Biological Fluids Of Patients Research Articles

3-Hydroxy-3-Methylglutaric Acid Disrupts Brain Bioenergetics, Redox Homeostasis, and Mitochondrial Dynamics and Affects Neurodevelopment in Neonatal Wistar Rats.

3-Hydroxy-3-methylglutaric acidemia (HMGA) is a neurometabolic inherited disorder characterized by the predominant accumulation of 3-hydroxy-3-methylglutaric acid (HMG) in the brain and biological fluids of patients. Symptoms often appear in the first year of life and include mainly neurological manifestations. The neuropathophysiology is not fully elucidated, so we investigated the effects of intracerebroventricular administration of HMG on redox and bioenergetic homeostasis in the cerebral cortex and striatum of neonatal rats. Neurodevelopment parameters were also evaluated. HMG decreased the activity of glutathione reductase (GR) and increased catalase (CAT) in the cerebral cortex. In the striatum, HMG reduced the activities of superoxide dismutase, glutathione peroxidase, CAT, GR, glutathione S-transferase, and glucose-6-phosphate dehydrogenase. Regarding bioenergetics, HMG decreased the activities of succinate dehydrogenase and respiratory chain complexes II-III and IV in the cortex. HMG also decreased the activities of citrate synthase and succinate dehydrogenase, as well as complex IV in the striatum. HMG further increased DRP1 levels in the cortex, indicating mitochondrial fission. Finally, we found that the HMG-injected animals showed impaired performance in all sensorimotor tests examined. Our findings provide evidence that HMG causes oxidative stress, bioenergetic dysfunction, and neurodevelopmental changes in neonatal rats, which may explain the neuropathophysiology of HMGA.

Development of an immunochromatographic test strip to detect pneumolysin by monoclonal antibody capture method

BackgroundThe currently available methods for the detection of Streptococcus pneumoniae are labour-intensive, and involve time-consuming steps needing skilled technicians to perform the assays. Therefore, we developed a rapid lateral flow based immunochromatographic test strip for detection of S. pneumoniae using pneumolysin using monoclonal antibody capture approach. Methodology − The strip was designed by assembling a sample pad, conjugation pad, nitrocellulose membrane and adsorbent pad supported by a plastic backing card. Optimization of control spot was done by using three different concentrations of monoclonal antibody (0.1 µg, 0.25 g and 0.5 µg). Monoclonal antibody coupled with gold nanoparticles was immobilized on conjugate pad. Recombinant pneumolysin at three different concentrations (10 ng/ml, 50 ng/ml and 100 ng/ml) was used. Diluted pneumolysin was poured onto the sample pad which migrated through the conjugate pad containing gold antibody conjugate and the antigen–antibody complex migrated to the test spot to bind to the polyclonal antibody. The remaining free antibody bound to the control spot to visualize a control spot. Results: Control spot was optimized at 0.5 µg of mouse IgG. Of three different concentrations of Pneumolysin, the spot was best visualized at 50 ng/ml Pneumolysin concentration. Conclusion: Our ICT based device could detect a commercially available recombinant Pneumolysin of S. pneumoniae. Future study is directed to validate the strip for detection of Pneumolysin using biological fluids of patients (particularly children) infected with S. pneumoniae.

Chemical and physical features of biological fluids in treatment of hydatid disease.

The aim of this paper is to better understand the dynamics of crystallogenic and starting activity in biological fluids of patients throughout surgery and the late postoperative phase in alveococcosis. Samples of saliva from 22 individuals with alveococcosis were included in the research. Biological fluid samples were taken at the time of admission and before the patient was discharged. Following that, slides were made utilizing the teziocrystalloscopy method, which incorporates the investigation of the crystal forming activity of mixed saliva with its starting characteristics using a 0.9 percent sodium chloride solution as the foundation ingredient. Using our own set of criteria, we evaluated the outcomes of crystalloscopic and tezigraphic experiments. Specrophotometric examination of tezigraphic and crystalloscopic facies was done using a PowerWave XS microplate spectrophotometer at wavelengths of 400, 350, and 300 nm to augment the results from ocular morphometry of dried saliva micro slides. Surgical therapy results in a partial normalization of physical and chemical parameters, as well as the composition of the patient's biological fluids after the patient is discharged from the hospital.

Meta-Analysis of Assessment of Total Oxidative Stress and Total Antioxidant Capacity in Patients with Periodontitis.

Periodontitis is intricately linked to oxidative stress-antioxidant (redox) imbalance. The antioxidant system scavenges the oxygen free radicals in biological fluids in patients with periodontitis. However, little is still known about the free radicals mediated oxidative stress and reductive ability of the antioxidant system. Thus, the present meta-analysis aims to quantitatively review the literature that assessed the oxidative stress marker total oxidative stress (TOS) and total antioxidant capacity (TAC) in various biological fluids of patients with periodontitis. Methodology. Electronic databases were searched for studies that assessed TOS and TAC levels in various biological samples of patients with periodontitis. From the 1,812 articles identified, 1,754 were excluded based on title and abstract screening due to irrelevance to the topic of interest. A full-text assessment of the remaining 58 articles led to the selection of 42 articles that satisfied the inclusion criteria. Of these, only 24 studies had consistent data for quantitative analysis. The periodontitis group displayed significantly elevated TOS levels (p < 0.05) in serum, gingival crevicular fluid (GCF), and saliva samples in the studies evaluated. In contrast, the periodontitis group exhibited significantly attenuated TAC levels (p < 0.01) compared to healthy controls in plasma, serum, and GCF samples of the studies evaluated, which was insignificant in salivary samples (p=0.433). At the same time, the periodontitis group displayed insignificantly elevated TAC levels after periodontal therapy (p=0.130). The present meta-analysis showed significantly higher TOS and lower TAC in periodontitis, reflecting the elevated oxidative stress level than the control group. Clinical Relevance. Scientific rationale for the study: The imbalance between oxidants and antioxidants (oxidative stress (OS)) plays a critical role in the onset and progression of periodontitis; the assessment of the relationship between OS-related biomarkers in regional samples and systemic samples of patients with periodontitis helps us to evaluate the periodontal disease progression. The OS biomarker levels can be used to assess periodontal disease and therapeutic efficacy.

The effectiveness of medical personnel vaccination against hepatitis B

Medical personnel belong to the occupational risk group of hepatitis B virus (HBV) infection due to the increase of parenteral interventions and procedures using sophisticated equipment, constant contact with biological fluids of patients, the widespread use of antibiotics and cytostatics that have an immunosuppressive effect, etc. Insufficient objectivity in assessing the quality of hepatitis B (HBV) immunoprophylaxis only according to documentation may be associated with a violation of immunization regimens, the presence of chronic pathology in vaccinated, age-related immunodeficiency. Despite the effectiveness of HCV vaccination, the protective activity of specific humoral immunity (antibodies to HBsAg — anti-HBs) among vaccinated medical workers has not been sufficiently studied. The results of studying the intensity and duration of the immune response in 2075 employees of two large multidisciplinary hospitals vaccinated against HBV (average age 46.3 ± 10.1 years) are presented. A high level of vaccination coverage of medical personnel in hospitals No. 1 and No. 2 was established, amounting to 96.6 % and 97.2 %, and the level of collective immunity was equal to 1645.5±72.5 IU/l and 886±123.7 IU/l, respectively. The absence of cases of HBV in vaccinated individuals during immunoprophylaxis according to the standard scheme was noted. It seems advisable to use an individual approach to the tactics of revaccination when reducing the content of anti-HBs in the blood to low concentrations (10–100 IU / l), preventing them from falling below the protective level.

A study of connective tissue metabolites in biological fluids of patients with various types of glaucoma

PURPOSE. To quantify the content of connective tissue metabolites reflecting the exchange of its main components in biological fluids in low tension glaucoma (LTG) and primary open-angle glaucoma (POAG), and to conduct a comparative analysis of the obtained data. METHODS. The studies were conducted in 33 patients with low tension glaucoma aged 51 to 80 years old (mean age 69 ± 10.4 years), 20 patients with POAG aged 53 to 80 years old (mean age 67 ± 10.1 years) and 15 clinically healthy volunteers aged 52 to 78 years old (mean age 68 ± 9.2 years). The protein components of the fibrous part of the connective tissue were analyzed for daily excretion of oxyproline in urine by the method involving para-Dimethylamin-obenzaldehyde modified by P.N. Sharaev. Serum glycosaminoglycans (GAG) were studied by analyzing carbohydrate components of connective tissue. The concentration of GAG in the blood serum was determined by the colorimetric method, using Dische's carbazole reaction. RESULTS. The amount of GAG in the blood serum of LTG patients was 3.26 ± 1.57 mmol/L, while in the control group consisting of clinically healthy volunteers — 2.92 ± 1.25 mmol/L. The differences were not statistically significant (p &gt; 0.1). The amount of daily excretion of oxyproline in the urine of LTG patients was 17.17 ± 8.64 mmol/day, and in the group of clinically healthy individuals — 13.96 ± 5.61 mmol/day. The differences were statistically significant (p &lt; 0.05). The amount of GAG in the blood serum of POAG patients was 3.97 ± 1.17 mmol/L. The differences in relation to LTG were statistically significant (p &lt; 0.05). The amount of daily excretion of oxyproline in the urine of POAG patients was 32.71 ± 19.79 mmol/day. The differences were statistically significant in relation to LTG (p &lt; 0.05). The amount of GAG and oxyproline in biological fluids tends to increase with the advancement of LTG and POAG. CONCLUSION. Destruction processes in the connective tissue do occur in LTG, but are less pronounced in comparison to POAG and more pronounced in comparison to healthy individuals. The indicator that most clearly reflects the activity of destructive processes in the connective tissue is the daily excretion of oxyproline in urine.

Therapeutic exosomes for various human diseases: Special issue of BMB Reports in 2022

Extracellular vesicles (EVs), especially exosomes, are cell-derived nanoparticles harboring various cellular components such as RNAs, lipids, and proteins for intercellular communication. Roles of EVs as intercellular communicators have been extensively studied in the last few decades, especially under various pathological conditions. Deciphering the message in EVs isolated from biological fluids of patients can provide valuable information not only for disease diagnosis, but also for disease monitoring or treatment responses. EVs are also attractive treatment modality and drug delivery system with favorable properties of biocompatibility, selective tropism, and stability. Stem cell-derived naïve EVs have been tested for their regenerative or immunomodulatory effects in numerous preclinical and clinical studies. This so-called "cell-free cell therapy" is supported by the idea that most therapeutic actions of conventional cell therapy are mediated by paracrine action of EVs released from stem cells. In that sense, immune cell-derived EVs are regarded as a reasonable option for cancer immunotherapy. Such therapeutic effect of EVs can be dramatically augmented by incorporating active pharmaceutical ingredients (APIs) to make engineered exosomes as "Trojan Horses". Biomimetic EVs or cell-derived nanovesicles can be generated through various physicochemical methos such as serial extrusion. They provide alternative options due to their high productivity and relatively easy purification. In this special issue, therapeutic applications of naïve or engineered EVs are discussed in various human diseases including cardiovascular diseases, renal disorders, neurological diseases, cancers, and infectious diseases focusing on COVID-19. [BMB Reports 2022; 55(1): 1-2].

S-adenosylmethionine induces mitochondrial dysfunction, permeability transition pore opening and redox imbalance in subcellular preparations of rat liver.

S-adenosylmethionine (AdoMet) predominantly accumulates in tissues and biological fluids of patients affected by liver dysmethylating diseases, particularly glycine N-methyltransferase, S-adenosylhomocysteine hydrolase and adenosine kinase deficiencies, as well as in some hepatic mtDNA depletion syndromes, whose pathogenesis of liver dysfunction is still poorly established. Therefore, in the present work, we investigated the effects of S-adenosylmethionine (AdoMet) on mitochondrial functions and redox homeostasis in rat liver. AdoMet decreased mitochondrial membrane potential and Ca2+ retention capacity, and these effects were fully prevented by cyclosporin A and ADP, indicating mitochondrial permeability transition (mPT) induction. It was also verified that the thiol-alkylating agent NEM prevented AdoMet-induced ΔΨm dissipation, implying a role for thiol oxidation in the mPT pore opening. AdoMet also increased ROS production and provoked protein and lipid oxidation. Furthermore, AdoMet reduced GSH levels and the activities of aconitase and α-ketoglutarate dehydrogenase. Free radical scavengers attenuated AdoMet effects on lipid peroxidation and GSH levels, supporting a role of ROS in these effects. It is therefore presumed that disturbance of mitochondrial functions associated with mPT and redox unbalance may represent relevant pathomechanisms of liver damage provoked by AdoMet in disorders in which this metabolite accumulates.

Abstract P-43: Effect of Glucocerebrosidase Dysfunction on the Pool of Plasma Exosomes of Patients with Gaucher Disease

Background: Extracellular vesicles (EVs) are small membrane vesicles released from different types of cells. EVs are found in many human biological fluids. Exosomes are a subtype of EVs that are released by the fusion of multivesicular bodies with the plasma membrane. This type of vesicles is characterized by specific exosomal markers. Exosomes extracted from peripheral body liquids could have specific properties associated with different physiological conditions as well as human disorders, including neurodegenerative diseases. Gaucher disease (GD) – is the most common form of lysosomal storage disorders caused by mutations in the glucocerebrosidase (GBA) gene. Lysosome functionality is critical for the regulation of extracellular vesicle secretion and content. In model animals, the inhibition of glucocerebrosidase has been shown to increase the secretion of extracellular vesicles in brain tissues. Amount evaluation of EVs and their size in the biological fluids of patients with GD has not been early performed; therefore, it is unknown whether lysosomal dysfunction found in GD patients influences the plasma pool of EVs. The aim of this study was to evaluate the amount of blood plasma EVs in patients with GD and their characterization for morphology and size. Methods: EVs were isolated from the blood plasma of 8 GD patients and 8 controls by ultracentrifugation, and were characterized using cryo-electron microscopy (cryo-EM), nanoparticle tracking analysis (NTA), and dynamic light scattering (DLS). Also, the presence of exosomal markers CD9, CD63, CD81, and HSP70 was analyzed by flow cytometry and western blot. Results: Here, it was first shown an increased proportion of exosome fraction in EVs from plasma of GD patients compared to controls by DLS and cryo-EM (p&lt;0.001) that was confirmed by mode size detected by NTA (p&lt;0.02). Moreover, an increased number of double and multilayer vesicles in plasma EVs from GD patients was demonstrated by cryo-EM. We also detected an increase in the expression of exosomal markers on the surface of vesicles from the blood plasma of patients with GD compared to controls. Conclusion: Here, we firstly report that the exosomes obtained from the blood plasma of GD patients have a larger size and altered morphology. Thus, we have shown that lysosomal dysfunction in GD patients leads to a striking alteration of blood plasma extracellular vesicle pool.

Characterization of soluble PD-L1 in pleural effusions of mesothelioma patients: potential implications in the immune response and prognosis.

Programmed death-ligand 1 (PD-L1) protein plays a central role in the antitumor immune response, and appears to be a predictor of prognosis and efficacy for PD-L1 and programmed death 1 (PD-1) blockade therapy. The immunoregulatory role and prognostic impact of PD-L1 soluble form (sPD-L1) have been investigated in biological fluids of patients with different tumors. In malignant pleural mesothelioma (MPM), circulating sPD-L1 has been recently reported in patients' sera, but no data are available in pleural effusions (PE). In our study, we evaluated the baseline expression levels of sPD-L1 in PE from 84 MPM patients and correlated them with PD-L1-status in matched tumors and patients' overall survival (OS). sPD-L1 in PE was determined by ELISA and tumor PD-L1 by immunohistochemistry. Association of sPD-L1 with OS was estimated using the Cox regression model. We observed that sPD-L1 was variably expressed in all the PE and tended to be higher (by 30%) in patients with PD-L1-positive tumors (cut-off ≥ 1% stained cells) as compared to patients with PD-L1-negative tumors (geometric mean ratio = 1.28, P value = 0.288). sPD-L1 levels were significantly higher than those of sPD-1 (P value = 0.001) regardless of the MPM histotypes and they were positively correlated (r = 0.50, P value < 0.001). Moreover, high PE sPD-L1 concentrations were associated with a trend towards increased OS (hazard ratio 0.79, 95% CL 0.62-1.01, P value = 0.062). Our study documents the presence of sPD-L1 in PE of MPM patients, and suggests its possible biological and prognostic role in MPM.

Serum amyloid A: A potential biomarker of lung disorders.

Serum amyloid A is an acute-phase protein with multiple immunological functions. Serum amyloid A is involved in lipid metabolism, inflammatory reactions, granuloma formation, and cancerogenesis. Additionally, serum amyloid A is involved in the pathogenesis of different autoimmune lung diseases. The levels of serum amyloid A has been evaluated in biological fluids of patients with different lung diseases, including autoimmune disorders, chronic obstructive pulmonary diseases, obstructive sleep apnea syndrome, sarcoidosis, asthma, lung cancer, and other lung disorders, such as idiopathic pulmonary fibrosis, tuberculosis, radiation pneumonitis, and cystic fibrosis. This review focuses on the cellular and molecular interactions of serum amyloid A in different lung diseases and suggests this acute-phase protein as a prognostic marker.

Neutron activation analysis for determination of metal ions in biological fluids of patients after CoCrMo arthroplasty

Determination of metals and trace elements in patients with total knee or hip arthroplasty with CoCrMo alloy was performed. Blood, urine and cerebrospinal fluid (CSF) samples were analyzed and compared with samples from healthy people. Levels of Co, Cr as well as Na, Ca, Fe, Zn, Se, Rb, Sb and Br were determined by means of neutron activation analysis. The values of Cr and Co of the blood and urine measurements were elevated in patients with replacement, and according to the statistical analysis, significant differences of the elements Zn, Br, Co and Sb were found in the CSF (p < 0.05).

Noncoding RNAs as circulating biomarkers in osteosarcoma patients.

Noncoding RNAs (ncRNAs) identify a large family of RNAs that do not encode proteins and represent an important group of tumor biomarkers, directly involved in the process of tumor pathogenesis and progression. Many of them have also been identified in biological fluids of patients with cancer, especially blood, suggesting their role as an emerging class of circulating biomarkers. Many ncRNAs, both miRNAs and lncRNAs, are deregulated in sarcoma tissues, with the most consistent data in osteosarcomas. In patients with osteosarcoma, the role of ncRNAs as circulating biomarkers is taking enormous value, above all for their ability to vary expression levels during disease progression and in response to therapy. In this mini-review, we summarize the main studies supporting the role of circulating ncRNAs in monitoring disease status in patients with osteosarcoma.

The Profile of Urinary Headspace Volatile Organic Compounds After 12-Week Intake of Oligofructose-Enriched Inulin by Children and Adolescents with Celiac Disease on a Gluten-Free Diet: Results of a Pilot, Randomized, Placebo-Controlled Clinical Trial.

The concentration of volatile organic compounds (VOCs) can inform about the metabolic condition of the body. In the small intestine of untreated persons with celiac disease (CD), chronic inflammation can occur, leading to nutritional deficiencies, and consequently to functional impairments of the whole body. Metabolomic studies showed differences in the profile of VOCs in biological fluids of patients with CD in comparison to healthy persons; however, there is scarce quantitative and nutritional intervention information. The aim of this study was to evaluate the effect of the supplementation of a gluten-free diet (GFD) with prebiotic oligofructose-enriched inulin (Synergy 1) on the concentration of VOCs in the urine of children and adolescents with CD. Twenty-three participants were randomized to the group receiving Synergy 1 (10 g per day) or placebo for 12 weeks. Urinary VOCs were analyzed using solid-phase microextraction and gas chromatography–mass spectrometry. Sixteen compounds were identified and quantified in urine samples. The supplementation of GFD with Synergy 1 resulted in an average concentration drop (36%) of benzaldehyde in urine samples. In summary, Synergy 1, applied as a supplement of GFD for 12 weeks had a moderate impact on the VOC concentrations in the urine of children with CD.

Structural Features of Amyloid Fibrils Formed from the Full-Length and Truncated Forms of Beta-2-Microglobulin Probed by Fluorescent Dye Thioflavin T.

The persistence of high concentrations of beta-2-microglobulin (β2M) in the blood of patients with acute renal failure leads to the development of the dialysis-related amyloidosis. This disease manifests in the deposition of amyloid fibrils formed from the various forms of β2M in the tissues and biological fluids of patients. In this paper, the amyloid fibrils formed from the full-length β2M (β2m) and its variants that lack the 6 and 10 N-terminal amino acids of the protein polypeptide chain (ΔN6β2m and ΔN10β2m, respectively) were probed by using the fluorescent dye thioflavin T (ThT). For this aim, the tested solutions were prepared via the equilibrium microdialysis approach. Spectroscopic analysis of the obtained samples allowed us to detect one binding mode (type) of ThT interaction with all the studied variants of β2M amyloid fibrils with affinity ~104 M−1. This interaction can be explained by the dye molecules incorporation into the grooves that were formed by the amino acids side chains of amyloid protofibrils along the long axis of the fibrils. The decrease in the affinity and stoichiometry of the dye interaction with β2M fibrils, as well as in the fluorescence quantum yield and lifetime of the bound dye upon the shortening of the protein amino acid sequence were shown. The observed differences in the ThT-β2M fibrils binding parameters and characteristics of the bound dye allowed to prove not only the difference of the ΔN10β2m fibrils from other β2M fibrils (that can be detected visually, for example, by transmission electron microscopy (TEM), but also the differences between β2m and ΔN6β2m fibrils (that can not be unequivocally confirmed by other approaches). These results prove an essential role of N-terminal amino acids of the protein in the formation of the β2M amyloid fibrils. Information about amyloidogenic protein sequences can be claimed in the development of ways to inhibit β2M fibrillogenesis for the treatment of dialysis-related amyloidosis.

DNA versus RNA oxidation in Parkinson's disease: Which is more important?

8-hydroxy-2 deoxyguanosine (8-OHdG) and the 8-hydroxyguanosine (8-OHG) are the most widely used biomarkers of nucleoside oxidation affecting DNA and RNA and are considered reliable markers of oxidative stress. Increased levels of these markers are found in the various biological fluids of patients with neurodegenerative disorders. The primary aim of our study was to assess the differences of investigated markers between patient groups and subsequently study the influence of clinical factors that might modify the levels of investigated markers during the disease progression. In this study, we analysed the 8-OHdG and 8-OHG levels in the cerebrospinal fluid (CSF) and serum from 44 patients with Parkinson's disease (PD) and 32 controls using an ELISA. There were significantly higher CSF levels of both investigated markers in Parkinson's disease patients as compared to controls (p=0.02 and p=0.04). Significantly higher CSF values of 8-OHdG were found in PD patients without dementia (p=0.05), whereas patients with dementia recorded lower 8-OHG CSF levels compared to controls (p=0.04). The disease duration and age influenced the levels of both markers within investigated groups. Oxidative DNA damage plays an important role in the early stages of PD, whereas during the progression of the disease the process is more complex, and other mechanisms are in the foreground. The measurement of 8-OHdG might be used as an "early-stage marker", whereas the decrease of 8-OHG in CSF might reflect the degree of neurodegeneration during the disease progression, suggesting its utility as a prognostic marker of advanced PD stages.

Highly selective and sensitive detection of adenosine utilizing signal amplification based on silver ions-assisted cation exchange reaction with CdTe quantum dots

Adenosine (AD) is a biomarker for monitoring the progress of lung cancer and its analysis in biological fluids of patients is significant. Here, we report a novel signal amplification platform for the sensitive and selective monitoring of AD in the biological fluids on the basis of silver ions-assisted cation exchange reaction with CdTe quantum dots (QDs). The AD aptamer was engineered to consist of two pieces of ssDNA which were respectively attached to Fe3O4 magnetic nanoparticles and CdTe QDs. In the presence of AD, it would reassemble the two pieces of ssDNA into the intact aptamer tertiary structure, and then form the stable nanoparticle sandwich structure. After magnetic separation, the final fluorescence signal with amplification process based on silver ions-assisted twice cation exchange reactions would make AD detection more sensitive, compared with directly measuring the fluorescence intensity of sandwich structure. The platform exhibits excellent sensitivity and high specificity, with a detection limit of 0.217nM for AD. Moreover, the present platform was successfully applied to the determination of AD in urine samples of lung cancer patients and the range of the recoveries was from 90％ to 106％. The results revealed that the developed platform may provide a promising analytical platform for various target molecules in the biomedical field and clinical diagnosis.

Fluorescent lipid based sensor for the detection of thymidine phosphorylase as tumor biomarker

5-Fluorouracil (5-FU) is a chemotherapic drug widely employed to treat a wide range of solid tumors. Unfortunately, it has a narrow therapeutic window and the level of its target enzymes in biological fluids of patients can vary considerably. On these premises, a new fluorescent lipid based sensor for the detection of thymidine phosphorylase, one of the target enzymes of 5-FU, was developed, to optimize patient treatment. Both cationic and anionic fluorescent liposomes containing both an amphiphile tail-tagged with a pyrene residue and a 5‐FU derivative were investigated. The effect of the presence of a bulky quencher (the bromine atom) covalently linked to the end of the alkyl chain of the anionic component on the emission signal was also evaluated. The interaction of liposomes with the target enzyme induces the occurrence of a fluorescent signal, at an extent that depends on the formulation, due to the variation of the excimer/monomer ratio. In particular, a promising specific result was obtained upon the interaction of the target enzyme with liposomes formulated with DOPC, the cationic fluorescent surfactant, the 5-FU derivative and 11-bromoundecaonic acid at 5/1/1/3 molar ratio. Langmuir compression isotherms allowed clarifying the influence of lipid organization on the response of the sensor.

Clinical presentations of Herpes Zoster Ophthalmicus (diagnosis and therapy)

To study and summarize clinical features of HZO (including the rate of complications and their nature) and to determine the relationship between clinical and laboratory data from these patients. The study included 133 patients with ophthalmic and neurological complications of HZO (group 1 (n=28) - retrospective analysis of outpatient records for the period 1995-2005; group 2 (n=95) - a prospective study for the period 2005-2015), who received a course of conservative treatment in either the Botkin City Hospital, branch № 1, or in the ophthalmic department of the Moscow herpes centre (Gerpeticheskiy Tsentr Ltd.). Laboratory tests were performed only in patients from group 2 and included: examination of biological fluids for six types of herpes viruses by polymerase chain reaction, examination of tears and urine for DNA of Chlamydia, Mycoplasma, and Ureaplasma, and serological blood testing for markers of herpes virus infection. Patients from group 1 were prescribed topical antiviral, antibacterial, and anti-inflammatory therapy, in rare cases - acyclovir per os. In group 2, the treatment included systemic antiviral medications and immune correction therapy. Anti-inflammatory therapy consisted of local and systemic non-steroidal agents (NSAIDs). The most common ophthalmic complications of HZO in both groups were stromal keratitis and keratoiridocyclitis, neurological - III and VI cranial nerves palsies. The duration of the disease in the first group ranged from 2 months to 3 years; in the second group, patients were divided into two subgroups: subgroup A with the disease duration of no more than one month (n=81) and subgroup B with the disease duration from 1.5 to 9 months (n=14). Varicella-zoster virus (VZV) DNA was present in tears and/or other biological fluids of patients from group 2 in more than 70% of cases (n=67). Particularly, in 27.4% of cases the virus was isolated in two fluids and in 7.4% of cases - in three fluids. The duration of virus production in tears and other biological fluids (saliva, blood, and urine) ranged from 10 days to 4 months. Topical non-steroidal anti-inflammatory drugs and systemic etiological treatment in case of intraocular inflammation in HZO patients may reduce the risk of severe consequences of VZV reactivation and help avoid recurrences later in life.

Evaluation of prolidase activity and oxidative stress in patients with oral lichen planus and oral lichenoid contact reactions.

The aim of this study was to evaluate prolidase activity and oxidative stress in patients with oral lichen planus (OLP) and oral lichenoid contact reactions (OLCR) using serum and salivary samples and to compare these biomarkers with each other as well as with a group of healthy subjects in order to be able to opine their role in the estimation of OLP and OLCR. Eighteen recently diagnosed patients with OLP, 32 patients with OLCR and 18 healthy controls with matched periodontal status were recruited to the study. Prolidase activity, lipid peroxidation product malondialdehyde (MDA), sialic acid (SA), and advanced oxidation protein products (AOPPs) levels in both serum and saliva were determined. Additionally, salivary flow rate and its buffering capacity were estimated. Statistical analyses were performed using the chi-square test, t-test, Mann-Whitney U-test, and Spearman's rho correlation coefficient. No statistically significant differences were observed between the study groups and the control group regarding to the basic characteristics and the periodontal status (P > 0.05). There were no statistically significant differences between OLP and OLCR groups regarding to the distribution of lesions' type, severity, and location (P > 0.05). No significant differences were found between the two study groups with regard to Prolidase activity, MDA, SA, and AOPPs (P ˃ 0.05), whereas statistically significant differences were found between the two study groups and the control group with regard to all evaluated parameters except of serum prolidase (P ˂ 0.01). Moderate correlation was found between salivary MDA and the OLP/OLCR lesion severity, whereas a weak correlation was observed between serum SA and the OLP/OLCR lesion severity (P ˂ 0.05). The findings of this study suggest an increased prolidase activity and oxidative stress and imbalance in the antioxidant defense system in biological fluids of patients with OLP and OLCR when compared with the healthy subjects. Both OLP and OLCR patients revealed almost similar prolidase activity and oxidative stress levels although these two conditions have different etiopathogenesis.