[111In]In-CP04 as a novel cholecystokinin-2 receptor ligand with theranostic potential in patients with progressive or metastatic medullary thyroid cancer: final results of a GRAN-T-MTC Phase I clinical trial.

INTRODUCTION: Medullary thyroid cancer (MTC) is a rare malignant tumour of the parafollicular C-cells with an unpredictable clinical course and currently suboptimal diagnostic and therapeutic options, in particular in advanced disease. Overexpression of cholecystokinin-2 receptors (CCK2R) represents a promising avenue to diagnostic imaging and targeted therapy, ideally through a theranostic approach. MATERIALS AND METHODS: A translational study (GRAN-T-MTC) conducted through a Phase I multicentre clinical trial of the indium-111 labelled CP04 ([111In]In-CP04), a CCK2R-seeking ligand was initiated with the goal of developing a theranostic compound. Patients with proven advanced/metastatic MTC or short calcitonin doubling time were enrolled. A two-step concept was developed through the use of low- and high-peptide mass (10 and 50 µg, respectively) for safety assessment, with the higher peptide mass considered appropriate for therapeutic application. Gelofusine was co-infused in a randomized fashion in the second step for the evaluation of potential reduction of the absorbed dose to the kidneys. Imaging for the purpose of biodistribution, dosimetry evaluation, and diagnostic assessment were performed as well as pre-, peri-, and postprocedural clinical and biochemical assessment. RESULTS: Sixteen patients were enrolled. No serious adverse events after application of the compound at both peptide amounts were witnessed; transient tachycardia and flushing were observed in two patients. No changes in biochemistry and clinical status were observed on follow-up. Preliminary dosimetry assessment revealed the highest dose to urinary bladder, followed by the kidneys and stomach wall. The effective dose for 200 MBq of [111In]In-CP04 was estimated at 7±3 mSv and 7±1 mSv for 10 µg and 50 µg CP04, respectively. Administration of Gelofusine reduced the dose to the kidneys by 53%, resulting in the organ absorbed dose of 0.044±0.019 mSv/MBq. Projected absorbed dose to the kidneys with the use of [177Lu]Lu-CP04 was estimated at 0.9±0.4 Gy/7.4 GBq. [111In]In-CP04 scintigraphy was positive in 13 patients (detection rate of 81%) with superior diagnostic performance over conventional imaging. CONCLUSION: In the present study, [111In]In-CP04 was shown to be a safe and effective radiopharmaceutical with promising theranostic characteristics for patients with advanced MTC.

Gene expression with corresponding pathways analysis in Gaucher disease.

Gaucher disease (GD) caused by mutation in the GBA gene has a wide spectrum of phenotypes. Besides the storage disorder, secondary alteration of various pathways occurs with modification of the expression of many genes. In our work we analysed the expression profile of genes in adult patients with type 1 GD. METHODS: This study was an observational, cross-sectional analysis of a group of twenty patients with type 1 GD and ten healthy volunteers as a control group. First, on the group of ten persons, microarray gene analysis was performed. Afterwards, significantly regulated genes were selected, and the microarray results were confirmed by real-time PCR on the whole study group. RESULTS: Based on the microarray results in the pathway analysis, we focused on genes related to chemokines, inflammatory processes, endocytosis, autophagy, and apoptosis. Patients with GD demonstrated up-regulation of genes related to NFkB pathway (NFkB, NKkBR SQSTM1), inflammation (IL-1b), endocytosis and autophagy (BCN1, SMAD), genes coding proteins involved in apoptosis (CASP, NFkB, BCL2) as well as genes related to proteasome degradation (PSMD2, PSMB9) and SNARE complex (SNAP, STX). Simultaneously, we showed down-regulation of genes coding proteins of chemokines and their receptors (GNB4, CCL5). The qRT-PCR results confirmed changes in expression of selected genes. Parallel microarray results showed inhibition of genes related to neurones development and survival (NTRK1) and stimulation of gene expression related to neurodegeneration and apoptosis (BCN1, IL1B). CONCLUSIONS: The work revealed different pathway activation, especially inflammatory processes followed by autophagy and apoptosis. Our results also pay attention to new pathways leading to disorders of the functioning of the nervous tissue in patients with type 1 GD, which may lead to the development of polyneuropathy and chronic pain. These are clinical symptoms that severely decrease the quality of life in GD patients.

A proposed Common Training Framework for Specialists in Laboratory Medicine under EU Directive 2013/55/EC (The Recognition of Professional Qualifications).

European Union (EU) Directive 2013/55/EC (The Recognition of Professional Qualifications) allows Member States to decide on a common set of minimum knowledge, skills and competences that are needed to pursue a given profession through a Common Training Framework. To be adopted the framework must combine the knowledge, skills and competences of at least one third of the Member States. Professionals who have gained their qualifications under a Common Training Framework will be able to have these recognised automatically within the Union. The backbone of the European Federation of Clinical Chemistry and Laboratory Medicine's (EFLM) proposed Common Training Framework for non-medical Specialists in Laboratory Medicine is outlined here. It is based on an Equivalence of Standards in education, training, qualifications, knowledge, skills, competences and the professional conduct associated with specialist practice. In proposing the recognition of specialist practice EFLM has identified 15 EU Member States able to meet Equivalence and in whom the profession and/or its training is regulated (an additional EU Commission requirement). The framework supports and contributes to the Directive's enabling goals for increasing professional mobility, safeguarding consumers and ensuring a more equitable distribution of skills and expertise across the Member States. It represents EFLM's position statement and provides a template for professional societies and/or competent authorities to engage with the EU Commission.

DNA methylation microarrays identify epigenetically regulated lipid related genes in obese patients with hypercholesterolemia.

BACKGROUND: Epigenetics can contribute to lipid disorders in obesity. The DNA methylation pattern can be the cause or consequence of high blood lipids. The aim of the study was to investigate the DNA methylation profile in peripheral leukocytes associated with elevated LDL-cholesterol level in overweight and obese individuals. METHODS: To identify the differentially methylated genes, genome-wide DNA methylation microarray analysis was performed in leukocytes of obese individuals with high LDL-cholesterol (LDL-CH, ≥ 3.4 mmol/L) versus control obese individuals with LDL-CH, < 3.4 mmol/L. Biochemical tests such as serum glucose, total cholesterol, HDL cholesterol, triglycerides, insulin, leptin, adiponectin, FGF19, FGF21, GIP and total plasma fatty acids content have been determined. Oral glucose and lipid tolerance tests were also performed. Human DNA Methylation Microarray (from Agilent Technologies) containing 27,627 probes for CpG islands was used for screening of DNA methylation status in 10 selected samples. Unpaired t-test and Mann-Whitney U-test were used for biochemical and anthropometric parameters statistics. For microarrays analysis, fold of change was calculated comparing hypercholesterolemic vs control group. The q-value threshold was calculated using moderated Student's t-test followed by Benjamini-Hochberg multiple test correction FDR. RESULTS: In this preliminary study we identified 190 lipid related CpG loci differentially methylated in hypercholesterolemic versus control individuals. Analysis of DNA methylation profiles revealed several loci engaged in plasma lipoprotein formation and metabolism, cholesterol efflux and reverse transport, triglycerides degradation and fatty acids transport and ß-oxidation. Hypermethylation of CpG loci located in promoters of genes regulating cholesterol metabolism: PCSK9, LRP1, ABCG1, ANGPTL4, SREBF1 and NR1H2 in hypercholesterolemic patients has been found. Novel epigenetically regulated CpG sites include ABCG4, ANGPTL4, AP2A2, AP2M1, AP2S1, CLTC, FGF19, FGF1R, HDLBP, LIPA, LMF1, LRP5, LSR, NR1H2 and ZDHHC8 genes. CONCLUSIONS: Our results indicate that obese individuals with hypercholesterolemia present specific DNA methylation profile in genes related to lipids transport and metabolism. Detailed knowledge of epigenetic regulation of genes, important for lipid disorders in obesity, underlies the possibility to influence target genes by changing diet and lifestyle, as DNA methylation is reversible and depends on environmental factors. These findings give rise for further studies on factors that targets methylation of revealed genes.

Boronate Affinity Chromatography Accurately Measures HbA1c also in Patients with End-Stage Renal Disease - Performance Evaluation of the A1c HPLC Analyzer.

BACKGROUND: Boronate affinity chromatography is widely used, and the method has lately been improved and designed for HbA1c measurements. We report performance evaluation of the affinity chromatography HbA1c HPLC analyzer. METHODS: Within- and between-run imprecision was assessed based on the results of a series of measurements in three different EDTA blood samples and in control materials. HbA1c levels were measured and compared in 349 EDTA blood samples and 50 samples from patients with end-stage renal disease (ESRD) using the Premier Hb9210 analyzer (affinity chromatography) and the D-10 Hemoglobin Testing System (ion-exchange chromatography). RESULTS: The within- and between-run imprecision CVs ranged from 0.72% to 2.01%. Median HbA1c level measured by the Premier Hb9210 was significantly lower (6.4% [46 mmol/mol] vs. 6.6% [49 mmol/mol], p < 0.001). The Passing-Bablok agreement test yielded a slope of 1.0 (95% CI: 1.0 to 1.0) and intercept of -0.1 (95% CI: -0.1 to -0.1). Correlation coefficient and the mean difference amounted to 0.992 and -0.13% (95% CI: -0.11 to -0.15), respectively. Similar results were obtained for HbA1c levels < 7% [< 53 mmol/mol] and ≥ 7% [≥ 53 mmol/mol]. In ESRD patients, median HbA1c level measured by the Premier Hb9210 was also significantly lower (6.0% [42 mmol/mol] vs. 6.5 [48 mmol/mol], p < 0.001) with the mean difference equal to -0.52% (95% CI: -0.59 to -0.46). CONCLUSIONS: Although the Premier Hb9210 gave lower HbA1c levels, good results agreement with the D-10 Hemoglobin Testing System was found. Analytical performance found for HbA1c measurements in ESRD patients was similar. The Premier Hb9210 analyzer is suitable for routine HbA1c testing in clinical practice.

Diagnostic and prognostic relevance of microparticles in peripheral and uterine blood of patients with endometrial cancer.

OBJECTIVES: Exosomes - microvesicles which are secreted by living cells - can be produced from different cell types and detected in various body fluids. They are the carriers of intercellular information which regulate tumor microenvironment and are considered to be involved in tumor progression and metastasis. Cancer cells can secrete more exosomes than healthy cells, and are expected to be potential tools for tumor diagnosis and treatment. MATERIAL AND METHODS: In this report, we present the results of microparticle analysis in peripheral and uterine blood of patients with endometrial cancer. To the best of our knowledge, this study has been the first to report microvesicle status in peripheral and uterine blood samples. The aim of the study was to determine the amount of total (TF+), endothelial (CD144+) and monocytic (CD14+) microparticles. The counting of the selected microparticles in citrate plasma was performed using flow cytometry on the BD Canto II cytometer. RESULTS: We found that the total amount of microparticles in cancer patients was much higher than in healthy controls. Moreover, microparticle count in uterine blood was higher than in peripheral blood of patients with endometrial cancer. We also demonstrated that the amount of microparticles correlates with the histologic grade and clinical stage of the tumor. CONCLUSIONS: The most interesting finding in this work was the high level of TF, CD144 and CD14 MPs in uterine blood samples. Thus we can consider the monocyte-macrophage-derived MPs as a candidate marker of endometrial cancer and maybe very critical part of the endometrial carcinogenesis.

Carboxylated and undercarboxylated osteocalcin in metabolic complications of human obesity and prediabetes.

BACKGROUND: Carboxylated osteocalcin (Gla-OC) participates in bone remodeling, whereas the undercarboxylated form (Glu-OC) takes part in energy metabolism. This study was undertaken to compare the blood levels of Glu-OC and Gla-OC in nonobese, healthy obese, and prediabetic volunteers and correlate it with the metabolic markers of insulin resistance and early markers of inflammation. METHODS: Nonobese (body mass index [BMI] <30 kg/m2 ; n = 34) and obese subjects (30

The use of selected neutrophil protein plasma concentrations in the diagnosis of Crohn's disease and ulcerative colitis - a preliminary report.

BACKGROUND: Difficulties in diagnosis of inflammatory bowel disease (IBD) motivate the search for new diagnostic tools, including laboratory tests. The aim of this study was to evaluate concentrations of the neutrophil (NEU) proteins leukocyte elastase (HLE-α1AT), lactoferrin and calprotectin as potential biomarkers used in the diagnosis and assessment of clinical activity of Crohn's disease (CD) and ulcerative colitis (UC). MATERIAL/METHODS: The study included 27 patients with CD, 33 patients with UC and 20 healthy controls. Plasma concentrations of calprotectin, lactoferrin and HLE-α1AT were measured using ELISA. RESULTS: In patients with CD higher concentrations of HLE-α1AT (64.3±43.1 vs. 30.1±7.7 ng/l, P<0.001), calprotectin (151.6±97.8 vs. 69.9±22.1 ng/l, P<0.001) and lactoferrin (243.2±102.0 vs. 129.7±32.7 ng/l, P<0.001) than in the control group were found. In patients with UC higher plasma concentrations of HLE-α1AT (62.0±30.9 vs. 30.1±7.7 ng/l, P<0.001), calprotectin (149.6±72.3 vs. 69.9±22.1 ng/l, P<0.001) and lactoferrin (242.6±107.5 vs 129.7±32.7 ng/l, P<0.001) than in the control group were found. HLE-α1AT/NEU and lactoferrin/NEU ratios in patients with UC were significantly higher compared with patients with CD. Calprotectin (P=0.010) and lactoferrin (P=0.023) levels were higher in patients with the active compared with inactive phase of CD. CONCLUSIONS: The diagnostic characteristics of plasma granulocyte protein concentrations indicate the usefulness of these tests in the diagnosis of IBD. Higher HLE-α1AT and lactoferrin/NEU ratios in patients with UC than with CD may suggest the usefulness of these ratios in differential diagnostics. Plasma calprotectin and lactoferrin levels may be useful in CD activity assessment.

Carboxylated and intact osteocalcin predict adiponectin concentration in hemodialyzed patients.

Purpose Disrupted bone metabolism in patients with chronic kidney disease (CKD) is associated with elevated concentrations of biochemical bone markers. Recently, animal studies show the role of osteocalcin in energy metabolism, which is partially confirmed in humans. The aim of our study was to evaluate the relationships between serum concentrations of bone markers and indices of energy metabolism in CKD patients on maintenance hemodialysis; in particular, the relationship between various forms of osteocalcin and adiponectin. Patients and methods The cross-sectional study included 155 hemodialyzed stage 5 CKD patients. Serum concentrations of glucose, insulin, adiponectin, bone alkaline phosphatase (bALP), tartrate resistant acid phosphatase (TRAP), carboxylated (cOC), undercarboxylated (ucOC), and intact osteocalcin (OC) were determined. Results In total cohort, bALP, TRAP, cOC, and ucOC negatively correlated with BMI. All analyzed bone markers positively correlated with adiponectin in total cohort and in men. In multiple linear regression analysis including all patients, log(cOC) and log(intact OC) were the only bone markers that predicted log(adiponectin) (beta = 0.22; p = 0.016 and beta = 0.26; p = 0.010) independently of sex, dialysis vintage, CRP, insulin, iPTH concentrations, BMI, and age. Conclusions Our data confirm the positive association between cOC, intact OC, and adiponectin concentrations in CKD patients on maintenance hemodialysis.

Cannabinoids - a new weapon against cancer?

Cannabis has been cultivated by man since Neolithic times. It was used, among others for fiber and rope production, recreational purposes and as an excellent therapeutic agent. The isolation and characterization of the structure of one of the main active ingredients of cannabis - Δ9 - tetrahydrocannabinol as well the discovery of its cannabinoid binding receptors CB1 and CB2, has been a milestone in the study of the possibilities of the uses of Cannabis sativa and related products in modern medicine. Many scientific studies indicate the potential use of cannabinoids in the fight against cancer. Experiments carried out on cell lines in vitro and on animal models in vivo have shown that phytocannabinoids, endocannabinoids, synthetic cannabinoids and their analogues can lead to inhibition of the growth of many tumor types, exerting cytostatic and cytotoxic neoplastic effect on cells thereby negatively influencing neo-angiogenesis and the ability of cells to metastasize. The main molecular mechanism leading to inhibition of proliferation of cancer cells by cannabinoids is apoptosis. Studies have shown, however, that the process of apoptosis in cells, treated with recannabinoids, is a consequence of induction of endoplasmic reticulum stress and autophagy. On the other hand, in the cellular context and dosage dependence, cannabinoids may enhance the proliferation of tumor cells by suppressing the immune system or by activating mitogenic factors. Leading from this there is a an obvious need to further explore cannabinoid associated molecular pathways making it possible to develop safe therapeutic drug agents for patients in the future.

Relationship between fetuin-A, bone turnover and inflammatory markers concentrations in serum of maintenance hemodialyzed patients.

Introduction: Fetuin-A plays an important role in bone turnover and vascular calcification. Aim: The aim of the study was to assess the relationship between serum fetuin-A concentrations, inflammatory and bone turnover markers of patients on maintenance hemodialysis. Materials and Methods: The study was performed in 71 patients (21 women, 40 men) aged 60 ± 12 years on chronic dialysis because of end-stage renal failure for a period of 75 ± 57.2 months. The routine laboratory tests were performed with Modular P analyzer (Roche Diagnostics), serum concentrations of iPTH were measured using Nichols method, hsCRP and IL-6 using nephelometric techniques while fetuin-A, bone-specific alkaline phosphatase (bALP), fully carboxylated osteocalcin (cOC), undercarboxylated osteocalcin (ucOC), and fibroblast growth factor-23 (FGF-23) were measured using commercially available ELISA kits. Results: Concentrations of fetuin-A were significantly positively correlated with albumin (r=0.37, p=0.003) and negatively associated with patients age (r=26, p=0.04), log (iPTH) (r=0.31, p=0.02), log (CRP) (r=0.31, p=0.02), log (IL-6) (r=0.41, p=0.001), log (ucOC) (r=-0.29, p=0.02), and log (FGF-23) (r=0.27, p=0.04). Conclusions: 1. Patients on maintenance hemodialysis suffer from severe disturbances of bone turnover. 2. Low serum fetuin-A levels are associated with increase markers of bone turnover and inflammation.

Influence of analgesic active 3-[4-(3-trifluoromethyl-phenyl)-piperazin-1-yl]-dihydrofuran-2-one on the antioxidant status, glucose utilization and lipid accumulation in some in vitro and ex vivo assays.

PURPOSE: Earlier we demonstrated that 3-[4-(3-trifluoromethyl-phenyl)-piperazin-1-yl]-dihydrofuran-2-one (LPP1) elevates nociceptive thresholds in the mouse model of diabetic neuropathic pain. Since drug-induced impairments of glucose and lipid metabolism and the oxidative stress might diminish benefits from analgesia achieved by analgesic drugs used in diabetic neuropathy, the effect of LPP1 on glucose utilization, lipid accumulation and its antioxidant and cytotoxic potential were assessed in some in vitro and ex vivo tests. METHODS: Total antioxidant capacity was evaluated spectrophotometrically using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical method, whereas the activities of glutathione (GSH) peroxidase and reductase were measured using methods based on the oxidation of NADPH to NADP. The spectrophotometric method for the evaluation of GSH level in mouse brain tissue homogenates involved the oxidation of GSH by the sulfhydryl reagent 5,5'-dithio-bis(2-nitrobenzoic acid) (DTNB) to form a yellow derivative, 5'-thio-2-nitrobenzoic acid (TNB), measurable at 412 nm. Cytotoxicity and glucose utilization were measured in hepatoma HepG2 cells and in 3T3-L1 adipocytes. Lipid accumulation was measured in 3T3-L1 cell lines. RESULTS: LPP1 had dose-dependent antioxidant properties in DPPH radical assay (14-22% versus control; p < 0.001). Its single administration caused an increase in GSH concentration in brain tissue homogenates of mice by 34% (versus control group; p < 0.05). LPP1 was not cytotoxic and it did not increase glucose utilization or lipid accumulation in cell cultures. CONCLUSIONS: Previously demonstrated antinociceptive properties of LPP1 are accompanied by a lack of cytotoxicity. LPP1 does not impair glucose or lipid metabolism and is an antioxidant. All these properties might be advantageous for its use in diabetic neuropathy.

Autophagy and ubiquitin-dependent proteolysis processes in left ventricular mass loss in pulmonary arterial hypertension.

The goal of this study was to evaluate the intensity of autophagy and ubiquitin-dependent proteolysis processes occurring in myocardium of left ventricle (LV) in subsequent stages of pulmonary arterial hypertension (PAH) to determine mechanisms responsible for LV mass loss in a monocrotaline-induced PAH rat model. LV myocardium samples collected from 32 Wistar rats were analyzed in an early PAH group (n = 8), controls time-paired (n = 8), an end-stage PAH group (n = 8), and their controls (n = 8). Samples were subjected to histological analyses with immunofluorescence staining, autophagy assessment by western blotting, and evaluation of ubiquitin-dependent proteolysis in the LV by immunoprecipitation of ubiquitinated proteins. Echocardiographic, hemodynamic, and heart morphometric parameters were assessed regularly throughout the experiment. Considerable morphological and hemodynamic remodeling of the LV was observed over the course of PAH. The end-stage PAH was associated with significantly impaired LV systolic function and a decrease in LV mass. The LC3B-II expression in the LV was significantly higher in the end-stage PAH group compared to the early PAH group (p = 0.040). The measured LC3B-II/LC3B-I ratios in the end-stage PAH group were significantly elevated compared to the controls (p = 0.039). Immunofluorescence staining showed a significant increase in the abundance of LC3 puncta in the end-stage PAH group compared to the matched controls. There were no statistically significant differences in the levels of expression of all ubiquitinated proteins when comparing both PAH groups and matched controls. Autophagy may be considered as the mechanism behind the LV mass loss at the end stage of PAH.

2024 Guidelines of the Polish Society of Laboratory Diagnostics and the Polish Lipid Association on laboratory diagnostics of lipid metabolism disorders.

Lipid disorders are the most common (even 70%) and worst monitored cardiovascular risk factor (only 1/4 of patients in Poland and in CEE countries are on the low-density lipoprotein cholesterol (LDL-C) goal). To improve this, clear and simple diagnostic criteria should be introduced for all components of the lipid profile. These are the updated guidelines of the two main scientific societies in Poland in the area - the Polish Society of Laboratory Diagnostics (PSLD) and the Polish Lipid Association (PoLA), which, in comparison to those from 2020, introduce few important changes in recommendations (two main lipid targets, new recommendations on LDL-C measurements, calculations new goals for triglycerides, new recommendations on remnants and small dense LDL) that should help the practitioners to be early with the diagnosis of lipid disorders and in the effective monitoring (after therapy initiation), and in the consequence to avoid the first and recurrent cardiovascular events.

Plasma pentraxin 3 concentrations in patients with acute pancreatitis.

BACKGROUND: Pentraxin 3 (PTX3) belongs to the acute phase proteins; its concentration increases significantly in the early stages of inflammation. In nearly 20% of patients with acute pancreatitis (AP) escalated inflammation leads to the development of severe forms of the disease. The aim of this study was to evaluate the pattern of changes in PTX3 concentration in patients with AP at the early stage of the disease (first 5 days from admission) and to assess the relationship between PTX3 and other inflammatory markers. METHODS: The study included 40 patients with AP (16 women and 24 men)--12 with severe and 28 with mild form of AP. Concentrations of PTX3, serum amyloid A (SAA), C-reactive protein (CRP), hepatocyte growth factor (HGF), procalcitonin (PCT), polymorphonuclear elastase (PMN-elastase), interleukin 6 (IL-6), interleukin 18 (IL-18), and soluble receptor for TNFalpha (sTNFR75) were measured in samples collected on the 1st, 3rd, and 5th day of the hospital stay. Plasma PTX3 was measured also in the control group, consisting of 37 age and sex-matched healthy subjects. RESULTS: The highest concentrations of PTX3 were noted on the first day after admission. The concentrations were higher in patients with the severe compared to those with the mild form of AP (median 17.2 vs. 4.0 ng/mL on day 1, p = 0.03; 6.1 vs. 2.2 on day 5, p = 0.044). On each of the study days significant correlations were found between PTX3 and SAA, IL-6, and PMN-elastase (p < 0.05). CONCLUSIONS: The pattern of changes in PTX3 concentration in the early phase of AP is similar to that of IL-6, and its peak levels are achieved earlier compared to CRP. Our findings suggest that PTX3 may be useful in early evaluation and prediction of the severity of AP.

Determination of hepatocyte growth factor at early phase of acute pancreatitis.

AIM: The aim of this study was to assess the diagnostic value of hepatocyte growth factor (HGF) as a new predictor of severity in patients with acute pancreatitis (AP) at early phase of disease. MATERIALS AND METHOD: The studied group involved 40 patients (16 women and 24 men) with AP admitted to Ist Dept. of Surgery Jagiellonian University Medical College in Krakow. Twenty-four patients had mild and twelve severe form of AP. Glasgow and Imrie scores were calculated to evaluate severity of AP. HGF concentrations were measured by ELISA (R&D Systems) on days 1, 3 and 5 after admission within 48 hours after onset of symptoms. RESULTS: Serum median concentrations of HGF was significantly higher in patients with severe versus mild clinical course of AP on each of the study days (7.61 vs 3.30 ng/mL, p = 0.05 on day 1; 7.19 vs 3.43, p = 0.04 on day 3 and 5.76 vs 2.42, p = 0.02 on day 5). HGF positively correlated with Glasgow and Imrie scores (R = 0.57 and R = 0.51). HGF negatively correlated with fetuin A, a negative acute phase protein (R = -0.60 on day 3 and R = -0.45 on day 5) and positively with CRP (R = 0.93; R = 0.80), SAA (R = 0.78; R = 0.82), IL-6 (R = 0.61; R = 0.77; R = 0.85 on day 1, 3 and 5, respectively) and PMN-elastase (R = 0.58; R = 0.64; R = 0.77). On day 1 of the study, HGF reached the diagnostic sensitivity of 100% and specificity of 50% for the detection of severe and moderate AP. CONCLUSIONS: Serum HGF correlates with several inflammatory markers and clinical scores (Glasgow, Imrie) in patients with AP and may be considered a new promising tool in assessing the severity of acute pancreatitis.

[Analysis of selected inflammatory markers for early prediction of severe clinical course of acute pancreatitis]. / Znaczenie wybranych markerów zapalnych dla wczesnego przewidywania ciezkosci przebiegu ostrego zapalenia trzustki.

Despite new diagnostic methods, including novel laboratory parameters and imaging techniques, and growing knowledge on pathogenesis of acute pancreatitis, early assessment of severity remains the main factor influencing prognosis in the disease. The aim of the study was the evaluation of diagnostic accuracy of interleukins (IL): 6 and 18 and acute phase proteins: C-reactive protein (CRP) and serum amyloid A (SAA), together with Glasgow prognostic score during first 48 hours after diagnosing acute pancreatitis in a group of 40 patients treated in the I-st Department of General and Gastrointestinal Surgery University Hospital in Cracow. All the studied inflammatory markers were significantly higher in patients with moderate and severe acute pancreatitis versus patients with mild form of the disease on the first 48 hours of the disease. Expanding Glasgow score with IL-6, IL-18, SAA or CRP determinations resulted in better accuracy for diagnosing severe clinical course of acute pancreatitis.

[Acute pancreatitis and red cell distribution width (RDW)I at early phase of disease]. / Ostre zapalenie trzustki a analiza rozpietosci rozkladu objetosci erytrocytów (RDW) we wczesnej fazie rozwoju choroby.

During last decade, many new biomarkers have been proposed for early diagnosis of acute pancreatitis and prognosis of its severity. However clinical availability of many markers are limited due to costly and time. consuming laboratory methods used, for their assessment, including ELISA technique. Recent studies revealed the usefulness of red cell distribution width (RDW), as a predictor of unfa vorable prognosis in many disease states. RDW is an easily available index generated automatically as a part of standard complete blood count In our group of 40 acute pancreatitis patients, RDW values assessed du ring first 5 days of disease, correlated positively with the duration of hospital stay and the severity of disease as well as with the concentration of selected inflammatory markers. Patients who died had significantly higher RDW comparing to survivors. Our results indicate that RDW may be helpful in early prediction of clinical course of acute pancreatitis.

[The relationship between serum soluble apoptosis marker (sTRAIL) concentration and nutritional parameters in hemodialyzed patients]. / Zaleznosc pomiedzy stezeniem w surowicy rozpuszczalnego markera apoptozy (sTRAIL) a wskaznikami odzywienia u chorych hemodializowanych.

UNLABELLED: TRAIL (TNF-related apoptosis-inducing ligand) acts as a soluble cytokine interacting with transmembrane receptors belonging to the TNF-receptor family. TRAIL can activate both apoptotic and anti-apoptotic signals. Lower levels of serum sTRAIL were inversely associated with a higher risk of all-cause mortality in CKD population. A strong association between malnutrition, inflammation and atherosclerosis, have been observed in CKD patients. The aim of the study was to investigate the relationship between sTRAIL and nutritional markers and adipose tissue metabolism indices in patients on maintenance hemodialysis. The study was performed in group of 76 patients (36 female and 40 male) of average age 60 +/- 12 years on hemodialysis (74.8 +/- 58.0 months). sTRAIL, leptin and adiponectin were determined by ELISA, BMI based on Quetelet formula and serum albumin level using colorimetric bromokrezol green method. The following values of studied parameters were obtained: sTRAIL = 959.6 +/- 204.0 pg/ml, BMI = 24.5 +/- 4.8, leptin = 36.42 +/- 57.94 ng/ml, adiponectin = 17.55 +/- 10.52 microg/ml, leptin/ adiponectin (x10(-3)) = 2.1 +/- 4.5 and albumin = 38.5 +/- 4.5 g/l. sTRAIL correlate negatively with adiponectin and positively with the remaining studied parameters: BMI, albumin, leptin and leptin/adiponectin ratio. CONCLUSION: The observed interrelations between sTRAIL and nutritional parameters as well as studied adipokines may indicate the modulating role of sTRAIL in metabolic regulation.

Analysis of tafazzin and deoxyribonuclease 1 like 1 transcripts and X chromosome sequencing in the evaluation of the effect of mosaicism in the TAZ gene on phenotypes in a family affected by Barth syndrome.

Barth syndrome is a rare disease affecting mitochondria structure and function in males. In our previous study, we have shown a new mutation (c.83T>A, p.Val28Glu) in the TAZ gene in two affected patients with congenital cardiomyopathy. Furthermore, women in this family had no mutations in their blood cells, whereas they only had mutations in the oral epithelial cells. The objective of the project was to evaluate the effect of intertissue mosaicisms on the Barth syndrome phenotypes, searching for another disease-related loci on chromosome X and finally to assess the consequences of the mutation. We conducted the advanced genetic study including cytogenetic research (constitutional karyotyping in blood and fibroblasts), NGS sequencing (with custom chromosome X sequencing together with the evaluation of loss of heterozygosity (LOH) and aberrations (CNV) in the whole genome) in four different tissues and sequencing of tafazzin and deoxyribonuclease 1 like 1 transcripts. The presence of deletions within the 5'untranslated region of the TAZ gene and/or the noncoding regions of the DNASE1L1 gene were detected in several tissues. Whereas, there is no intertissue mosaicism regarding point mutation in TAZ gene in all investigated tissues in female carriers. Only the male patient presented biochemical markers and neurological symptoms of Barth syndrome. All the female carriers are healthy and have normal tafazzin and deoxyribonuclease 1 like 1 transcripts in 2 analyzed tissues. The conclusion of this study is that we cannot rule out or confirm mosaicism in the noncoding regions of TAZ or DNASE1L1 genes, but this is not clinically relevant in female carriers because they are healthy. Finally, it has been proven that mutation (c.83T>A, p.Val28Glu) is responsible for disease in males in this family.