Adriana Mika

Current methods of the analysis of immunosuppressive agents in clinical materials: A review

More than 100000 solid organ transplantations are performed every year worldwide. Calcineurin (cyclosporine A, tacrolimus), serine/threonine kinase (sirolimus, everolimus) and inosine monophosphate dehydrogenase inhibitor (mycophenolate mofetil), are the most common drugs used as immunosuppressive agents after solid organ transplantation. Immunosuppressive therapy, although necessary after transplantation, is associated with many adverse consequences, including the formation of secondary metabolites of drugs and the induction of their side effects. Calcineurin inhibitors are associated with nephrotoxicity, cardiotoxicity and neurotoxicity; moreover, they increase the risk of many diseases after transplantation. The review presents a study of the movement of drugs in the body, including the processes of absorption, distribution, localisation in tissues, biotransformation and excretion, and also their accompanying side effects. Therefore, there is a necessity to monitor immunosuppressants, especially because these drugs are characterised by narrow therapeutic ranges. Their incorrect concentrations in a patients blood could result in transplant rejection or in the accumulation of toxic effects. Immunosuppressive pharmaceuticals are macrolide lactones, peptides, and high molecular weight molecules that can be metabolised to several metabolites. Therefore the two main analytical methods used for their determination are high performance liquid chromatography with various detection methods and immunoassay methods. Despite the rapid development of new analytical methods of analysing immunosuppressive agents, the application of the latest generation of detectors and increasing sensitivity of such methods, there is still a great demand for the development of highly selective, sensitive, specific, rapid and relatively simple methods of immunosuppressive drugs analysis.

The relationship between specific Fatty acids of serum lipids and serum high sensitivity C- reactive protein levels in morbidly obese women.

Background/Aims: The fatty acid profile in plasma lipids contributes to the increase of plasma high sensitivity C-reactive protein (hsCRP), a marker of inflammation and predictor of cardiovascular risk. The aim of this study was to examine the relationship between specific fatty acids (FA) of serum lipids and serum hsCRP in morbidly obese woman. Methods: The study included 16 morbidly obese (mean BMI= 43 ± 2.2 kg/m2) non-diabetic woman awaiting bariatric surgery. FA extracted from serum lipids were methylated and analyzed on GC-MS. Commercially available ELISA kits were used to determine the serum inflammatory markers. Results: We demonstrated that total saturated FA (SFA) and total monounsaturated FA (MUFA) of serum lipids were positively correlated with serum hsCRP, whereas both n-3 and n-6 total polyunsaturated FA (PUFA) were negatively correlated with serum hsCRP. Serum interleukin-6 correlated positively with some SFA and MUFA, whereas negatively with some of PUFA. Positive correlation between serum hsCRP and specific SFA and MUFA or negative correlation with PUFA decreased with the increased FA chain length. The number and localization of double bonds also had impact on these correlations. Conclusion: Our findings suggest that individual serum lipid FA levels, depending on the length of FA chain, number and the localization of double bonds are distinctly associated with hsCRP in morbidly obese subjects.

A comprehensive study of serum odd- and branched-chain fatty acids in patients with excess weight.

While small amounts of odd‐chain fatty acids (OCFAs) and branched‐chain fatty acids (BCFAs) were known to be present in mammals, it was quite recently that they were shown to play an important role in human health. However, still little is known on OCFA and BCFA profiles in subjects who have obesity. The aim of this study was to verify whether obesity is associated with changes in serum OCFA and BCFA profiles.

Alterations of specific lipid groups in serum of obese humans: a review.

Obesity is a major contributor to the dysfunction of liver, cardiac, pulmonary, endocrine and reproductive system, as well as a component of metabolic syndrome. Although development of obesity‐related disorders is associated with lipid abnormalities, most previous studies dealing with the problem in question were limited to routinely determined parameters, such as serum concentrations of triacylglycerols, total cholesterol, low‐density and high‐density lipoprotein cholesterol. Many authors postulated to extend the scope of analysed lipid compounds and to study obesity‐related alterations in other, previously non‐examined groups of lipids. Comprehensive quantitative, structural and functional analysis of specific lipid groups may result in identification of new obesity‐related alterations. The review summarizes available evidence of obesity‐related alterations in various groups of lipids and their impact on health status of obese subjects. Further, the role of diet and endogenous lipid synthesis in the development of serum lipid alterations is discussed, along with potential application of various lipid compounds as risk markers for obesity‐related comorbidities.

Identification of lipids in the cuticle of the parasitic nematode Anisakis simplex and the somatic tissues of the Atlantic cod Gadus morhua

The main aim of this work was to assign the cuticular lipids identified in a parasitic nematode and to distinguish those originating from its host. The hypothesis that long-chained fatty acids and sterols are imported by the parasite in the absence of certain enzymes was also tested. The organisms (Anisakis simplex and Gadus morhua) were extracted in petroleum ether and dichloromethane. Matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF) was used to identify unknown components, and electrospray ionization mass spectrometry (ESI/MS) to verify recognized groups of lipids. The lipid classes identified in the surface layer were free saturated and unsaturated fatty acids, triacylglycerols, sterols and non-polar sphingolipids (ceramides, sphingoid bases). The most abundant fraction consisted of fatty acids. The predominant saturated acids were tetradecanoic acid in the petroleum ether extract of A. simplex, hexadecanoic acid in the dichloromethane extract of A. simplex, and also the polyunsaturated octadecahexaenoic and octadecatrienoic acids in both extracts of the parasitic nematode. The mass spectrum revealed the presence of fatty acids with different numbers of carbons, and with odd and even numbers of unsaturated bonds. The MALDI-TOF mass spectrum also identified triacylglycerols (TAGs). The dominant short-chain TAGs were CoCoCy:(1), CoCoPg and Bu0:0B:(6). The majority of TAGs were found in the ether and dichloromethane extracts of A. simplex. Sterols were the least common class of lipids found in the nematode extracts; most likely, this is the fraction that is entirely incorporated from the host organism because of the parasites inability to synthesize them. MALDI-TOF also identified non-polar sphingolipids--ceramides and sphingoid bases. The signals due to N-octanoyl-D-erythro-octasphinganine (m/z 288.3) and N-tetranoyl-D-erythro-tetradecasphinganine (m/z 316.4) were dominant on the mass spectra; quite a large number of short-chain non-polar sphingolipids were also identified.

Increased Serum Level of Cyclopropaneoctanoic Acid 2-Hexyl in Patients with Hypertriglyceridemia-Related Disorders

We recently reported the presence of various cyclopropane fatty acids—among them, cyclopropaneoctanoic acid 2-hexyl—in the adipose tissue of obese women. The aim of this study was to verify whether the presence of cyclopropaneoctanoic acid 2-hexyl in human serum was associated with obesity or chronic kidney disease (both being related to dyslipidemia), and to find potential associations between the serum level of this compound and specific markers of the these conditions. The serum concentration of cyclopropaneoctanoic acid 2-hexyl was determined by gas chromatography–mass spectrometry (GC–MS) in non-obese controls, obese patients, obese patients after a 3-month low-calorie diet, and individuals with chronic kidney disease. Obese patients and those with chronic kidney disease presented with higher serum levels of cyclopropaneoctanoic acid 2-hexyl than controls. Switching obese individuals to a low-calorie (low-lipid) diet resulted in a reduction in this fatty acid concentration to the level observed in controls. Cyclopropaneoctanoic acid 2-hexyl was also found in foods derived from animal fat. Serum concentrations of triacylglycerols in the analyzed groups followed a pattern similar to that for serum cyclopropaneoctanoic acid 2-hexyl, and these variables were positively correlated with each other among the studied groups. Patients with hypertriglyceridemia-related conditions presented with elevated serum levels of cyclopropaneoctanoic acid 2-hexyl. Our findings suggest that its high serum level is related to high serum triacylglycerol concentrations rather than to body mass or BMI.

Disorders of serum omega-3 fatty acid composition in dialyzed patients, and their associations with fat mass

Abstract Patients with chronic kidney disease (CKD) are at increased risk of cardiovascular mortality. Lipid disorders, a constant feature of CKD, might contribute to this state. The aim of this study was to evaluate n-3 polyunsaturated fatty acids (PUFA) composition in CKD patients treated with dialysis, in comparison to the general population and to assess possible associations between the n-3 PUFA profile and anthropometric variables. Thirty-three prevalent dialysis patients were studied and compared with an age- and sex-adjusted control group of 22 patients. Fatty acid composition in serum was analyzed by gas chromatography with a mass spectrometer detector (GC-MS) and anthropometric measures were assessed by bioimpedance spectroscopy. The fatty acid profile of dialyzed patients was characterized by a significantly lower percentage content of n-3 PUFA. For α-linolenic acid (ALA), it was 0.21 ± 0.09% in dialysis patients versus 0.33 ± 0.11% in the control group (p < .001). For eicosapentanoic acid (EPA), 0.59 ± 0.23% versus 1.15 ± 0.87% (p < .001), and for docosahexaenoic acid (DHA) 1.11 ± 0.50% versus 1.75 ± 0.87% (p < .001), respectively. The amount of n-3 PUFA decreased with time on dialysis and it correlated positively with body fat mass. For DHA, this correlation was r = .48 (p < .01) and for EPA r = .40 (p < .05). Patients with CKD have a relatively low content of n-3 PUFA which may contribute to their high cardiovascular risk. Patients with a higher content of body fat are characterized by a favorable fatty acid composition.

Composition of fatty acids and sterols composition in brown shrimp Crangon crangon and herring Clupea harengus membras from the Baltic Sea

The composition and the content of lipids, fatty acids and sterols during spawning (spring) in different tissues of herring Clupea harengus membras were compared with quantities of lipid compounds in abdomen muscle of brown shrimp Crangon crangon. The largest quantity of fatty acids in lipids (93.7%), was observed in fish muscle. The liver was characterized by a higher content and variety of sterols (about 4.6 times more than in fish muscle), and in fish sperm, additional fatty acids were identified and the highest amount of EPA and DHA (22% and 34% of fatty acids, respectively) was recorded. The brown shrimp, despite its small size, contained significantly more lipids than the Baltic herring per g of tissues. Fatty acids were at the same level (83% of the total lipids in shrimp muscle and 93.7% in fish muscle), but the amount of sterols was significantly higher in the muscle of shrimp (5.50 ±0.31 mg g−1, 17% of total sterols, n=10) than in fish muscle (1.33 ±0.04 mg g−1, 6.3% of total lipids, n=6). And thus, shrimp is a good source of food for higher trophic levels, and in consequence — a good source of PUFAs for humans. With these results we prove that shrimp and herring play an important role in the supply of EFAs, which has great pharmaceutical and medical benefits.

Application of nuclear magnetic resonance spectroscopy for the detection of metabolic disorders in patients with moderate kidney insufficiency

Graphical abstract Figure. No caption available. HighlightsUsing 1H NMR spectroscopy and multivariate analysis we found significant differences in metabolite profiles of patients and healthy controls.Our study provided a valuable insight into interactions between renal function and metabolite profile of human serum.MetPA analysis demonstrated significant intergroup differences in 5 potential target pathways and 14 metabolites. ABSTRACT Chronic kidney disease (CKD) is one of the major problems of modern medicine and a huge socioeconomic burden. Thorough knowledge of metabolic alterations associated with this condition is vital to prevent its progression. However, still little is known about metabolic disorders associated with CKD. In this study, we used 1H NMR spectroscopy and multivariate analysis to identify alterations in serum metabolites of patients with various stages of CKD. 1H NMR spectroscopy followed by multivariate analysis showed that CKD patients differed from the controls in terms of 15 endogenous metabolites, and MetPA analysis demonstrated significant intergroup differences in 5 potential target pathways and 14 metabolites. Owing a good performance of discriminant models, these findings suggest that CKD patients and healthy controls differ in terms of their metabolic fingerprints. In turn, the results of MetPA analysis imply that CKD and its progression exert an effect on selected metabolic pathways. This study provided a better insight into metabolic alterations associated with CKD, and identified some target pathways that can be potentially modified to slow down the progression of this serious and debilitating disease.

Hyper-Elongation in Colorectal Cancer Tissue – Cerotic Acid is a Potential Novel Serum Metabolic Marker of Colorectal Malignancies

Backgrounds/Aims: Colorectal cancer (CRC) cells show some alterations of lipid metabolism. Elongation of fatty acids (FA) has not been studied in CRC tissues thus far. The aim of this study was to verify if CRC specimens and normal colon mucosa differ in terms of their levels of very long-chain FAs, a product of FA elongation. Moreover, the expression of elongase genes has been studied in normal tissue and CRC. Finally, we searched for some specific products of FA elongation in serum of CRC patients. Methods: The specimens of normal colon mucosa and CRC were obtained from nineteen CRC patients differ in terms of FA elongation. We also searched for some specific products of FA elongation in serum of CRC patients and from healthy volunteers. Tissue and serum FA profiles were determined by means of gas chromatography-mass spectrometry (GC/MS), and the tissue expression of elongases (ELOVLs) was analyzed with real-time PCR. Results: Compared to normal colon tissue, CRC specimens showed significantly higher levels of 22-, 24- and 26-carbon FAs, stronger expressions of ELOVL1 and ELOVL6 (4- and 9-fold elevated respectively), and higher values of 18: 0/16: 0 elongation index. We also demonstrated presence of cerotic acid (26: 0) in serum of all CRC patients but in none of the healthy controls. Conclusions: CRC tissue seems to be characterized by enhanced FA elongation (hyper-elongation). Presence of cerotic acid in CRC patients sera and absence of this FA in healthy subjects points to this compound as a strong candidate for specific metabolic marker of colorectal malignancies.