Waldemar Balcerzak

Serum and urine metabolomic fingerprinting in diagnostics of inflammatory bowel diseases.

AIM To evaluate the utility of serum and urine metabolomic analysis in diagnosing and monitoring of inflammatory bowel diseases (IBD). METHODS Serum and urine samples were collected from 24 patients with ulcerative colitis (UC), 19 patients with the Crohns disease (CD) and 17 healthy controls. The activity of UC was assessed with the Simple Clinical Colitis Activity Index, while the activity of CD was determined using the Harvey-Bradshaw Index. The analysis of serum and urine samples was performed using proton nuclear magnetic resonance (NMR) spectroscopy. All spectra were exported to Matlab for preprocessing which resulted in two data matrixes for serum and urine. Prior to the chemometric analysis, both data sets were unit variance scaled. The differences in metabolite fingerprints were assessed using partial least-squares-discriminant analysis (PLS-DA). Receiver operating characteristic curves and area under curves were used to evaluate the quality and prediction performance of the obtained PLS-DA models. Metabolites responsible for separation in models were tested using STATISTICA 10 with the Mann-Whitney-Wilcoxon test and the Students t test (α = 0.05). RESULTS The comparison between the group of patients with active IBD and the group with IBD in remission provided good PLS-DA models (P value 0.002 for serum and 0.003 for urine). The metabolites that allowed to distinguish these groups were: N-acetylated compounds and phenylalanine (up-regulated in serum), low-density lipoproteins and very low-density lipoproteins (decreased in serum) as well as glycine (increased in urine) and acetoacetate (decreased in urine). The significant differences in metabolomic profiles were also found between the group of patients with active IBD and healthy control subjects providing the PLS-DA models with a very good separation (P value < 0.001 for serum and 0.003 for urine). The metabolites that were found to be the strongest biomarkers included in this case: leucine, isoleucine, 3-hydroxybutyric acid, N-acetylated compounds, acetoacetate, glycine, phenylalanine and lactate (increased in serum), creatine, dimethyl sulfone, histidine, choline and its derivatives (decreased in serum), as well as citrate, hippurate, trigonelline, taurine, succinate and 2-hydroxyisobutyrate (decreased in urine). No clear separation in PLS-DA models was found between CD and UC patients based on the analysis of serum and urine samples, although one metabolite (formate) in univariate statistical analysis was significantly lower in serum of patients with active CD, and two metabolites (alanine and N-acetylated compounds) were significantly higher in serum of patients with CD when comparing jointly patients in the remission and active phase of the diseases. Contrary to the results obtained from the serum samples, the analysis of urine samples allowed to distinguish patients with IBD in remission from healthy control subjects. The metabolites of importance included in this case up-regulated acetoacetate and down-regulated citrate, hippurate, taurine, succinate, glycine, alanine and formate. CONCLUSION NMR-based metabolomic fingerprinting of serum and urine has the potential to be a useful tool in distinguishing patients with active IBD from those in remission.

Follicular Adenomas Exhibit a Unique Metabolic Profile. 1H NMR Studies of Thyroid Lesions

Thyroid cancer is the most common endocrine malignancy. However, more than 90% of thyroid nodules are benign. It remains unclear whether thyroid carcinoma arises from preexisting benign nodules. Metabolomics can provide valuable and comprehensive information about low molecular weight compounds present in living systems and further our understanding of the biology regulating pathological processes. Herein, we applied 1H NMR-based metabolic profiling to identify the metabolites present in aqueous tissue extracts of healthy thyroid tissue (H), non-neoplastic nodules (NN), follicular adenomas (FA) and malignant thyroid cancer (TC) as an alternative way of investigating cancer lesions. Multivariate statistical methods provided clear discrimination not only between healthy thyroid tissue and pathological thyroid tissue but also between different types of thyroid lesions. Potential biomarkers common to all thyroid lesions were identified, namely, alanine, methionine, acetone, glutamate, glycine, lactate, tyrosine, phenylalanine and hypoxanthine. Metabolic changes in thyroid cancer were mainly related to osmotic regulators (taurine and scyllo- and myo-inositol), citrate, and amino acids supplying the TCA cycle. Thyroid follicular adenomas were found to display metabolic features of benign non-neoplastic nodules and simultaneously displayed a partial metabolic profile associated with malignancy. This finding allows the discrimination of follicular adenomas from benign non-neoplastic nodules and thyroid cancer with similar accuracy. Moreover, the presented data indicate that follicular adenoma could be an individual stage of thyroid cancer development.

Serum and urine 1 H NMR-based metabolomics in the diagnosis of selected thyroid diseases

Early detection of nodular thyroid diseases including thyroid cancer is still primarily based on invasive procedures such as fine-needle aspiration biopsy. Therefore, there is a strong need for development of new diagnostic methods that could provide clinically useful information regarding thyroid nodular lesions in a non-invasive way. In this study we investigated 1H NMR based metabolic profiles of paired urine and blood serum samples, that were obtained from healthy individuals and patients with nodular thyroid diseases. Estimation of predictive potential of metabolites was evaluated using chemometric methods and revealed that both urine and serum carry information sufficient to distinguish between patients with nodular lesions and healthy individuals. Data fusion allowed to further improve prediction quality of the models. However, stratification of tumor types and their differentiation in relation to each other was not possible.

Clinical and histopathological characteristics of patients with incidental and nonincidental thyroid cancer

Introduction Thyroid nodules (TNs) are a common pathology. Their prevalence increases with age. Some of them are suspected of malignancy and qualified for surgery. Sometimes their malignant nature is detected incidentally after a surgical procedure. The aim of the study is to analyze clinical and histopathological characteristics of patients with incidental and nonincidental thyroid carcinoma (ITC vs. NITC). Material and methods The case records of 3,241 patients with solitary and multiple TNs who were treated consecutively between 2008 and 2014 were analyzed retrospectively. After the final selection 235 (7.25%) patients were included in the study (202 females and 33 males, mean age: 52.9 +16.5 years). Seventy-five (31.91%) cases were incidentally diagnosed and 160 (68.09%) were diagnosed before surgery. Results We did not observe any differences in age, gender or stage of disease at the time of diagnosis between the patients with ITC and NITC (p = 0.366, p = 0.850, p = 0.226 respectively). The occurrence of solitary nodules in patients with NITC was significantly higher compared to patients with ITC (p < 0.0001). There were no differences in histopathological types of thyroid cancer (TC). The logistic regression analysis showed that solitary TC was an independent predictor of NITC (p < 0.0001). Conclusions There are no differences in gender, age or histopathological type of cancer in patients with ITC and NITC. Papillary TC is the predominant type in both groups. Incidence of TC in a solitary nodule is significantly higher in NITC than ITC. Solitary type of TC is an independent predictor of NITC. The prevalence of ITC is associated with multifocal type of TC.

Synthesis of fluorescent (benzyloxycarbonylamino)(aryl)methylphosphonates.

Summary The synthesis of a library of structurally variable aromatic esters of (benzyloxycarbonylamino)(aryl)methylphosphonic acids is described by means of the Oleksyszyn reaction. The library was enlarged by the application of a Suzuki–Miayra approach and by preparation of mixed esters.

Diagnostics of Thyroid Malignancy and Indications for Surgery in the Elderly and Younger Counterparts: Comparison of 3,749 Patients

Background It seems valuable for clinicians to know if diagnostics of thyroid malignancy (TM) and indications for surgery in the elderly patients differ from these in younger counterparts. Materials and Methods Retrospective analysis of the medical records of 3,749 patients surgically treated for thyroid tumor. Data of patients with histopathology confirmed TM (n = 309) were studied. Results The rate of cytological prediction to malignancy was more than three times higher in elderly women. Compression was a main reason for surgery in the elderly (p < 0.0001). The final diagnosis of malignancy was significantly higher in older women (p = 0.002). Clinical suspicion of malignancy was positively correlated with histopathological diagnosis in total group of women (r = 0.543, p < 0.001) and total group of men (r = 0.560, p < 0.001). The subgroup of the eldest TM patients included a significantly higher number of subjects with advanced cancer and primary tumor progression (p < 0.0001). Distant metastases were significantly more presented among the elderly patients (p = 0.032). Conclusions The rate of cytological prediction to malignancy in elderly women is high. Tracheal compression is a common surgical indication in the elderly patients. The final diagnoses of malignancy predominate in elderly women. The oldest TM patients present a higher number of advanced thyroid tumors and distant metastases.

A useful procedure for detection of polyamines in biological samples as a potential diagnostic tool in cancer diagnosis

BackgroundPolyamines present in human body are frequently considered as markers of occurrence of cancer. Therefore, the availability of simple and efficient method for determination of their level in body liquids and tissues is of some interest.MethodsSupported liquid membrane technology coupled with HPLC seems to be an appropriate technique to follow the level of polyamines in human blood and urine. Thus, the membranes of two different geometries: flat sheet and hollow fiber were studied as a mean for separation and enrichment of studied polyamines from urine and tissue samples in order to prepare samples to be analyzed by HPLC.ConclusionsDeveloped extraction systems offer an interesting alternative to traditional techniques such as: liquid-liquid or solid-phase extraction due to several features, which are: very high enrichment of polyamines without previous work-up, simple procedure of extraction and tiny volume of organic solvents used. This enables efficient determination of their levels in body liquids.