Est -Neuropathologie, Hospices civils de Lyon, Lyon, France; University Claude Bernard Lyon 1, Lyon, France; Department of Cancer Cell Plasticity, Cancer Research Centre of Lyon, INSERM U1052, CNRS UMR5286, Lyon, France. Electronic address:

During the last two decades, neuropathological examination of the brain has evolved both technically and scientifically. The increasing use of immunohistochemistry to detect protein aggregates paralleled a better understanding of neuroanatomical progression of protein deposition. As a consequence, an international effort was achieved to standardize hyperphosphorylated-Tau (phospho-TAU), ßAmyloid (Aß), alpha syncuclein (alpha-syn), phosphorylated transactive response DNA-binding protein 43 (phospho-TDP43) and vascular pathology detection. Meanwhile harmonized staging systems emerged in order to increase inter rater reproducibility. Therefore, a refined definition of Alzheimer's disease was recommended., a clearer picture of the neuropathological lesions diversity emerged secondarily to the systematic assessment of concomitant pathology highlighting finally a low rate of pure AD pathology. This brings new challenges to laboratory medicine in the field of cerebrospinal fluid (CSF) markers of Alzheimer's disease: how to further validate total Tau, phospho-TAU, Aß40 and Aß42 and new marker level cut-offs while autopsy rates are declining?
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http://dx.doi.org/10.1016/j.clinbiochem.2019.06.004DOI Listing
October 2019

Matrix Metalloproteinases in Renal Diseases: A Critical Appraisal.

Kidney Blood Press Res 2019 11;44(3):298-330. Epub 2019 Jun 11.

Department of Nephrology, First Faculty of Medicine, Charles University and General University Hospital in Prague, Prague, Czechia.

Matrix metalloproteinases (MMPs) are endopeptidases within the metzincin protein family that not only cleave extracellular matrix (ECM) components, but also process the non-ECM molecules, including various growth factors and their binding proteins. MMPs participate in cell to ECM interactions, and MMPs are known to be involved in cell proliferation mechanisms and most probably apoptosis. These proteinases are grouped into six classes: collagenases, gelatinases, stromelysins, matrilysins, membrane type MMPs, and other MMPs. Various mechanisms regulate the activity of MMPs, inhibition by tissue inhibitors of metalloproteinases being the most important. In the kidney, intrinsic glomerular cells and tubular epithelial cells synthesize several MMPs. The measurement of circulating MMPs can provide valuable information in patients with kidney diseases. They play an important role in many renal diseases, both acute and chronic. This review attempts to summarize the current knowledge of MMPs in the kidney and discusses recent data from patient and animal studies with reference to specific diseases. A better understanding of the MMPs' role in renal remodeling may open the way to new interventions favoring deleterious renal changes in a number of kidney diseases.
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http://dx.doi.org/10.1159/000499876DOI Listing
January 2020

The Significance of Pregnancy-associated Plasma Protein a Serum Concentration in Clear Cell Renal Cell Carcinoma.

Anticancer Res 2019 Jun;39(6):3249-3253

Institute of Medical Biochemistry and Laboratory Diagnostics, General University Hospital and 1st Faculty of Medicine, Charles University, Prague, Czech Republic

Background/aim: Proteinase pregnancy-associated plasma protein A (PAPP-A) modulates the cell growth and carcinogenesis process. Its role in clear cell renal cell carcinoma (ccRCC) remains unclear. This study aimed to evaluate the significance of PAPP-A serum concentration in diagnosis, follow-up and prognosis of ccRCC patients.

Materials And Methods: In a prospective study including 121 patients who underwent radical or partial nephrectomy for ccRCC [localized ccRCC without relapse (n=80), localized ccRCC with later relapse (n=26), primary metastatic cancer (n=15)] PAPP-A serum concentration was assessed preoperatively and in certain subgroups also postoperatively.

Results: PAPP-A serum concentration showed no statistically significant difference between ccRCC and controls and among ccRCC subgroups, respectively. Disease stage and Fuhrman's grade were not shown to affect PAPP-A concentration. The dynamics of postoperative PAPP-A concentrations did not reveal any significance and PAPP-A was not a prognostic factor for cancer related or overall survival.

Conclusion: PAPP-A serum concentration does not seem to be a useful biomarker in ccRCC.
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http://dx.doi.org/10.21873/anticanres.13466DOI Listing
June 2019

Alzheimer's disease: Making the point.

Clin Biochem 2019 Oct 21;72:1-2. Epub 2019 May 21.

Institute of Medical Chemistry and Laboratory Medicine, The First Faculty of Medicine, General University Hospital, U Nemocnice 2 CZ-121 08, Prague 2, Czech republic.

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http://dx.doi.org/10.1016/j.clinbiochem.2019.05.007DOI Listing
October 2019

Galactose-deficient IgA1 and the corresponding IgG autoantibodies predict IgA nephropathy progression.

PLoS One 2019 22;14(2):e0212254. Epub 2019 Feb 22.

General Teaching Hospital, 1st Faculty of Medicine, Charles University, Department of Nephrology, Prague, Czech Republic.

Background: IgA nephropathy (IgAN), the most common primary glomerulonephritis worldwide, has serious outcomes with end-stage renal disease developing in 30-50% of patients. The diagnosis requires renal biopsy. Due to its inherent risks, non-invasive approaches are needed.

Methods: We evaluated 91 Czech patients with biopsy-proven IgAN who were assessed at time of diagnosis for estimated glomerular filtration rate (eGFR), proteinuria, microscopic hematuria, and hypertension, and then followed prospectively. Serum samples collected at diagnosis were analyzed for galactose-deficient IgA1 (Gd-IgA1) using new native-IgA1 and established neuraminidase-treated-IgA1 tests, Gd-IgA1-specific IgG autoantibodies, discriminant analysis and logistic regression model assessed correlations with renal function and Oxford classification (MEST score).

Results: Serum levels of native (P <0.005) and neuraminidase-treated (P <0.005) Gd-IgA1 were associated with the rate of eGFR decline. A higher relative degree of galactose deficiency in native serum IgA1 predicted a faster eGFR decline and poor renal survival (P <0.005). However, Gd-IgA1 has not differentiated patients with low vs. high baseline eGFR. Furthermore, patients with high baseline eGFR that was maintained during follow-up were characterized by low serum levels of Gd-IgA1-specific IgG autoantibodies (P = 0.003).

Conclusions: Including levels of native and neuraminidase-treated Gd-IgA1 and Gd-IgA1-specific autoantibodies at diagnosis may aid in the prognostication of disease progression in Czech patients with IgAN. Future tests will assess utility of these biomarkers in larger patients cohorts from geographically distinct areas.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0212254PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6386256PMC
November 2019

Alcohol Abuse.

Authors:
Tomáš Zima

EJIFCC 2018 Dec 5;29(4):285-289. Epub 2018 Dec 5.

Institute of Medical Biochemistry and Laboratory Diagnostics, First Faculty of Medicine, Charles University and General University Hospital, Prague, Czech Republic.

Chronic alcohol consumption is a world-wide socioeconomic problem. Three metabolic pathways of ethanol were describe in human - alcohol dehydrogenase (ADH), microsomal ethanol oxidizing system (MEOS, CYP2E1) and catalase. Ethanol directly bounds to different molecules (e.g. etylglucuronid) and ethanol per se and its metabolites have toxic effect on biological stuctures. Alcohol abuse is well known for its liver diseases e.g. cirrhosis (the most frequent cause in Europe and US) and hepatocellular carcinoma. Chronic alcohol comsumption leads to cardiovascular diseases (e.g. hypertension, cardiomyopathy), pancreas damage, myopathies, osteoporosis, neurological and psychiatry diseases including fetal alcohol syndrome and addiction. Alcohol consumption may lead to cancer via several mechanisms, per se (solvent for carcinogens) and its metabolites. Acetaldehyde, a cancerogen, has mutagenic effect on DNA, oxidation of ethanol produces the reactive oxygen and nitrogen species with different effects e.g. cell transformation, DNA, protein and lipid damage. The changes of folate metabolism, altered methylation of DNA, reduction of retionic acid influences on cancer development. The high rate of alcohol consumption has become a great social-health problem. Consumption of alcohol is still increasing in many countries, but in some countries is stable or decreasing (e.g. Mediterranean region). The data across Europe shows that 10% of all cancers in men and 3% of all cancers in women can be attributed to alcohol consumption. Australian data suggests that alcohol intake accounts for 5% of the total cancer burden of disease. Alcohol consumption is one of the leading causes of mortality and morbidity in many developed countries.
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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6295593PMC
December 2018

Locomotion in young rats with induced brain cellular edema - effects of recombinant human erythropoietin.

Neuro Endocrinol Lett 2018 Oct;39(4):310-314

Institute of Physiology, First Faculty of Medicine, Charles University in Prague, Prague, Czech Republic.

Objectives: Effect of recombinant human erythropoietin (rhEPO) on spontaneous motor activity was tested in young rats after intraperitoneal (i.p.) administration of rhEPO, followed by induction of cellular brain edema (CE). Induced changes in the spontaneous horizontal locomotor activity was studied by open field test (OFT).

Methods: CE was induced by water intoxication (WI) using standard method of fractional hyperhydration accompanied with desmopressin administration. Using the accepted method of OFT average time spent in locomotion (s) was determined. 48 young rats at the age of 25, and 35 days were divided into three groups - controls, rats after WI (OFT followed after 44 hours), and rats administered with rhEPO prior to application WI (OFT after 48 hours).

Results: In 35-day-old rats rhEPO administration increased the spontaneous locomotor activity, previously decreased by cellular edema. In 25-day-old rats, rhEPO administration prior to the induced CE, decreased spontaneous locomotor activity.

Conclusion: Presented results demonstrate the neuroprotective capacity of rhEPO, manifested by elimination of the suppressive influence of CE on the locomotion in 35-day-old rats. In 25-day-old rats the neuroprotective effect was not present. These results confirmed that the 10 day interval in the development may represent a different stage of brain maturation in the relation to the neuroprotective effect of rhEPO.
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October 2018

Gene Expression Analysis of Immunomagnetically Enriched Circulating Tumor Cell Fraction in Castration-Resistant Prostate Cancer.

Mol Diagn Ther 2018 06;22(3):381-390

Institute of Medical Biochemistry and Laboratory Diagnostics, General University Hospital and First Faculty of Medicine, Charles University in Prague, U Nemocnice 2, 12808, Prague, Czech Republic.

Background: Molecular characterization of tumors could be a key to therapeutic decision-making with regards to targeted therapies in castration-resistant prostate cancer (CRPC). A convenient solution may be non-invasive liquid biopsy testing of circulating tumor cells (CTCs). For this reason, CTC-enriched samples obtained by immunomagnetic separation (AdnaTest) were studied as a source material for high-throughput gene expression analysis using BioMark™.

Patients And Methods: CTC-enriched samples from 41 CRPC patients previously determined to be CTC positive using the AdnaTest were retrospectively re-analysed for androgen receptor (AR) messenger RNA (mRNA), using the updated AdnaTest. Blood samples were drawn two times from each patient: at the time of CRPC diagnosis and after the third docetaxel cycle. A gene expression panel of 27 genes related to CRPC therapeutic decision-making, including AR full length (ARFL) and splice variant 7 (ARV7), was retrospectively analyzed on a BioMark™ platform in 29 of 41 patients.

Results: The AdnaTest detected AR mRNA in three-quarters of CTC-positive samples taken at the time of CRPC diagnosis and after the third docetaxel cycle. AR detection was associated with a shorter disease-specific survival (45.0 vs. 20.4 months) at the time of CRPC diagnosis. ARFL expression at the time of CRPC diagnosis, measured on the BioMark™ platform, was associated with a lower decrease of serum level of prostate-specific antigen (sPSA) (p = 0.029), i.e., worse therapy response. ARV7 was found in 38% of the ARFL--positive samples at both analyzed timepoints.

Conclusion: Detection of AR expression by AdnaTest in CTC-enriched samples may help predict patients' survival. These AdnaTest CTC-enriched samples can be used in a high-throughput quantitative polymerase chain reaction (qPCR) analysis of gene expression, provided that the specificity of the assay for each individual gene is properly validated. The BioMark™ platform can be used for the simultaneous detection of ARFL and ARV7 and other genes in CTC-enriched samples from CRPC patients.
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http://dx.doi.org/10.1007/s40291-018-0333-0DOI Listing
June 2018

Importance of the integrated test in the Down's syndrome screening algorithm.

J Med Screen 2018 09 25;25(3):114-118. Epub 2018 Mar 25.

1 First Faculty of Medicine, Department of Medical Biochemistry and Laboratory Diagnostics, Charles University, Prague, Czech Republic.

Objective: In the Czech Republic, over 97% of all pregnant women undergo some type of antenatal screening for Down's syndrome. In about 95% of cases with a confirmed fetal chromosomal abnormality, the pregnancy is terminated. The most commonly used test is the first trimester combined test. We investigated the impact of implementing an integrated sequential test to improve the detection of Down's syndrome pregnancies.

Methods: Data on the incidence of congenital defects, number of births, and affected pregnancies terminated are recorded in the National Registry of Congenital Anomalies. Anonymous data on cases of Down's syndrome diagnosed antenatally or postnatally between 2010 and 2015 in one of the large antenatal care centers were analyzed.

Results: There were 600 diagnoses of Down's syndrome (5.7 per 1000 births), 90% of which were made antenatally. Of antenatally detected cases, 80% were indicated for diagnostic procedure by multimarker screening results. In the multimarker screen positive group, 75% cases were first trimester positive and 25% second trimester positive (most of these had positive integrated test results). Among Down's syndrome cases indicated for antenatal diagnosis by multimarker screening results 6.25% (n = 26) were first trimester negative, and became positive after integration with the second trimester screening results.

Conclusions: Results from five major Czech antenatal centers confirm that an integrated sequential test would detect 80-85% of Down's syndrome fetuses in the first trimester and at least an extra 5-10% of Down's syndrome pregnancies in the second trimester of pregnancy. These are important data that should be considered in implementing the national antenatal screening program.
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http://dx.doi.org/10.1177/0969141317752533DOI Listing
September 2018

Growth/differentiation factor 15 (GDF-15) as new potential serum marker in patients with metastatic colorectal cancer.

Cancer Biomark 2018 ;21(4):869-874

Department of Oncology, First Faculty of Medicine, Charles University, and General University Hospital in Prague, Prague 2, 128 08, Czech Republic.

Background: GDF-15 is a protein belonging to the transforming growth factor beta superfamily that has a role in regulating inflammatory and apoptotic pathways. High level GDF-15 in tumor tissues and plasma correlate with an increased risk of recurrence and reduced overall survival.

Objective: The aim of this study was to screen GDF-15 capacity to detecting metastatic CRC and compare it with standard tumor markers CEA and CA19-9.

Methods: We collected serum samples from 97 patients with metastatic colorectal cancer and 79 samples from healthy controls. Serum levels of GDF-15, CEA and CA19-9 were measured by immunochemically. A Kaplan-Meier curve was applied for analysis of survival rates, and a log-rank was used for univariate analysis.

Results: Serum levels of GDF-15 were significantly higher in patients with colorectal cancer compared to healthy controls (p< 0.001). In addition, serum levels of GDF-15 correlated with extent of liver involvement and patients with higher GDF-15 levels had significantly worse outcome (p< 0.0001).

Conclusions: Our results show GDF-15 as an effective biomarker in patients with metastatic colorectal cancer with the same sensitivity as CEA. In addition, GDF-15 levels strongly correlate with extension of liver involvement in contrast with CEA.
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http://dx.doi.org/10.3233/CBM-170792DOI Listing
August 2018

Placental Growth Factor in Bladder Cancer Compared to the Diagnostic Accuracy and Prognostic Performance of Vascular Endothelial Growth Factor A.

Anticancer Res 2018 01;38(1):239-246

Institute of Medical Biochemistry and Laboratory Diagnostics, the First Faculty of Medicine, Charles University and General University Hospital, Prague, Czech Republic.

Background/aim: To evaluate the diagnostic accuracy and prognostic performance of urinary and plasma levels of placental growth factor (PLGF) and provide their comparison with the results of vascular endothelial growth factor A (VEGF-A) in patients with primary and recurrent urinary bladder cancer.

Materials And Methods: The enzyme-linked immunosorbent assay (ELISA) was used to assess urinary and plasma concentrations of PLGF and VEGF-A in 240 individuals.

Results: PLGF levels in urine and plasma were significantly higher in patients with primary bladder cancer than in healthy individuals (p=0.003, p=0.005, respectively). Area under the curve (AUC) of urinary PLGF was 0.68; AUC of plasma PLGF levels was 0.65. Patients with the urine levels of PLGF higher than 82.33 pg/ml had three times higher risk of recurrence. In patients with recurrent bladder cancer, the urinary concentrations of PLGF did not significantly differ from the concentrations in patients without current disease (p=0.61). However, plasma PLGF levels were significantly higher in patients diagnosed with tumor recurrence (p=0.001); AUC of plasma PLGF levels was 0.69. Moreover, patients with plasma levels higher than 10.09 pg/ml had a five-times higher risk of future tumor recurrence. The diagnostic accuracy of PLGF was comparable with VEGF-A.

Conclusion: From a clinical point of view, PLGF could be considered a valid diagnostic test for the detection of primary and recurrent bladder cancer. In patients with recurrent bladder cancer, plasma PLGF levels can differentiate individuals at risk of tumor recurrence.
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http://dx.doi.org/10.21873/anticanres.12213DOI Listing
January 2018

Diagnostic and prognostic value of presepsin vs. established biomarkers in critically ill patients with sepsis or systemic inflammatory response syndrome.

Clin Chem Lab Med 2018 03;56(4):658-668

Department of Anesthesiology, University of Pittsburgh School of Medicine, UPMC Presbyterian Hospital, 200 Lothrop St. Suite C220, Pittsburgh, PA 15213, USA.

Background: Inflammatory biomarkers may aid to distinguish between systemic inflammatory response syndrome (SIRS) vs. sepsis. We tested the hypotheses that (1) presepsin, a novel biomarker, can distinguish between SIRS and sepsis, and (2) higher presepsin levels will be associated with increased severity of illness and (3) with 28-day mortality, outperforming traditional biomarkers.

Methods: Procalcitonin (PCT), C-reactive protein (CRP), presepsin, and lactate were analyzed in 60 consecutive patients (sepsis and SIRS, n=30 per group) on day 1 (D1) to D3 (onset sepsis, or after cardiac surgery). The systemic organ failure assessment (SOFA) score was determined daily.

Results: There was no difference in mortality in sepsis vs. SIRS (12/30 vs. 8/30). Patients with sepsis had higher SOFA score vs. patients with SIRS (11±4 vs. 8±5; p=0.023), higher presepsin (AUC=0.674; p<0.021), PCT (AUC=0.791; p<0.001), CRP (AUC=0.903; p<0.0001), but not lactate (AUC=0.506; p=0.941). Unlike other biomarkers, presepsin did not correlate with SOFA on D1. All biomarkers were associated with mortality on D1: presepsin (AUC=0.734; p=0.0006; best cutoff=1843 pg/mL), PCT (AUC=0.844; p<0.0001), CRP (AUC=0.701; p=0.0048), and lactate (AUC=0.778; p<0.0001). Multiple regression analyses showed independent associations of CRP with diagnosis of sepsis, and CRP and lactate with mortality. Increased neutrophils (p=0.002) and decreased lymphocytes (p=0.007) and monocytes (p=0.046) were also associated with mortality.

Conclusions: Presepsin did not outperform traditional sepsis biomarkers in diagnosing sepsis from SIRS and in prognostication of mortality in critically ill patients. Presepsin may have a limited adjunct value for both diagnosis and an early risk stratification, performing independently of clinical illness severity.
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http://dx.doi.org/10.1515/cclm-2017-0839DOI Listing
March 2018

Fecal zonulin is elevated in Crohn's disease and in cigarette smokers.

Pract Lab Med 2017 Dec 23;9:39-44. Epub 2017 Sep 23.

Institute of Medical Biochemistry and Laboratory Diagnostics, General University Hospital & 1st Faculty of Medicine of the Charles University, U Nemocnice 2, Prague 2 12800, Czech Republic.

Objectives: Human zonulin is a protein that increases permeability in the epithelial layer of the small intestine by reversibly modulating the intercellular tight junctions. There is not sufficient information available about zonulin's participation in inflammatory bowel diseases (IBD). The aim of this study was therefore to investigate fecal and serum zonulin in IBD patients and its relation to the disease localization, behavior and smoking status.

Design And Methods: Forty IBD patients and forty healthy persons were examined for fecal and serum zonulin concentrations by competitive ELISA (DRG International Inc). Values were correlated to IBD type, localization and behavior, and smoking.

Results: Serum and fecal zonulin were significantly higher in patients with Crohn's disease compared to ulcerative colitis (p = 0.038 for fecal zonulin, and p = 0.041 for serum zonulin concentrations). No association of serum or fecal zonulin was found with respect to IBD localization and behavior. The only difference was found with respect to smoking. Both the IBD cohort and healthy smokers showed significantly higher fecal zonulin levels (median 203 ng/mL) compared to non-smokers (median 35.8 ng/mL), p < 0.001.

Conclusions: Fecal and serum zonulin levels are elevated in patients with active Crohn's disease but not with ulcerative colitis. High fecal zonulin levels in smokers irrespective of IBD point to the significant and undesirable up-regulation of gut permeability in cigarette smokers.
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http://dx.doi.org/10.1016/j.plabm.2017.09.001DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5633835PMC
December 2017

Pregnancy-Associated Plasma Protein A2 in Hemodialysis Patients: Significance for Prognosis.

Kidney Blood Press Res 2017 30;42(3):509-518. Epub 2017 Aug 30.

Institute of Medical Biochemistry and Laboratory Diagnostics, First Faculty of Medicine, Charles University and General University Hospital in Prague, Prague, Czech Republic.

Background: Pregnancy-associated plasma protein A (PAPP-A) is associated with adverse outcome of long-term hemodialysis patients (HD). The aim of the study was to test whether its homolog pregnancy-associated plasma protein A2 (PAPP-A2) can be detected in serum of HD patients and to define its significance.

Methods: The studied group consisted of 102 long-term HD patients and 25 healthy controls. HD patients were prospectively followed up for five years (2009-2014). PAPP-A2 was measured by surface plasmon resonance biosensor, PAPP-A by time resolved amplified cryptate emission.

Results: PAPP-A2, similarly as PAPP-A, was significantly increased in HD patients (median (interquartile range)) PAPP-A2: 6.2 (2.6-10.8) ng/mL, vs. 3.0 (0.7-5.9) ng/mL, p=0.006; PAPP-A: 18.9 (14.3-23.4) mIU/L, vs. 9.5 (8.4-10.5) mIU/L, p<0.001). In HD patients, PAPP-A2 correlated weakly but significantly with PAPP-A (τ=0.193, p=0.004). Unlike PAPP-A, PAPP-A2 was not significant for prognosis of HD patients when tested alone. There was a significant interaction between PAPP-A and PAPP-A2 on the mortality due to infection of HD patients (p=0.008). If PAPP-A was below median, mortality due to infection was significantly higher for patients with PAPP-A2 values above median than for patients with low PAPP-A2 levels (p=0.011).

Conclusion: PAPP-A2 is increased in HD patients and interacts with PAPP-A on patients´ prognosis.
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http://dx.doi.org/10.1159/000479847DOI Listing
June 2018

Accreditation of Medical Laboratories - System, Process, Benefits for Labs.

Authors:
Tomáš Zima

J Med Biochem 2017 Sep 14;36(3):231-237. Epub 2017 Jul 14.

Institute of Medical Biochemistry and Laboratory Diagnostics, First Faculty of Medicine, Charles University, General University Hospital Prague, Czech Republic.

One and key of the priorities in laboratory medicine is improvement of quality management system for patient safety. Quality in the health care is tightly connected to the level of excellence of the health care provided in relation to the current level of knowledge and technical development. Accreditation is an effective way to demonstrate competence of the laboratory, a tool to recognize laboratories world-wide, is linked to periodical audits, to stimulate the maintenance and improvement of the quality, which leads to high standard of services for clients (patients, health care providers, etc.). The strategic plans of IFCC and EFLM include focusing on accreditation of labs based on ISO standards and cooperation with European Accreditation and national accreditation bodies. IFCC and EFLM recognised that ISO 15189:2012 Medical laboratories - Requirements for quality and competence, encompasses all the assessment criteria specified in the policy of quality. The last version is oriented to process approach with detailed division and clearly defined requirements. The accreditation of labs improves facilitation of accurate and rapid diagnostics, efficiency of treatment and reduction of errors in the laboratory process. Accreditation is not about who the best is, but who has a system of standard procedures with aim to improve the quality and patient safety. Quality system is about people, with people and for people.
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http://dx.doi.org/10.1515/jomb-2017-0025DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6287213PMC
September 2017

Polymorphisms of the receptor for advanced glycation end products and glyoxalase I and long-term outcome in patients with breast cancer.

Tumour Biol 2017 Jul;39(7):1010428317702902

2 Institute of Medical Biochemistry and Laboratory Diagnostics, First Faculty of Medicine, Charles University and General University Hospital, Prague, Czech Republic.

Receptor for advanced glycation end products and glyoxalase I metabolizing advanced glycation end product precursors may play important role in the pathogenesis and progression of cancer. Potential relation between soluble forms of receptor for advanced glycation end products (sRAGE), receptor for advanced glycation end products, glyoxalase I polymorphisms, and long-term outcome (median follow-up of 10.3 years) was studied in 116 patients with breast cancer. Gly82Ser and 2184 A/G RAGE polymorphisms were related to the mortality due to the breast cancer and -419 A/C glyoxalase I polymorphism was related to the overall mortality of the patients suggesting their role not only in the risk of breast cancer but also in the outcome of patients with breast cancer.
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http://dx.doi.org/10.1177/1010428317702902DOI Listing
July 2017

Prognostic Importance of Vitamins A, E and Retinol-binding Protein 4 in Renal Cell Carcinoma Patients.

Anticancer Res 2017 07;37(7):3801-3806

Department of Urology, General University Hospital and 1st Faculty of Medicine, Charles University, Prague, Czech Republic

Aim: To assess the prognostic importance of serum levels of retinol, retinol-binding protein 4 (RBP4) and vitamin E at the time of diagnosis in patients with renal cell carcinoma (RCC).

Patients And Methods: In this prospective study, in a cohort of 102 renal cell carcinoma patients, relationships between serum levels of the aforementioned markers and recurrence-free survival (RFS), overall survival (OS), as well as cancer-specific survival (CSS), were evaluated. The vitamin A and vitamin E levels were determined by high-performance liquid chromatography (HPLC), while the RBP4 level by enzyme-linked immunosorbent assay (ELISA).

Results: The median follow-up period was 39 months. Renal cell carcinoma recurred in 9 patients; 23 patients died with 12 of them from RCC. The preoperative vitamin E level was associated to RFS (p=0.02). We found a significant relationship between OS and the level of RBP4 (p=0.002), retinol (p=0.037) and vitamin E (p=0.007). The CSS period was significantly associated with the level of RBP4 (p=0.0001) and retinol (p=0.0003). Patients with an RBP4 level less than 21.0 mg/l at the time of diagnosis had a 13.5-times higher risk of death due to RCC progression; this risk was up to 7.7-times higher with vitamin A levels under 0.52 mg/l.

Conclusion: Low levels of vitamin A, E and RBP4 at the time of RCC diagnosis are associated with a poorer prognosis after surgery.
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http://dx.doi.org/10.21873/anticanres.11757DOI Listing
July 2017

Circulating tumor cells and serum levels of MMP-2, MMP-9 and VEGF as markers of the metastatic process in patients with high risk of metastatic progression.

Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 2017 Sep 16;161(3):272-280. Epub 2017 May 16.

Department of Oncology, General University Hospital in Prague and First Faculty of Medicine, Charles University, Prague, Czech Republic.

Background And Aims: Metastases are a severe complication in cancer patients and biomarkers predicting their progression are still lacking for specific groups of patients. HER2 positive breast cancer (HER2 BC) patients on trastuzumab therapy are at risk of the development of unpredictable and often fatal central nervous system (CNS) metastases and castration resistant prostate cancer (CRPC) patients urgently need a marker of disease progression during therapy. Proposed metastatic markers: circulating tumor cells (CTC), serum levels of matrix metalloproteinase 2 (MMP-2), 9 (MMP-9) and vascular endothelial growth factor (VEGF) were prospectively studied to confirm their utility in these two narrowly defined groups of cancer patients.

Patients And Methods: The groups comprised 44 advanced HER2 BC, 24 CRPC patients and 42 healthy controls. An immunomagnetic separation method followed by PCR and electrophoretic detection (AdnaGen, Germany) were used for CTC determination. Serum marker levels were determined by the ELISAs (R&D System, USA).

Results: MMP-2 serum level was significantly higher in HER2 BC patients who developed CNS metastases, especially if there were also bone metastases. CTCs were a negative predictive marker for overall survival in HER2 BC patients. MMP-9 serum level was significantly higher in CRPC patients in whom disease progression occurred. CTC vanished from the blood of most of the CRPC patients (from 88% to 37%) during chemotherapy.

Conclusion: MMP-2 serum level and CTCs show the potential to predict CNS metastases and overall survival in BC patients. CTCs and MMP-9 serum level could be a promising therapy response marker in CRPC patients.
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http://dx.doi.org/10.5507/bp.2017.022DOI Listing
September 2017

Rivaroxaban - Metabolism, Pharmacologic Properties and Drug Interactions.

Curr Drug Metab 2017 ;18(7):636-642

Thrombotic Centre of Institute of Medical Biochemistry and Laboratory Diagnostics, General University Hospital, Karlovo Namesti 32, 121 11 Prague 2. Czech Republic.

Background: Rivaroxaban represents a selective direct inhibitor of activated coagulation factor X (FXa) having peroral bioavailability and prompt onset of action.

Objective: The absorbtion of rivaroxaban is quick, reaching maximum plasma concentration 2-4 hours following its administration. Peroral bioavailability is high (80-100 %) and pharmacokinetic variability is considered to be moderate (coefficient of variation 30-40 %). This review discusses the properties, drug interactions, pharmacokinetics and clinical indications of rivaroxaban.

Method: Dosing regimen of rivaroxaban was derived from pharmacologic data of the development program aimed to gain strong antithrombotic drug and balance between efficacy and risk of bleeding in patients. Results of doseranging trials, pharmacokinetic models and randomised studies of phase III advocate the use of such schemes in everyday practice.

Results: The drug has been manufactured to fulfill clinical requirements in a variety of indications in adults: prophylaxis of venous thromboembolism (VTE) following elective knee or hip replacement surgical intervention, therapy and secondary prophylaxis of VTE, prophylaxis of ischemic stroke and embolism in individuals diagnosed with nonvalvular atrial fibrillation (NVAF) with risky characteristics, and in Europe the prophylaxis of atherothrombotic episodes following an acute coronary syndrome in subjects with increased levels of cardiac biomarkers.

Conclusion: Rivaroxaban may offer benefit in many clinical situations. In comparison with low molecular weight heparin and fondaparinux requiring subcutaneous way of administration, and with vitamin K antagonists (VKAs), which require regular monitoring of international normalized ratio, rivaroxaban is relatively easy to use. However, adjustments of dose are needed in individuals with impaired renal functions.
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http://dx.doi.org/10.2174/1389200218666170518165443DOI Listing
September 2018