Publications by authors named "Dominika Szoke"

33 Publications

Serum potassium concentrations in COVID-19.

Clin Chim Acta 2021 01 22;512:26-27. Epub 2020 Nov 22.

Clinical Pathology Unit, ASST Fatebenefratelli-Sacco, and Department of Biomedical and Clinical Sciences 'Luigi Sacco', University of Milan, Milano, Italy.

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http://dx.doi.org/10.1016/j.cca.2020.11.012DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7680631PMC
January 2021

Further improvement of the quality of tube transportation system is needed to prevent 'seasonal' pseudohyperkalaemia.

Clin Chim Acta 2020 Nov 14;510:644-646. Epub 2020 Aug 14.

Clinical Pathology Unit, ASST Fatebenefratelli-Sacco, Milan, Italy.

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http://dx.doi.org/10.1016/j.cca.2020.08.021DOI Listing
November 2020

Total laboratory automation: Do stat tests still matter?

Clin Biochem 2017 Jul 5;50(10-11):605-611. Epub 2017 Apr 5.

Clinical Pathology Unit, "Luigi Sacco" University Hospital, Milan, Italy; Department of Biomedical and Clinical Sciences, University of Milan Medical School, Milan, Italy.

During the past decades the healthcare systems have rapidly changed and today hospital care is primarily advocated for critical patients and acute treatments, for which laboratory test results are crucial and need to be always reported in predictably short turnaround time (TAT). Laboratories in the hospital setting can face this challenge by changing their organization from a compartmentalized laboratory department toward a decision making-based laboratory department. This requires the implementation of a core laboratory, that exploits total laboratory automation (TLA) using technological innovation in analytical platforms, track systems and information technology, including middleware, and a number of satellite specialized laboratory sections cooperating with care teams for specific medical conditions. In this laboratory department model, the short TAT for all first-line tests performed by TLA in the core laboratory represents the key paradigm, where no more stat testing is required because all samples are handled in real-time and (auto)validated results dispatched in a time that fulfills clinical needs. To optimally reach this goal, laboratories should be actively involved in managing all the steps covering the total examination process, speeding up also extra-laboratory phases, such sample delivery. Furthermore, to warrant effectiveness and not only efficiency, all the processes, e.g. specimen integrity check, should be managed by middleware through a predefined set of rules defined in light of the clinical governance.
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http://dx.doi.org/10.1016/j.clinbiochem.2017.04.002DOI Listing
July 2017

Frequency of Pancreatic Hyperamylasemia in Human Immunodeficiency Virus-Positive Patients in the Highly Active Antiretroviral Therapy Era.

Am J Clin Pathol 2016 Jan;145(1):128-33

From the Clinical Pathology Unit.

Objectives: Increased frequency of hyperamylasemia has previously been reported in human immunodeficiency virus (HIV)-positive patients, but studies determined total amylase activity and were performed before the introduction of highly active antiretroviral therapy (HAART). We evaluated the frequency of pancreatic hyperamylasemia in a large HIV+ population mostly treated with HAART.

Methods: The upper reference limit (URL) for pancreatic amylase (P-AMY) was derived from 299 healthy blood donors. A cross-sectional study was then performed on samples obtained from 1,548 consecutive patients referred to our infectious disease clinic to assess serum P-AMY and lipase concentrations. Of the patients, 94% were HIV+, and most (92%) were taking HAART (HIV+Tx+).

Results: P-AMY URL was 51 U/L. The frequency of P-AMY increase did not significantly differ between HIV+ and HIV - populations (14.2% vs 15.2%, P = .91) or between HIV+Tx+ and HIV+Tx - (14.7% vs 8.9%, P = .11). In almost half (48.3% of HIV+ and 42.9% of HIV -) of hyperamylasemic patients, lipase was normal, indicating a non pancreatic origin of their P-AMY increase. Markedly elevated P-AMY (>3 times the URL) was found in six HIV+ patients and in one HIV - patient: two had macroamylasemia, one acute pancreatitis, three (including the HIV - patient) chronic pancreatitis, and one chronic hyperamylasemia of undefined origin.

Conclusions: In our study, both HIV+ and HIV+Tx+ do not show an increased frequency of P-AMY elevation. Frank pancreatic disease is rare in this clinical setting.
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http://dx.doi.org/10.1093/ajcp/aqv020DOI Listing
January 2016

Heparinate but not serum tubes are susceptible to hemolysis by pneumatic tube transportation.

Clin Chem Lab Med 2016 May;54(5):785-9

Background: Pneumatic tube transportation (PTT) may induce hemolysis (H) in blood samples. We aimed to compare the H degree before and after PTT implementation in our hospital.

Methods: Hemolysis indices (HI) for all lithium-heparin plasma samples (P) drawn by the Emergency Department in 2-month periods were retrospectively collected and pre- (n=3579) and post-PTT (n=3469) results compared. The impact of PTT introduction was investigated on LDH [HI threshold (HIt), 25], conjugated bilirubin (cBIL) (HIt, 30), K (HIt, 100) and ALT (HIt, 125). In addition, HI retrieved for P and paired serum samples collected in silica clot activator tubes (S) from the same venipuncture were compared in pre- (n=501) and post-PTT (n=509) periods.

Results: Median (5-95th percentile) HI in P was significantly higher in post-PTT period [7 (0-112) vs. 6 (0-82), p<0.001]. Results reported as 'Hemolysis' in P increased from 6.6% in pre-PTT to 9.4% in post-PTT (p<0.001). Investigated tests gave the following rejection rates (pre-PTT vs. post-PTT): LDH, 13.4% vs. 18.8%, p<0.001; cBIL, 9.4% vs. 27.0%, p<0.05; K, 3.7% vs. 5.6%, p<0.001; ALT, 2.9% vs. 4.4%, p<0.01. The slightly higher susceptibility to H of S compared to paired P found in the pre-PTT [9 (1-64) vs. 6 (0-85)] was not confirmed in the post-PTT period [7 (0-90) vs. 8 (1-72)], in which median HI in S was significantly lower (p<0.001) than in pre-PTT.

Conclusions: In our setting PTT promotes H in P, increasing the rate of rejected tests. The use of S appears to protect against the hemolysing effect of PTT.
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http://dx.doi.org/10.1515/cclm-2015-0751DOI Listing
May 2016

The calibrator value assignment protocol of the Abbott enzymatic creatinine assay is inadequate for ensuring suitable quality of serum measurements.

Clin Chim Acta 2015 Oct 11;450:125-6. Epub 2015 Aug 11.

Clinical Pathology Unit, "Luigi Sacco" University Hospital, Milan, Italy; Centre for Metrological Traceability in Laboratory Medicine (CIRME), University of Milan, Milan, Italy.

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http://dx.doi.org/10.1016/j.cca.2015.08.007DOI Listing
October 2015

Measurement of icteric index as approach to detect abnormal total bilirubin values.

J Clin Pathol 2013 Dec 23;66(12):1095-7. Epub 2013 May 23.

Clinical Biochemistry Laboratory, 'Luigi Sacco' University Hospital, and Department of Biomedical and Clinical Sciences, University of Milan Medical School, Milan, Italy.

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http://dx.doi.org/10.1136/jclinpath-2013-201544DOI Listing
December 2013

Fatal electrolyte abnormalities following enema administration.

Clin Chem 2012 Nov;58(11):1515-8

Clinical Biochemistry Laboratory, Luigi Sacco University Hospital, Milan, Italy.

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http://dx.doi.org/10.1373/clinchem.2011.170183DOI Listing
November 2012

Diagnostic value of transferrin.

Clin Chim Acta 2012 Aug 23;413(15-16):1184-9. Epub 2012 Apr 23.

Cattedra di Biochimica Clinica e Biologia Molecolare Clinica, Dipartimento di Scienze Cliniche Luigi Sacco, Università degli Studi, Milano, Italy.

Despite the growing interest in hepcidin and other relatively new biomarkers, guidelines and clinical pathways continue to recommend traditional markers, such as serum transferrin (Tf) and ferritin, as laboratory tests for the diagnostic evaluation of iron-related disorders. In this study, we aimed to critically evaluate the diagnostic role of Tf relying on the highest level of available evidence by a comprehensive literature search. The role of Tf in iron deficiency (ID) and iron overload (IO) syndrome as well as a risk marker was evaluated. The low accuracy of Tf and Tf saturation (TS) in the diagnosis and management of ID conditions does not permit definitively recommending their use, even if recently published guidelines still consider the TS investigation as a complementary test for ferritin. If a tissue IO is suspected, TS is often used, even if it may not be the best test for detecting this condition. Nevertheless, clinical guidelines strongly recommend the use of TS as a first-level test for performing genetic diagnosis of hereditary hemochromatosis. Recently reported data indicating elevated TS as a risk factor for diabetes mellitus, cancer, and total mortality, may provide useful additions to the debate over whether or not to screen for IO using TS.
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http://dx.doi.org/10.1016/j.cca.2012.04.021DOI Listing
August 2012

Implementation of new recommendations for the diagnosis of gestational diabetes: a 5-month audit.

Clin Chem Lab Med 2012 Jul;50(7):1271-3

Clinical Biochemistry Laboratory, Luigi Sacco University Hospital, and Department of Clinical Sciences, University of Milan, Milan, Italy.

Background: Recent international recommendations for the diagnosis of gestational diabetes (GD) were implemented in a university hospital. The aim was to audit the appropriateness of use of the new diagnostic approach.

Methods: The same 5-month period, one before [2009, traditional two-step oral glucose tolerance test (OGTT) approach, S1] and one after the implementation of new criteria (2010, S2) were compared.

Results: In the two periods, 256 (S1) and 245 (S2) pregnant women were examined and 298 (50 g, n = 195; 100 g, n = 103) and 252 (75 g) OGTTs were, respectively, executed. In S1, 54 (27.7% ) 50 g OGTTs resulted positive and 36 (66.7% ) of those performed the 100 g OGTT. In addition, three (1.5% of total) 50 g OGTT negative women were submitted to 100 g OGTT. Sixty-three women did 100 g OGTT only. In total, 14 (13.6% ) 100 g OGTTs were positive. In S2, 38 (15.1% ) 75 g OGTTs were positive. In women who did the complete protocol in the hospital, 98.3% in S1 and 77.0% in S2 performed the correct protocol (p < 0.0001).

Conclusions: In this hospital new recommendations for GD diagnosis are not correctly applied in 23% of cases. The main issue seems to be the lack of consideration of the new threshold for fasting glycemia (5.1 mmol/L) as a main decisional driver for performing OGTT.
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http://dx.doi.org/10.1515/CCLM.2011.757DOI Listing
July 2012

Hemoglobin, bilirubin, and lipid interference on Roche Cobas 6000 assays.

Clin Chim Acta 2012 Jan 5;413(1-2):339-41; author reply 342-3. Epub 2011 Oct 5.

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http://dx.doi.org/10.1016/j.cca.2011.09.044DOI Listing
January 2012

Polymorphisms of the ApoE, HSD3B1, IL-1beta and p53 genes are associated with the development of early uremic complications in diabetic patients: results of a DNA resequencing array study.

Int J Mol Med 2009 Feb;23(2):217-27

Second Department of Internal Medicine, Semmelweis University, Budapest, 1088 Hungary.

Genetic polymorphisms of the genes involved in angiogenesis, the inflammatory cascade or apoptosis control can influence the chronic complications of diabetic patients. Parallel evaluation of multiple genetic polymorphisms became available with the development of DNA resequencing arrays. We aimed to develop a 16-gene, 18,859-nucleotide resequencing array to analyze the genetic background of uremic and gastrointestinal complications. DNA was isolated from 10 ml of peripheral blood of 41 non-uremic and 37 uremic patients with type II diabetes mellitus (DM); 32 suffering from gastric erosion complications. An Affymetrix Customseq Resequencing array was developed containing a total of 37 PCR products of selected genes. Confirmatory analysis was performed for 5 known polymorphisms by RFLP and for 4 others by capillary sequencing. Statistical analysis was performed using the Fisher's exact test. Correlations between the DNA resequencing array and the confirmatory methods were 96% for RFLP and 99.4% for capillary sequencing. The genetic polymorphisms of the ApoE, HSD3B1, IL-1beta and p53 genes were found to be significantly different (p<0.05) between the uremic and non-uremic diabetes group. In regards to the gastric erosion complications of the diabetic uremic patients, the A17708T polymorphism of the p53 intron 10 was found to have a statistically significant (p<0.05) role. In conclusion, DNA sequencing arrays can contribute to a multiparameter genetic analysis yielding highly correlating results using a single method in patients suffering type II DM.
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February 2009

Significantly elevated Helicobacter pylori density and different genotype distribution in erosions as compared with normal gastric biopsy specimen detected by quantitative real-time PCR.

Eur J Gastroenterol Hepatol 2008 Apr;20(4):305-13

Clinical Research Unit, Hungarian Academy of Sciences, Budapest, Hungary.

Background And Aim: Determination of the local densities of Helicobacter pylori and its genotypic variations in gastric biopsy specimens by using novel real-time PCR-based methods could support the precise diagnosis and understanding of H. pylori infections.

Methods: Serial dilutions of H. pylori (0.016-16 microg/microl), control, bacterial, and human DNA samples were prepared. Fresh-frozen gastric biopsy specimens were taken from 103 patients, and the DNA was isolated. Quantitative determination of the ureaseA gene using hybridization probes with parallel evaluation of an internal human control gene (beta-globin) was performed by real-time PCR. CagA and VacA s1 genotypic characterizations were also performed. The data were compared with urea breath test (UBT), histology, and serological testing.

Results: The presence of H. pylori could be detected by ureaseA-fluorescence energy transfer (53%), UBT (51%), serological testing (48%), and histology (52%) when compared with the gold standard (54%). A significant correlation was found between the quantitative real-time ureaseA/beta-globin ratio-based H. pylori frequency and the UBT results (P<0.01). Significantly increased bacterial density was found in the erosions when compared with the healthy part of the antrum and corpus (P<0.01). Real-time PCR VacA s1 results were in significant correlation (P<0.01) with those of serological tests, but CagA results were not. The genomic profiles (VAC/GAC) were different in 13.7% of the cases, which involved three different locations in the stomach.

Conclusion: Real-time PCR was the most reliable method for H. pylori diagnosis. Furthermore, quantification and genotyping could also be performed using this technique. The density of H. pylori was significantly increased in macroscopic erosions.
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http://dx.doi.org/10.1097/MEG.0b013e3282f2fda4DOI Listing
April 2008

Helicobacter pylori and antrum erosion-specific gene expression patterns: the discriminative role of CXCL13 and VCAM1 transcripts.

Helicobacter 2008 Apr;13(2):112-26

2nd Department of Medicine, Semmelweis University, Budapest, Hungary.

Background And Aims: Chronic Helicobacter pylori infection affects approximately half of the world, leads to chronic gastritis and peptic ulceration, and is linked to gastric carcinoma. Our aims were to compare the gene expression profile (GEP) of H. pylori-positive and H. pylori-negative gastric erosions and adjacent mucosa to explain the possible role and response to H. pylori infection and to get erosion-related mRNA expression patterns.

Methods: Total RNA was extracted, amplified, and biotinylated from gastric biopsies of patients with H. pylori-positive and H. pylori-negative antrum erosions (ER) (8/8) and adjacent macroscopically normal mucosae (8/8). The GEP was evaluated using HGU133plus2.0 microarrays. Two independent normalizations (MAS5.0, RMA), PAM feature selection, hierarchical cluster analysis, and discriminant analysis were done. The expression of 14 genes was also measured by real-time-polymerase chain reaction. VCAM-1 and CXCL13 immunohistochemistry (IHC) was done.

Results: In H. pylori infection, significant overexpression of MHC class II antigen-presenting genes, interleukin-7 receptor, ubiquitin-D, CXCR4, lactoferrin immune response-related genes, CXCL-2 and -13, CCL18 chemokine ligand, and VCAM-1 genes were established. In erosive gastritis, increased proliferation (MET) and transport (UCP2, SCFD1, KPNA4) were found, while genes associated with adhesion (SIGLEC11), transcription regulation (ESRRG), and electron and ion transport (ACADM, CLIC6) were down-regulated. Discriminant analysis successfully classified all samples into four groups (HP+ER-, HP+ER+, HP-ER+, HP-ER-) using a reduced gene set (20). Significant overexpression of VCAM-1 and CXC13 protein was detected by IHC in HP+ samples (p < .05).

Conclusions: Whole genomic microarray analysis yielded new H. pylori infection and erosion-related gene expression changes. Discriminative genes can be used in mRNA-based diagnostic classification of gastric biopsies.
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http://dx.doi.org/10.1111/j.1523-5378.2008.00584.xDOI Listing
April 2008

T-251A polymorphism of IL-8 relating to the development of histological gastritis and G-308A polymorphism of TNF-alpha relating to the development of macroscopic erosion.

Eur J Gastroenterol Hepatol 2008 Mar;20(3):191-5

Department of Internal Medicine, Semmelweis University, Budapest, Hungary.

Objectives: Genetic variations of the inflammatory IL-8 and TNF-alpha genes can influence the outcome of gastric alterations. Our aims were to determine the prevalence and effect of the T-251A functional polymorphism of IL-8 and the G-308A polymorphism of TNF-alpha in histological and macroscopic gastric diseases related to Helicobacter pylori infection.

Methods: Genomic DNA was extracted from biopsy samples from patients with gastritis (n=86, H. pylori positive=41), atrophy (n=32, H. pylori positive=13), intestinal metaplasia (IM) (n=43, H. pylori positive=22) and from histologically negative patients (n=57). The samples were divided by macroscopic diagnosis into erosion and negative groups. The T-251A polymorphism was examined with the amplification refractory mutation system method; the G-308A polymorphism was determined by the polymerase chain reaction-restriction fragment length polymorphism method. For statistical evaluation, Fischer's exact test was used.

Results: In the case of T-251A of IL-8, the frequency of the A/A genotype was significantly increased in gastritis (P=0.049) and IM (P=0.038) groups as compared with the histologically negative ones. No relationship was found between macroscopic erosions and H. pylori infection. In the case of G-308A, the G/G genotype frequency was statistically increased in erosions as compared with negative groups (P=0.035). No difference in the distribution of G-308A genotypes in relation to histological alterations and the H. pylori infection was observed.

Conclusions: The effect of the polymorphism of IL-8 seems to be relevant in the pathogenesis of histological gastritis and IM, and the effect of the polymorphism of TNF-alpha is relevant in the pathogenesis of macroscopic erosive gastritis.
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http://dx.doi.org/10.1097/MEG.0b013e3282f1d29fDOI Listing
March 2008

Identification of consensus genes and key regulatory elements in 5-fluorouracil resistance in gastric and colon cancer.

Onkologie 2007 Sep 7;30(8-9):421-6. Epub 2007 Sep 7.

Semmelweis University Budapest, 1st Dept. of Pediatrics, Budapest, Hungary.

Background: 5-fluorouracil (5-FU) is widely used in the treatment of gastric and colorectal cancer. Recent microrarray studies associated different gene lists with 5-FU resistance. A major challenge in the genomic era is to find the most validated genes, and to decipher the regulatory networks responsible for the expression changes in a set of co-regulated transcripts. Our aim was to find genes repeatedly associated with 5-FU resistance, and to identify transcription factors (TFs) having overrepresented binding sites (TFBSs) in the promoter regions of genes associated with 5-FU resistance.

Materials And Methods: The analyzed data originated from 5 different publications describing genome-wide gene expression patterns associated with 5- FU resistance in gastric and colorectal cancer. First, a data warehouse containing all genes associated with resistance was set up. 39 genes were identified which were repeatedly associated with resistance. Of these, using the EZ-Retrieve web service, proximal promoter sequences were available for 33 genes. The MotifScanner software was used to detect TFBSs in this set of sequences.

Results: A total of 200 different TFBSs were identified. Using the statistics tool of the Java program TOUCAN, 4 binding sites were found to be significantly overrepresented: NFKappaB50 (p = 0.01), EGR2 (p = 0.027), EGR3 (p = 0.007), and NGFIC (or EGR4) (p = 0.001). These genes intercept apoptotic pathways at multiple locations in the tumor cells.

Conclusion: We identified a consensus gene list associated with 5-FU resistance, performed an in silico comparative promoter analysis, and highlighted the potential implication of some TFs in the development of chemoresistance.
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http://dx.doi.org/10.1159/000104490DOI Listing
September 2007

Evaluation of malignant and benign gastric biopsy specimens by mRNA expression profile and multivariate statistical methods.

Cytometry B Clin Cytom 2007 Sep;72(5):299-309

2nd Department of Internal Medicine, University Semmelweis, Faculty of Medicine, Budapest, Hungary.

Background: mRNA expression array and multivariate statistical analysis of gastric biopsies can yield insight into the molecular biology basis of local alterations, supporting expression-based identification of morphological alterations.

Methods: From 11 patients with erosive gastritis(EG), 5 with adenocarcinoma (GC), 11 with atrophic gastritis (AG) gastric biopsies were collected, total RNA isolated, T7 amplification and expression analysis of 1047 mRNAs was performed using commercial glass arrays (Clontech, USA). After microarray quality control, applicable data were available from 7 EG, 4 GC, and 5 AG. Multivariate statistical and cell functional analysis were performed. Real-time RT-PCR and immunohistochemistry were used for validation.

Results: GC was characterized by overregulated v-raf, v-erb-a, BCL2-associated- athanogene, immediate-early-response-3, Polo-like kinase, CDK-2, cyclin-C, Pin1 genes, and downregulated ADP-ribosyltransferase, sialophorin and DCC. AG cases had increased PDGF-receptor, TGF-beta-receptor-3, and decreased death-associated-protein-3, beta-1-catenin, topoisomerase-1 levels. In EG upregulation of IGF-receptor-1, CD9, transferrin receptor, integrins, and underexpression of keratin-5, caspase-4 was found. Discriminant analysis could reclassify all samples correctly using four parameters.

Conclusions: mRNA expression array analysis of gastric biopsies yields previously known and new data in the evaluation of local gastric alterations.
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http://dx.doi.org/10.1002/cyto.b.20189DOI Listing
September 2007

Genetic polymorphisms of NOD1 and IL-8, but not polymorphisms of TLR4 genes, are associated with Helicobacter pylori-induced duodenal ulcer and gastritis.

Helicobacter 2007 Apr;12(2):124-31

Department of Medical Microbiology and Immunobiology, University of Szeged, Szeged, Hungary.

Background: Intracellular pathogen receptor NOD1 is involved in the epithelial cell sensing Helicobacter pylori, which results in a considerable interleukin (IL)-8 production. The aim of this study was to evaluate the relationship between NOD1 and IL-8 genetic polymorphisms and the development of H. pylori-induced gastritis and duodenal ulcer (DU), as compared with TLR4 polymorphisms.

Materials And Methods: Eighty-five patients with DU and 135 patients with gastritis were enrolled in the study. Seventy-five serologically H. pylori-positive subjects without gastric or duodenal symptoms served as controls. The G796A (E266K) NOD1 polymorphism was determined by restriction fragment length polymorphism, and the -251 IL-8 polymorphism by amplification refractory mutation system method. The TLR4 (ASP/299/Gly and Thr/399/Ile) gene polymorphisms were examined by melting point analysis.

Results: AA homozygote mutant variants of NOD1 were detected in 20% of the H. pylori-positive patients with DU versus 7% of H. pylori-positive patients with gastritis and versus 6% of the H. pylori-positive healthy controls. The IL-8 heterozygote mutant variant was detected with a significantly higher frequency among the DU patients and those with gastritis than among the H. pylori-positive controls. However, no significant correlation concerning the frequency of the TLR4 gene polymorphism could be revealed between any group of patients and the controls.

Conclusion: E266K CARD4/NOD1, but not the TLR4 gene polymorphism increases the risk of peptic ulceration in H. pylori-positive patients. The -251 IL-8 polymorphism was significantly associated with either gastritis or DU in H. pylori-infected subjects. Host factors including intracellular pathogen receptors and IL-8 production play an important role in H. pylori-induced gastric mucosal damage.
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http://dx.doi.org/10.1111/j.1523-5378.2007.00481.xDOI Listing
April 2007

Laser microdissection in translational and clinical research.

Cytometry A 2006 Sep;69(9):947-60

Second Department of Medicine, Semmelweis University, Budapest, Hungary.

Laser microdissection (LMD) is now a well established method for isolating individual cells or subcellular structures from a heterogeneous cell population. In recent years, cell, DNA, RNA, and protein based techniques has been successfully coupled to LMD and important information has been gathered through the analysis of the genome, transcriptome, and more recently the proteome of individual microdissected cells. The aims of this review are to summarize and compare the principles of different laser microdissection instruments and techniques, to discuss sample preparation procedures for microdissection, and to provide wide variety of examples of translational/clinical research applications of LMD. Novel techniques specifically developed for the improved isolation of stained cells, living cells, or rare cells are also discussed.LMD has become an indispensable tool in the preparation of homogenous samples for sophisticated cell or molecular assays. Despite major technological advances, the labor requirements of LMD are still relatively high. However, understanding the advantages and disadvantages of LMD technology and associated sample preparation procedures may aid in the earlier introduction of this method into the routine clinical diagnostics.
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http://dx.doi.org/10.1002/cyto.a.20322DOI Listing
September 2006

Comparative promoter analysis of doxorubicin resistance-associated genes suggests E47 as a key regulatory element.

Anticancer Res 2006 Jul-Aug;26(4B):2971-6

Second Department of Internal Medicine, Semmelweis University Budapest, Szenkirdlyi u. 46, H-1085, Budapest, Hungary.

Working under the assumption that up- or downregulation of genes implicated in chemoresistance may be the result of altered function of regulatory transcription factors (TF), over-represented TF-binding sites of gene lists previously associated with doxorubicin resistance were the target of our search. First, a data warehouse was set up containing 52 genes which were present in at least two gene lists; of those, proximal promoter sequences (1 kb upstream and 0.05 kb downstream of the transcriptional start sites) could be retrieved from genomic databases for 45 genes using the EZ-Retrieve. The TOUCAN tool MotifScanner, which searches the TRANSFAC database, was used to detect TF-binding sites (TFBSs) in our set of sequences. The statistics tool of the Java program TOUCAN was applied to the data with the appropriate expected frequencies file to compare the measured prevalence to a background model. The most significantly over-represented TFBS was that of E47 (p=0.00024, prevalence: 0.2 vs. background: 8.19E-6). In summary, based on the results of our analysis it is hypothesized that the E47 transcription factor may contribute to doxorubicin resistance.
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September 2006

[The p53 gene and protein in 2005: new results, promising opportunities].

Orv Hetil 2005 Jul;146(30):1587-94

Semmelweis Egyetem, Altalános Orvostudományi Kar, II. Belgyógyászati Klinika, Budapest.

The p53 gene and protein in 2005: new results, promising opportunities. The p53 gene is one of the most important genes which is involved in the regulation of cell division and tumor formation. The normal function of the p53 protein is to arrest the cell division and to turn the cell towards apoptosis in the case of cell stress and DNA damage, thereby to protect the integrity of the genome. Several p53 gene mutations that have function in carcinogenesis have been found in various tumors, including gastrointestinal carcinomas. The loss of p53 response plays an important role in the malignantly transformed cell proliferation. Promising experiments try to substitute the lost functions of the p53 gene. With their help new anti-tumor therapeutic methods should be developed.
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July 2005

[Single cell DNA analysis: possibilities, methods, implications in relation to stem cell therapy].

Orv Hetil 2004 Oct;145(43):2183-90

Semmelweis Egyetem, Altalános Orvostudományi Kar, II. Belgyógyászati Klinika, Budapest.

In the analysis of circulating tumor cells or in the preimplantation genetic diagnosis it is frequently necessary to examine one single cell. Some methods are appropriate to isolate single cells: Laser Assisted Microdissection, magnetic cell separation or FACS. The use of the whole genome amplification methods are needful, because the amount of the DNA extracted from the isolated cells is very low and inappropriate for additional examinations. With different molecular biological methods (e.g. sequencing, chip-technology) it is possible to determine genetic alterations in the analysed cells, and to return the modified cells with in vitro gene technological methods (viral vectors, non viral methods). Our aim is to summarize the methods and the possible technical problems developing during the process of the single cell molecular biological analysis.
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October 2004
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