Changes Of Human Serum Albumin Research Articles

Biomolecular interactions of amphotericin B nanomicelles with serum albumins: A combined biophysical and molecular docking approach

In this work, we investigated the interaction of amphotericin B (AmB) nanomicelles on the binding affinity and conformational change of human serum albumin (HSA) in comparison with bovine serum albumin (BSA) under physiological conditions by conducting several spectroscopic techniques further confirmed through molecular docking approaches. The experimental results showed that AmB nanomicelles could bind to both HSA and BSA to form protein/drug complexes with one binding site, and the binding process was spontaneous under physiological conditions. Fluorescence studies revealed that the quenching mechanism of these complexes was static quenching rather than dynamic quenching and exhibited strong binding between serum albumin and AmB nanomicelles. The results from UV–Visible spectra, FT-IR spectra, and CD spectra revealed that the AmB formulations affected the structure of both HSA and BSA proteins by changing the microenvironment around the tryptophan residues of protein and caused a secondary structure change of the protein with the loss of helical stability. The molecular docking experiments also supported the above results and effectively proved the binding and changes in the conformation of serum albumins by AmB micelles. This finding provides information of in vitro drug-plasma protein interactions for further study on the AmB binding mechanism and the pharmacodynamics and pharmacokinetics.

Determination of Amadori Product in Glycated Human Serum Albumin by Spectroscopy Methods

AbstractThe Amadori product is a stable intermediate in the glycation process, and its increased formation is a marker of hyperglycemia in diabetes mellitus. Its accumulation in the body contributes to pathological complications of diabetes including diabetic nephropathy and retinopathy, and cardiovascular diseases. In this study the effect of 3‐β‐hydroxybutyrate on the production of Amadori products and structural changes of human serum albumin (HSA) after incubation with glucose was investigated using FT‐IR, circular dichroism (CD), and UV‐visible and fluorescence spectroscopy. Our results showed that the production of Amadori products in HSA incubated with glucose was decreased in the presence of 3‐β‐hydroxybutyrate. The glycation‐mediated alterations in HSA secondary and tertiary structures were also reduced in the presence of 3‐β‐hydroxybutyrate. These changes were attributed to reduced formation of glucose covalent bonds with the HSA lysine residues in the presence of 3‐β‐hydroxybutyrate. Thus, 3‐β‐hydroxybutyrate reduced the production of Amadori products by reacting with lysine residues and decreasing HSA covalent modifications by glucose.

A comparative study of binding properties of different coumarin-based compounds with human serum albumin

We have disclosed the impact of different coumarin-based compounds (3-(furan-2-yl)-6-methyl-2H-chromen-2-one (FM) and 6-chloro-3-(furan-2-yl)-2H-chromen-2-one) (CM)) on the change of human serum albumin's structure in vitro. The fluorescence experiments demonstrated that the HSA native fluorescence intensity was more reduced by CM than FM followed a static mode. We found the binding constants for FM and CM are 1.289 × 105 M−1 and 6.375 × 105 M−1, respectively, suggesting the much stronger binding affinity of CM to HSA than FM. Thermodynamic analysis showed the hydrophobic interaction between these coumarin-based compounds and HSA were clearly important. Moreover, a series of spectral methods were used to provide more details about the FM/CM-induced conformational changes of HSA. Finally, we used molecular docking set up a new HSA-FM/CM model to predict the possible binding sites.

Size-dependent binding of pristine fullerene (nC60) nanoparticles to bovine/human serum albumin

The size of nC60 nanoparticles may exert profound influence on their binding to serum albumins and their ability to alter protein structure and conformation. To verify this speculation, in this work, five fractions of nC60 dispersions with different particle size distributions were prepared by centrifuge method and characterized using UV–Visible absorption spectroscopy and transmission electronic microscopy. In this work, the binding of five fractions of nC60 nanoparticles with bovine serum albumin (BSA) was demonstrated using fluorescence and synchronous fluorescence spectroscopy. Through the analysis and comparison of binding interactions, it was found that the diameter of nC60 nanoparticles plays a key role in the binding patterns of nC60 nanoparticles to BSA. The smaller nC60 nanoparticles had stronger binding to BSA and exhibited greater influence on the conformation change of BSA. Further study showed that human serum albumin (HSA) has similar binding interaction to nC60 nanoparticles, but the conformation changes of human serum albumin caused by five fractions of nC60 nanoparticles were different from bovine serum albumin. Despite the different particle sizes, the binding sites of nC60 nanoparticles to BSA/HSA were mainly in the vicinity of tryptophan residues. The possible interaction mechanism and the prospects were also discussed.

Morphological changes in human serum albumin in the presence of cationic amphiphilic drugs

Binding of cationic amphiphilic drugs results in unfolding of human serum albumin.

Hyperglycemia induced reactive species trigger structural changes in human serum albumin of type 1 diabetic subjects

Chronic oxidative stress fuels pathogenesis of a large set of diseases. Oxidative stress is the cause and consequence of numerous diseases including type 1 diabetes mellitus (T1DM), in which there is selective destruction of insulin producing pancreatic β-cells. Studies have documented that hyperglycemia produces profound stress. In vivo production of numerous reactive oxygen, nitrogen, chlorine species and lipid/sugar oxidation products in T1DM patients may be the result of persistent hyperglycemia. Post-translational modifications by reactive species may create new antigenic epitopes and play a role in the development of autoimmune response. In this paper our main focus was to establish the effect of existing hyperglycemia induced oxido-nitrosative stress in T1DM patients on the integrity of human serum albumin. Raised nitric oxide, carbonyl, RBC hemolysis, lowered ferric reducing antioxidant power (FRAP), thiol and deformed RBC in T1DM are all highly suggestive of persistent oxido-nitrosative stress. Hyperglycemia induced generation of advanced glycation end products (AGEs) was established by LCMS. Chronic oxido-nitrosative stress can modify HSA in T1DM patients, producing immunologically active albumin. Therefore, it is speculated that the aberrant HSA may play a role in the initiation/progression of T1DM.

Lanthanide Chlorides Decrease the Antioxidant Property of Human Serum Albumin

Substantial conformational changes of human serum albumin, HSA, induced by Lanthanide chlorides (Ce 3+, La 3+, Er 3+ ) in phosphate buffer, 10 mmol L -1 at pH 7.4, was studied by Isothermal Titration Calorimetry, ITC. The strong negative cooperativity of Lanthanide chlorides interaction with HSA recovered from the extended solvation model, indicates that HSA denatures as a result of its interaction with toxic Lanthanide chlorides. DOI: http://dx.doi.org/10.17807/orbital.v9i1.941

Protective effects of curcumin against gamma ray induced conformational change of human serum albumin

This study explores the possibility for protection by curcumin during the molecular and structural changes of human serum albumin (HSA) exposed to gamma irradiation. We used a combination of spectroscopic methods to probe the conformational ensemble of the irradiated HSA and finally evaluated the extent of restoration by curcumin. SDS-PAGE study unfolded the formation of cross linked aggregates as a consequence of increasing exposure of gamma radiation. CD and FTIR spectroscopy indicated significant decrease in the alpha helix content of HSA from 57% to 15% with increasing radiation doses. Steady state and time resolved fluorescence studies complemented the spectroscopic measurements, when lifetime decay was significantly reduced from 6.35ns to 0.37ns. Hydrophobic study showed the effectiveness of curcumin for protection at low dose of gamma irradiated HSA samples. We integrated these observations to investigate protein aggregation under increasing gamma radiation and estimated the same in presence of curcumin. It was elucidated, that when HSA is irradiated at low dose of gamma radiation in presence of curcumin, it is capable of retaining the characteristic properties to a higher extent indicating stabilization of molecular structure of HSA by curcumin. A model for curcumin based protection has been proposed utilizing ThT assay.

Secondary Structural Changes of Intact and Disulfide Bridges-Cleaved Human Serum Albumins in Thermal Denaturation up to 130°C - Additive Effects of Sodium Dodecyl Sulfate on the Changes.

The secondary structural changes of human serum albumin with the intact 17 disulfide bridges (HSA) and the disulfide bridges-cleaved human serum albumin (RCM-HSA) in thermal denaturation were examined. Most of the helical structures of HSA, whose original helicity was 66%, were sharply disrupted between 50 and 100°C. However, 14% helicity remained even at 130°C. The temperature dependence of the degree of disrupted helical structures of HSA was discussed in connection with questions about a general protein denaturation model. When HSA lost the disulfide bridges, about two-thirds of the original helices were disrupted. Although the helices of RCM-HSA remaining after the cleavage of the disulfide bridges were relatively resistant against the heat treatment, the helicity changed from 22% at 25°C to 14% at 130℃. The helicity of RCM-HSA at 130°C agreed with the helicity of HSA at the same temperature, indicating that the same helical moieties of the polypeptides remained unaffected at this high temperature. The additive effects of sodium dodecyl sulfate (SDS) on the structural changes of HSA and RCM-HSA in thermal denaturation were also examined. A slight amount of SDS protected the helical structures of HSA from thermal denaturation below 80°C. Upon cooling to 25°C after heat treatment at temperatures below 70°C with the coexistence of SDS of low concentrations, the helical structures of HSA were reformed to the original level at 25°C before heating. A similar tendency was also observed after heat treatment at 80°C. In contrast, the helical structures of the RCM-HSA complexes with SDS are completely recovered upon cooling to 25°C even after heat treatment up to 100°C. Similar investigations were also carried out on bovine serum albumins which had the intact 17 disulfide bridges and lost all of the bridges.

Quenching of fluorescence by meclizine, a probe study for structural and conformational changes in human serum albumin

The goal of this study was to investigate the interactions between meclizine (MEC) and human serum albumin (HSA) under physiological conditions by different spectroscopies and molecular modeling technique. The drug, MEC quenched the intrinsic fluorescence of HSA and the analysis of the results revealed that static quenching mechanism. The binding of MEC quenches the HSA fluorescence; stoichiometry was 1:1 interaction. Thermodynamic quantities were calculated at different temperatures suggested that hydrophobic and van der Waals interaction with HSA–MEC. The molecular distance, r, between donor and acceptor was estimated according to Forster’s theory of non-radiation energy transfer. CD and FT-IR studies confirm changes of secondary structure of HSA. Molecular docking studies validate MEC molecule interact to HSA in sub domain IIA.

Self-Assembled Nanostructures Based on Activatable Red Fluorescent Dye for Site-Specific Protein Probing and Conformational Transition Detection

Smart and versatile nanostructures have demonstrated their effectiveness for biomolecule analysis and show great potential in digging insights into the structural/functional relationships. Herein, a nanoscale molecular self-assembly was constructed for probing the site-specific recognition and conformational changes of human serum albumin (HSA) with tunable size and emission. A tetraphenylethylene derivative TPE-red-COOH was used as the building block for tailoring fluorescence-silent nanoparticles. The highly specific and sensitive response to HSA was witnessed by the fast turn-on of the red fluorescence and simultaneous disassembly of the nanostructures, whereas various endogenous biomolecules cannot induce such response. The mechanism investigation indicates that the combination of multiple noncovalent interactions is the driving force for disassembling and trapping TPE-red-COOH into HSA. The resultant restriction of intramolecular rotation of TPE-red-COOH in the hydrophobic cavity of HSA induces the significant red emission. By using the fluorescence activatable nanosensor as the structural indicator, the stepwise conformational transitions of HSA during denaturing and the partial refolding of subdomain IIA of HSA were facilely visualized. Benefiting from its activatable signaling, sensitivity, and simplicity, such molecular assembly provides a kind of soft nanomaterial for site-specific biomolecule probing and conformational transition detection concerning their structure, function, and biomedical characteristics.

Binding interaction and conformational changes of human serum albumin with ranitidine studied by spectroscopic and time-resolved fluorescence methods

This study was designed to examine the interaction of histamine H2-receptor antagonist drug ranitidine (RTN) with human serum albumin by multi-spectroscopic methods. The experimental results showed the involvement of dynamic quenching mechanism which was further confirmed by lifetime spectral studies. The binding constants (K a) at three temperatures (288, 298, and 308 K) were 2.058 ± 0.020, 4.160 ± 0.010 and 6.801 ± 0.011 × 104 dm3 mol−1, respectively, and the number of binding sites (m) were 1.169, respectively; thermodynamic parameters ΔH 0 (44.152 ± 0.047 kJ mol−1), ΔG 0 (−26.214 ± 0.040 kJ mol−1), and ΔS 0 (236.130 ± 0.025 J K−1 mol−1) were calculated. The distance r between donor and acceptor was obtained (r = 3.40 nm) according to the Forster theory of non-radiative energy transfer. Synchronous fluorescence, CD, AFM and 3D fluorescence spectral results revealed the changes in secondary structure of the protein upon interaction with RTN. A molecular modeling study further confirmed the binding mode obtained by the experimental studies.

Analysis of Aged Human Serum Albumin Affinity for Doxazosin

Structural changes of human serum albumin (HSA) caused by old age and coexisting diseases result in differences in the binding of doxazosin (DOX). DOX is a postsynaptic α1- adrenoreceptor antagonist used for treatment of hypertension and benign prostatic hyperplasia. In elderly people suffering from various renal or hepatic diseases the significant portion of N-form of human serum albumin (normal) is converted to A-form (aged). The differences in binding of doxazosin to N- and Aform of albumin are an important factor, which may determines therapeutic dosage and toxicity of the test drug. To indicate these differences, the technique of fluorescence spectroscopy was used. The association constant (Ka) obtained from fluorescence quenching demonstrated that doxazosin has higher affinity for AHSA than for HSA. In order to describe the cooperativity in binding process, the values of the Hill's coefficient has been analysed. For DOX-HSA system (λex 295 nm) Hill's coefficient is close to 1 and it indicates that there is a single class of binding sites. For DOX-HSA (λex 275 nm) and DOX-AHSA (λex 275 nm and λex 295 nm) systems we observed positive cooperativity (nH>1). A greater red shift of fluorescence emission maximum of AHSA than HSA in the presence of DOX was observed. This suggests that the binding of DOX to AHSA was accompanied by a stronger increase in polarity around the fluorophores in comparison to HSA. The binding interaction between DOX and HSA has been also studied by molecular docking simulation.

Thermal Fourier Transform Infrared Technique Quickly Detects an Early Onset of Glycation-Induced Conformational Changes of Human Serum Albumin

Numerous potential inhibitors for prevention of advanced glycation end products (AGEs) formation have been extensively investigated for their in vitro and in vivo features, but it would spend a lot of time studying. In the present study, a unique thermal Fourier Transform Infrared (FTIR) combined system as an accelerated method was attempted to simultaneously determine the thermal-dependent conformational changes of human serum albumin (HSA) in the HSA-ribose mixture and examine the onset of the structural transformation from α-helix to β-sheet structures with or without AGEs inhibitors used. The present results clearly indicate that native HSA had an onset temperature at 96°C for the irreversible thermal-induced structural transition from α-helices to β-sheets, whereas HSA-ribose mixture exhibited its onset temperature near at 78°C due to the early occurrence of glycation. However, the onset temperature of the α-helix to β-sheet transition was gradually changed from 78°C to 96°C by increasing the amount of sodium diclofenac or inositol, which was closed to that of the onset temperature of native HSA. This implies that the thermal-induced transition from α-helix to β-sheet for HSA in the HSA-ribose mixture was effectively prevented after adding sodium or inositol. The present study also suggests that this thermal FTIR technique not only rapidly accelerates the conformational changes of HSA-ribose mixture but also directly detects onset temperature of the α-helix to β-sheet transition in real time. This unique thermal FTIR combined system could be a useful tool to screen and evaluate quickly the glycation-induced conformational changes of proteins in a one-step process.

Glycation and secondary conformational changes of human serum albumin: study of the FTIR spectroscopic curve-fitting technique

The aim of this study was attempted to investigate both the glycation kinetics and protein secondary conformational changes of human serum albumin (HSA) after the reaction with ribose. The browning and fluorescence determinations as well as Fourier transform infrared (FTIR) microspectroscopy with a curve-fitting technique were applied. Various concentrations of ribose were incubated over a 12-week period at 37 ± 0.5 ^oC under dark conditions. The results clearly shows that the glycation occurred in HSA-ribose reaction mixtures was markedly increased with the amount of ribose used and incubation time, leading to marked alterations of protein conformation of HSA after FTIR determination.<br /> In addition, the browning intensity of reaction solutions were colored from light to deep brown, as determined by optical observation. The increase in fluorescence intensity from HSA–ribose mixtures seemed to occur more quickly than browning, suggesting that the fluorescence products were produced earlier on in the process than compounds causing browning. Moreover, the predominant α-helical composition of HSA decreased with an increase in ribose concentration and incubation time, whereas total β-structure and random coil composition increased, as determined by curve-fitted FTIR microspectroscopy analysis. We also found that the peak intensity ratios at 1044 cm⁻¹/1542 cm⁻¹markedly decreased prior to 4 weeks of incubation, then almost plateaued, implying that the consumption of ribose in the glycation reaction might have been accelerated over the first 4 weeks of incubation, and gradually decreased. This study first evidences that two unique IR peaks at 1710 cm⁻¹ [carbonyl groups of irreversible products produced by the reaction and deposition of advanced glycation end products (AGEs)] and 1621 cm⁻¹ (aggregated HSA molecules) were clearly observed from the curve-fitted FTIR spectra of HSA-ribose mixtures over the course of incubation time. This study clearly suggests that FTIR spectroscopic curve-fitting technique may be easily used to allow determining the marked changes in the secondary conformational structure and protein aggregation of HSA during ribosylation as well as the production of AGEs.

Underlying the Mechanism of 5-Fluorouracil and Human Serum Albumin Interaction: A Biophysical Study

5-Fluorouracil is clinically utilized as antitumor drug to treat numerous sorts of malignancy, which is made accessible to the objective tissues in conjugation with transport protein serum albumin furthermore which is low harmful when compared to the other drugs of this family and hence its binding characteristics are therefore of prime interest. The steady state and time resolved fluorescence studies, Fourier transform infrared spectroscopy and circular dichroism studies were employed to explain the mode and the mechanism of interaction of 5FU with HSA. 5-Fluorouracil binding is characterized with one high affinity binding site, with the binding constant of the order of 104. The molecular distance r (1.23 nm) between donor (HSA) and acceptor (5-FU) was estimated according to Forster's theory of non-radiative energy transfer. The feature of 5-Fluorouracil induced structural changes of human serum albumin has been studied in detail by Raman spectroscopy, circular dichroism and Fourier transform infrared spectroscopy analysis. The binding dynamics was expounded by synchronous fluorescence spectroscopy, fluorescence lifetime measurements and molecular modelling elicits that hydrophobic interactions and hydrogen bonding, stabilizes the 5-Fluorouracil interaction with HSA.

Hyperglycemia induced structural and functional changes in human serum albumin of diabetic patients: a physico-chemical study.

Structural and functional changes in albumin are of particular interest as numerous studies in vivo have reported a strong involvement of glycated-HSA in the development and progression of chronic diabetic complications. Non-enzymatic addition of glucose molecules to a protein induces structural changes in it. These changes depend on the degree of glycation. In this study, conformational changes in glycated-HSA and its antioxidant capacity were evaluated. HSA was purified from diabetic patients with/without CKD and healthy subjects. Glycation induced an increase in the molecular mass of HSA as determined by mass spectroscopy. Further secondary and tertiary structural changes were observed by UV, circular dichroism (CD) spectroscopy, Fourier transform infrared spectroscopy (FTIR), tryptophan and 1-anilinonaphthalene-8-sulfonic acid (ANS) fluorescence. The mean α-helix content was found to be 59.46% for normal HSA and it was reduced down to 45.63% in HSA isolated from diabetic patients without CKD and to 37.48% in CKD-HSA. FTIR analysis showed Amide I and Amide II band shifting in HSA of diabetic patients without and with CKD. These findings indicate the secondary structure changes in glycated HSA. The tertiary structure is also affected by in vivo glycation as confirmed by intrinsic fluorescence and ANS fluorescence results. Consequently, these structural changes associated with glycation provoked a reduction in the free thiol group and a strong increment of protein carbonyl contents and the fructosamine level in glycated HSA. Antioxidant activity was evaluated by a RBC hemolysis test. The result indicates that the free radical scavenging capacities of HSA were decreased in diabetic patients with or without CKD. Our study revealed that structural and functional features of glycated HSA, isolated from diabetic patients with and without CKD were significantly different from the HSA isolated from non-diabetic subjects. Moreover these changes were more prominent in HSA isolated from diabetic patients with CKD. These findings suggest that active sites of HSA may not be available under extensive glycation, leading to the impairment of its important functions. Thus glycated HSA may be involved in the pathogenesis of diabetes and its complications such as chronic kidney disease.

Ischaemia-modified albumin: a marker of bacterial infection in hospitalized patients with cirrhosis.

Patients with cirrhosis present structural changes of human serum albumin (HSA) affecting non-oncotic functions. Ischaemia-modified albumin (IMA), which reflects the capacity to bind cobalt, has been associated to patient mortality during acute-on-chronic liver failure. This study aimed to assess whether circulating IMA is elevated in advanced cirrhosis and its relationship with severity of cirrhosis and specific complications. A total of 127 cirrhotic patients hospitalized for an acute complication of the disease and 44 healthy controls were enrolled. Plasma IMA and IMA to albumin ratio (IMAr) were measured with a cobalt-binding assay. HSA isoforms carrying post-transcriptional molecular changes were assessed with HPLC-ESI-MS. The effect of endotoxemia on IMA was evaluated in rats with CCl4 -cirrhosis. IMA/IMAr is significantly higher in cirrhotic patients than in controls, but no correlations were found with prognostic scores. IMA did not correlate with the altered HSA isoforms. Ascites, renal impairment and hepatic encephalopathy did not influence IMA/IMAr levels. In contrast, IMA/IMAr is significantly higher in infected than non-infected patients. ROC curves showed that IMA/IMAr had similar discriminating performances for bacterial infection as C-reactive protein (CRP). Moreover, CRP and IMA were independently associated with bacterial infection. Consistently, endotoxin injection significantly increased IMA in cirrhotic, but not in healthy rats. IMA is elevated in patients with advanced cirrhosis. The IMA level does not correlate with disease severity scores, but it is specifically associated to bacterial infection, showing a discriminating performance similar to CRP. Further investigations to assess IMA as a novel diagnostic test for bacterial infection are advocated.

Antioxidant capacity and structural changes of human serum albumin from patients in advanced stages of diabetic nephropathy and the effect of the dialysis.

Changes in the antioxidant capacity of albumin and alterations of the albumin structural conformation were examined in patients in advanced stages of diabetes nephropathy. Human serum albumin was purified from diabetic patients in pre-dialysis (glomerular filtration rate [GFR] between 15 and 29 ml min(-1) 1.73 m(-2)) and those in dialysis (GFR ≤ 15 ml min(-1) 1.73 m(-2)) and then compared with albumin from patients with a normal GFR (>90 ml min(-1) m(-2)). We evaluated the antioxidant capacity of albumin using an enhanced chemiluminescence-based assay and thiol group content, and the structural changes were evaluated by circular dichroism and fluorescence spectroscopy. The antioxidant capacity and thiol content of albumin from patients in advanced stages of diabetic nephropathy were markedly reduced. The circular dichroism spectra showed a mean albumin α-helix content reduction from 44 to 37 % and from 44 to 30 % between the control group and pre-dialysis and dialysis patients, respectively. Additionally, the fluorescence intensity was reduced by 4.2 and 13 % for the groups 4 and 5, respectively, in relation with the control. These data provide evidence for the partial denaturation of albumin and exacerbated oxidative stress among patients in advanced stages of diabetes nephropathy before and even after dialysis.

Conformational and Functional Disturbances of Serum Albumin Are Characteristic for First Episode Drug-naive Schizophrenic Patients.

Objective Conformational changes of human serum albumin (HSA) can disturb its main functions. Aim To reveal serum albumin disturbances using the high-tech approaches in first episode schizophrenia. Methods First-episode drug-naive schizophrenic (FES) patients were investigated. There were employed laser time resolved subnanosecond fluorescence spectroscopy (LTRSFS) using specific albumin fluorescent probe and high resolution nuclear magnetic resonance (NMR) 1 H spectroscopy. Results There were revealed 3 binding sites in albumin molecule with fluorescent decay time of 1, 3 and 9 nanoseconds (A1, A3 and A9 sites, respectively) in healthy volunteers using LTRSFS approach. There was found significant decrease of fluorescent decay time of probe bound to albumin A3 site of FES patients as compared with controls. It points out on the conformational changes in HSA molecule in FES patients. NMR 1 H spectra of blood serum and its albumin fraction of healthy donors and FES patients were studied. There were shown clear differences in NMR 1 H spectra between these two groups that points out to the conformational changes of albumin molecule in FES patients and disturbances in albumin transport functions. The reaction ability of thiol groups of albumin molecules in FES patients was significantly lower in comparison with healthy persons. It also points out on disturbances of antioxidant albumin properties in FES patients. Conclusion High-tech approaches can be useful for the development of new biomarkers for diagnostic and predicting methods and methods for the evaluation of the efficacy of different types of therapy of mental disorders.