Identification of cortactin molecular forms in human urine and their possible diagnostic value

The protein composition of human urine reflects changes in the biochemical and physiological status of an individual and has an essential diagnostic value. Using precipitation/extraction methods we isolated a protein with Mr ~100 kDa in a human urine. MALDI TOF/TOF mass spectrometry identified this protein as human Src protein kinase substrate cortactin (UniProtKB/Swiss-Prot: Q14247). Screening of urine samples using Western blotting with specific anti-human cortactin antibodies revealed different proteins immunologically related to cortactin in healthy humans and patients with liver cirrhosis and lung cancer diseases. These data suggest that the level of cortacins isoform in urine might serve as a potential marker for testing acute and systemic diseases

Redox Homeostasis Index as a Criterion for Differentiated Inclusion of Antioxidants in Complex Treatment of the Patients with Liver Cirrhosis of Different Severity Degrees and Evaluation of Its Effectiveness

Introduction. The course of liver cirrhosis (LC) is determined by the appearance and increase of the severity of syntropic comorbid lesions caused by the redox homeostasis disorders with the prooxidants system activity predominance and realizing its pathogenetic mechanism through the endothelial dysfunction. Due to the important role of oxidative stress among the etiologic and pathogenetic mechanisms of the LC onset and course, the pharmacological impact on it in the complex treatment deserves priority, being one of the main “targets”. The aim of the study. To characterize the redox homeostasis index (RHI) as a criterion for the differentiated inclusion of antioxidants in the complex treatment of the patients with liver cirrhosis of different severity degree and evaluation of its effectiveness. Materials and methods. The study was conducted in three steps. After obtaining the written consent to conduct the survey, 75 patients (23 women (30.7 %), 52 men (69.3 %), mean age – 47.2 ± 10.4 years) were enrolled in a randomized trial with preliminary stratification by the presence of LC. All of them underwent the complex clinical-laboratory-instrumental examination and inpatient treatment at Lviv Regional Hepatology Center. The state of redox homeostasis was evaluated by the malondialdehyde (MDA) and catalase (CAT) levels determining, and, on the basis of the received results, we proposed to calculate RHI. Reduced RHI (18.1) – an indicator of the antioxidant system excessive activity. The first step of our study was to determine the RHI to detect the disorders of redox homeostasis in patients with different severity degrees before the treatment, i.e. obtaining the information about the presence of patients with reduced, normal and increased values, which allows us to recommend the modification of standard complex treatment using antioxidants (medicine containing retinol palmitate (vitamin A) 100 000 MO and α-tocopherol acetate (vitamin E) 0.1 g – 1 capsule per os once a day after meals daily, ascorbic acid 0.05 g – 2 tablets per os once a day after meals daily, selenium 0,0002 g – 1 tablet per os once a day after meals daily for two months) only for patients with reduced RHI. The second step of the study was devoted to the determination of RHI after a course of complex differentiated treatment of the same patients with LC of varying severity. The third step of the study was to compare the RHI obtained after the course of complex differentiated treatment of patients with LC varying severity degrees with the values of the RHI prior to treatment. The actual material was processed on a personal computer in Exсel 2010, Statistica 6.0, RStudio v. 1.1.442 and R Commander v. 2.4-4. The results obtained were presented as Me [25.0 %; 75.0 %]. The difference was considered statistically significant if p < 0.05. Results. The overwhelming majority of patients with LC (71 persons, representing 94.7 % of all those involved in the study) have disorders of redox homeostasis, among which patients with the signs of the prooxidant system prevalence and the antioxidant system activity attenuation are predominant (66 persons, accounting for 88.0 % of all those involved in the study), and the incidence of them significantly increases, as well as the value of RHI decreases with LC decompensation. Since redox homeostasis disorders with signs of attenuation of the antioxidant system activity have been diagnosed in the vast majority of patients with LC, and they determine to a large extent the features of the disease as well as the onset and severity increase of syntropic comorbid lesions, its medication adjustment is needed. RHI allows to determine the feasibility of including the antioxidants in their comprehensive treatment, as well as to determine its effectiveness, which depends on its indicators for the treatment and severity of liver cirrhosis and is the lowest in patients at the stage of decompensation of the disease. It is revealed that after the administering of the complex, differentiated standard treatment, modified by us, with the use of antioxidants RHI increases in patients with its primary decrease, which results in an improvement of the redox system state, which is the most pronounced in patients of class B, and the least – of class C by the criteria of C. G. Child – R. N. Pugh. Conclusions. The redox homeostasis index is an important criterion for assessing the state of redox homeostasis in patients with liver cirrhosis of different severity, that allows to determine the feasibility of including the antioxidants in their complex treatment, as well as to determine its effectiveness, which is the lowest in patients with decompensated liver cirrhosis

European Society of Cardiology Guidelines on the Management of Stable Coronary Artery Disease (2013)

Introduction. In 2013 the updated guidelines of the European Society of Cardiology for the treatment of stable ischemic (coronary) heart disease were published. The new document, in comparison with the same recommendations of 2006, introduced a number of significant changes that require some clarification. Aim. To consider the Recommendations of the European Cardiology Association for the management of patients with stable coronary heart disease (2013) to assist the physicians in selecting the best treatment option for the individual patient in daily practice. Materials and methods. Recommendations for application, interaction and side effects of the main drugs are considered, evaluation of possible complications in patients with ischemic (coronary) heart disease in accordance with the Recommendations of the European Association of Cardiologists regarding the management of patients with stable coronary heart disease (2013) has been made. Results. In the accordance with the recommendations, instead of the term “stable angina pectoris”, a more precise term – “stable coronary artery disease” – was used to determine the nosology. In addition, the new recommendations contain a number of changes: – in the diagnostic algorithm of the chronic heart disease more attention is paid to determining the pretest probability of the disease; – the use of up-to-date information on pretest probability will help to reduce the incidence of undesirable chronic heart disease, especially in women; – in the diagnosis of coronary artery disease, the role of visualization techniques, such as resonance imaging (MRI), computed tomography (CT), angiography, is increasing; – considerable attention is focused on the study of coronary vasospasm and microvascular dysfunction, as the most common causes of angina pectoris; – more attention is paid to combating risk factors, and therefore, prevention of serious coronary complications; – updated recommendations for the use of anti-anginal drugs are presented; – new methods of the evaluation of ischemia and damage to coronary arteries, as well as technical devices for the myocardial revascularization, are recommended; – an emphasis on the strategy of treatment: revascularization in comparison with medical treatment, as well as the choice of the method of revascularization, as necessary was made. Conclusions. The updated recommendations have the renewed information about the pathogenesis, diagnosis and treatment of stable coronary artery disease will assist the physicians in selecting the best treatment option for the individual patient in daily practice

Visual Signs of Chronic Diffuse Liver Diseases: the Stigma of Skin, its Appendages and Mucous Membranes

Introduction. The condition of the skin has always been a reflection of the state of the internal organs, in particular, organs of the gastrointestinal system, liver. Most skin symptoms are not specific to the hepatobiliary system, and therefore can be observed in other diseases. However, often the presence of skin stigma and patient complaints can be very helpful in verifying the diagnosis. According to the results of research in the world, palmar erythema occurs in 23.0%, telangiectasias (vascular “spiders”) – in 33.0%, and the triad of symptoms – palmar erythema, telangiectasias and white nails – in 21.0% of the patients with the liver cirrhosis. Despite all the importance, the topic is not sufficiently covered in the medical literature. Aim. Comprehensively describe stigmata of the skin, its appendages and mucous membranes in patients with chronic digguse liver diseases (CDLD). Materials and methods. The results of complex clinical-laboratory and instrumental examination of 2 007 patients with CDLD that were treated during the period of 2005-2013 in the Lviv Regional Hepatologic Center, which was created on the basis of the Department of Internal Medicine N 1 of the Danylo Halytsky Lviv National Medical University and the Gastroenterological Department of the Lviv Regional Clinical Hospital are presented. Among these patients there were 1 508 men (75.1%) and 499 women (24.9%) at the age of 47.9 ± 0.2 years. Before the start of treatment in a hospital, they were examined (according to the order of the Ministry of Health of Ukraine N 271 dated 13.06.05 “On Approval of Protocols of the Providing of Medical Care on the Specialty “Gastroenterology”), on the basis of the results of which the clinical diagnoses were set. Results. Skin changes are often the first predictors of CDLD. For example, a combination of pigmentation, jaundice and xanotomy confirms the diagnosis of primary liver biliary cirrhosis. In the vast majority of patients with chronic liver diseases there are pathogenetically syntropical co- and polistigmata of the skin, which are of great diagnostic value. They allow an experienced doctor to suspect or to set the diagnosis before the special comprehensive examination and to determine the tactics of treatment. The most widespread disorders found in the cirrhotic patients were dyschromia, in particular, jaundice, spider nevi, palmar erythema, «cardinal» tongue, «paper-money» skin, «caput medusae», «lacquer» nails. Conclusions. Chronic diffuse liver diseases are mainly accompanied by the pathogenic syntropic skin, its appendages and mucous membranes lesions, which are of important diagnostic value and help the experienced physician suspect or set the diagnose before a special comprehensive examination, choose the treatment tactics before the getting the results of this examination

Comorbidity: a Modern View on the Problem; Classification (first notice)

Introduction. Researches of the famous scientists proved, that comorbid diseases often worsen the course of the underlying disease and/or lead to its chronicity, cause disability and premature death of the working population, often make inefficient diagnosis and treatment, increase the costs of the health system to provide medical care. According to the research of WHO experts, people under 40 usually have 2-4 diseases, under 60 – there is a “bouquet” of 5-7 diseases, after 70 – 8-10 or more diseases. So-called “polymorbidity” increases from 10.0 % in patients aged 80 years old. Materials and research methods. The work analyzes the literature sources, which deal with the study of comorbid diseases and sufficiently thoroughly reflect the current views of the scientists on the issue. Results of the investigation and their discussion. There are many synonyms of the term “comorbidity”, among them, often are used the following: “polymorbity”, “multimorbidity”, “multifactorial diseases”, “polypathy”, “multicausal diagnosis”, “dual diagnosis”, “pluripathology” etc., which complicates the understanding of the problem and minimizes the ability to use scientific achievements in clinical practice. In Ukrainian literature the authors often use the term “bicausal diagnosis” to describe comorbidity when there are two underlying diseases, to describe polymorbidity – “multicausal diagnosis” (three or more pathological conditions in one individual). In English scientific literature the authors often use the terms “comorbid diseases”, “comorbid conditions”. In general, the sources of information, used by the researchers to study the problem of polypathies, were and are medical histories, hospital records of the patients and other medical records of family doctors, of insurance companies, archives of the pensions for the elderly. Comorbidity itself can be interpreted as a random combination of the diseases, different by its’ etiology and pathogenesis such as nosological syntropy, ie natural occurrence of the regular determined diseases combinations in one patient. The main causes of comorbidity are: affected organs anatomical proximity, common pathogenesis, causal connections, combination of the diseases or iatrogenic causes. Among the factors, that influence the occurrence of comorbidity in humans, is a chronic infection, inflammation, involutive and systemic metabolic changes, iatrogenic factors, social status, environment and genetic predisposition, and the main way of the development of comorbid disorders is causal. The appearing of a particular disease leads to functional first, and later – to organic lesions and further – to an increase of nosologies quantity. Despite the large number of studies, proposed definitions and synonyms, standardized terminology and conventional classification of comorbidity, that would greatly help in the study of its problems, until now is missing. Conclusions. Despite many unsolved patterns of comorbidity and lack of the unified terminology, the search for the new variants of combinations of these diseases continues. After all, they can simply be not yet classified syndromal diseases, or vice versa – to be completely incompatible for the appearance and coexistence in one body. This is the key to solve more and more problems for the diagnosis and treatment of the diseases in the field of the practical medicine

State of the Autonomic Nervous System in Patients with the Different Severity Stages of Cirrhosis of the Liver

Introduction. A problem of the occurrence and course of liver cirrhosis (LC) and its severity increase is one of the most difficult in modern gastroenterology. An important place in the pathogenesis of LC, the course of the disease and its prognosis takes the state of the autonomic nervous system (ANS). It is known that the disorders of the ANS state is one of the pathogenetic mechanisms of the appearance and increase of the severity of LC, due to the exclusion of the central regulatory mechanisms of life processes with the transition to the lowest level – humoral-metabolic, which is an unfavorable prognostic sign. However the features of ANS state f, in spite of its important role in the human organism, are not sufficiently studied. Aim. To find out the ANS state in cirrhotic patients with the different degree of the severity. Materials and methods. In order to assess the state of the ANS we have conducted a comprehensive clinical and laboratory examination of 93 patients with LCand studied its basic functional and dynamic indices: tone, autonomic reactivity, autonomic support of the organism functioning, using the questionnaire of A. M. Wein, clinical criteria of A. D. Solovyov, functional tests: thermal, cold, Tom-Roux, orthoclinostatic and registration of heart rate variability. Results. According to the results of A. M. Wein’s questionnaire, the most often symptoms of the autonomic disorders were met in the patients of C class (by C. G. Child-R. N. Pugh). According to the results of the medical examination, in obedience to the criteria of A. D. Solovyov, the tone of the sympathetic nervous system (SNS) prevailed in all groups of the patients: for the persons of class A – on 30.0 %, B – on 44.0 %, C – on 51.6 % and for the control group (CG) – on 3.0 %, comparing with the tone of the parasympathetic nervous system (PSNS). Consequently, the conducted studies have allowed us to diagnose the disorders of ANS in all the patients with LC, severity of which significantly increases with the disease decompensation. In all the people SNS tone has prevailed with a tendency to hypertone with increasing of the pathological process severity, indicating the activation of compensatory capacity of the organism to maintenance homeostasis in case of portal hypertension. Results, received after the conducting of cold test, testify, that comparatively to the CG, in most persons of class A (91.3 %) hyperreactivity of the ANS is diagnosed more frequently (p < 0.001), that is in 74.5 and 91.3 % of the persons of classes B and C accordingly (p < 0.001). The results of thermal, orthoclinostatic, Tom-Roux’s probes confirm the indices, got during the conducting of the cold test. The results of the thermal, orthoclinostatic and Tom- Roux’s tests were confirmed by the indices, received during the cold test. Also, during the record of the heart rate variability without stress the indices of the general spectral power (TP), in patients of class A (923.4 ± 191.3 mc2/Hz), B (935.9 ± 139.4 mc2/Hz) and C (203.4 ± 301.9 mc2/Hz) were significantly (p < 0.001) lower than in the CG (3243.8 ± 46.2 mc2/Hz). In the patients of class C the indices of TP were significantly lower comparing with class A (p < 0.001) and B (p < 0.001) with the tendency to the decrease with the increase of severity class. It testifies the strengthening of the SNS influence, however not reflect the structure of the organism regulation levels. The results, got during the conducting of the orthostatic test, confirmed the indices, got before the stress. Conclusions. The results allowed us to diagnose the presence of autonomic disorders in 100.0 % of the patients, with the superior effects of the sympathetic part of the autonomic nervous system over the parasympathetic with the directly proportional increase of the neuro-humoral effects according to the severity of the pathological process in the liver. The study of the correlation between the indices of heart rate variability and classes of liver cirrhosis by C. G. Child-R. N. Pugh lets us recommend them to predict the disease course

Severity Degrees of Syntropic Stable Arterial Hypotension in the Cirrhotic Patients and Their Dependence on the Daily Blood Pressure Monitoring and Liver Damage Class by C.G. Child – R.N. Pugh

Introduction. Daily monitoring of the arterial blood pressure in the cirrhotic patients is used in the practical medicine, as well as in numerous scientific studies for the study of the pathogenesis, clinical and instrumental manifestations of the circulatory system injury. Daily monitoring of the arterial blood pressure should also be used for the syntropic arterial hypotension severity degrees gradation inthe cirrhotic patients. The aim of the study. To determine the syntropic arterial hypotension severity degrees in the cirrhotic patients, their dependence on the daily blood pressure monitoring and liver injury class according to C.G. Child – R. N. Pugh. Materials and methods. The study involved 603 patients with liver cirrhosis (445 men and 158 women, mean age – 48.4 ± 10.3 years), among which – 173 patients with syntropic arterial hypotension, 89 patients of which – with syntropic arterial hypotension without any other disorders of the circulatory system (experimental group). The control group included 26 practically healthy volunteers of the respective gender and age. The daily blood pressure monitoring using the ABPM-04 (Meditech, Hungary) device according to the standard procedure was conducted for all the patients, in addition to the usual examination procedure. Results. It was found that the average, maximum and minimum daily systolic, diastolic, mean and pulse blood pressure, as well as their derivative parameters in the patients of the experimental group, compared with their values in the practically healthy volunteers, are significantly lower, which confirms the presence of stable arterial hypotension in such category of patients. The disorders of the circadian rhythm and blood pressure variability were also found in the patients of the experimental group. The average daily blood pressure is advisable to use to determine the severity of hypotension. In case if the average daily blood pressure, determined by the method of daily monitoring, ranges from 80 mmHg to 76 mmHg, we diagnose mild (I) degree of hypotension, from 75 mmHg to 70 mmHg – the average (II), and in cases when the average daily blood pressure is less than 70 mm Hg -severe (III) degree of arterial hypotension. Among the examined patients (173 patients with arterial hypotension), 54 patients with I degree (31.21 %), 66 patients with II (38.15 %) and 53 patients with III (30.64 %) degree of arterial hypotension were found. The study of correlation between the indicators of daily blood pressure monitoring, depending on the arterial hypotension severity, showed that the average daily blood pressure is the most informative parameter, also the number of some other indicators are informative. After analyzing the distribution of degrees of arterial hypotension, depending on the severity of livercirrhosis by C.G. Child and R.N. Pugh, we found that with the increase of the class, the number of patients with mild (I degree) hypotension decreases and the number of patients with secondary (II degree) and severe (III degree) arterial hypotension increases. Conclusions. We characterized the arterial pressure, recorded by the method of daily blood pressure monitoring. It was found that in the cirrhotic patients there is a syntropic stable arterial hypotension with the disturbed circadian rhythms and arterial pressure variability. The average daily blood pressure is set as the most informative parameter, and the syntropic hypotension is classified into three degrees of severity. Correlative relationships between the pa­rameters of daily blood pressure monitoring and degrees of severity of stable arterial hypotension, among which the strongest has an average daily blood pressure, are revealed. It was stated that with the increase of the liver cirrhosisseverity class by C. G. Child and R. N. Pugh, the number of patients with mild degree syntropic hypotension decreases and the number of patients with syntropic arterial hypotension of moderate andsevere degrees increases

Screening of riboflavin kinase activity in microorganisms

The riboflavin kinase activity was measured in different species of bacteria, yeasts and molds. The activity of enzyme in eucaryotic microorganisms was mainly higher comparing to procaryotes. It was shown that bacterial and yeasts riboflavin kinases of investigated strain besides riboflavin can utilize its reduced form – 1,5 dihydroriboflavin as a substrate. Quantum chemical calculation of three-dimentional model of structure of oxidized and reduced form of riboflavin showed substantial differences in their configuration. No correlation between the activity of riboflavin kinase and dihydroriboflavin kinase in obligate aerobes and microaerophiles was observed

Adaptation of Conductometric Monoenzyme Biosensor for Rapid Quantitative Analysis of L-arginine in Dietary Supplements

The present study reports on the development, adaptation, and optimization of a novel monoenzyme conductometric biosensor based on a recombinant arginine deiminase (ADI) for the determination of arginine in dietary supplements with a high accuracy of results. Aiming for the highly sensitive determination of arginine in real samples, we studied the effect of parameters of the working buffer solution (its pH, buffer capacity, ionic strength, temperature, and protein concentration) on the sensitivity of the biosensor to arginine. Thus, it was determined that the optimal buffer is a 5 mM phosphate buffer solution with pH 6.2, and the optimal temperature is 39.5 °C. The linear functioning range is 2.5–750 µM of L-arginine with a minimal limit of detection of 2 µM. The concentration of arginine in food additive samples was determined using the developed ADI-based biosensor. Based on the obtained results, the most effective method of biosensor analysis using the method of standard additions was chosen. It was also checked how the reproducibility of the biosensor changes during the analysis of pharmaceutical samples. The results of the determination of arginine in real samples using a conductometric biosensor based on ADI clearly correlated with the data obtained using the method of ion-exchange chromatography and enzymatic spectrophotometric analysis. We concluded that the developed biosensor would be effective for the accurate and selective determination of arginine in dietary supplements intended for the prevention and/or elimination of arginine deficiency

Efficient production of bacterial antibiotics aminoriboflavin and roseoflavin in eukaryotic microorganisms, yeasts

Abstract Background Actinomycetes Streptomyces davaonensis and Streptomyces cinnabarinus synthesize a promising broad-spectrum antibiotic roseoflavin, with its synthesis starting from flavin mononucleotide and proceeding through an immediate precursor, aminoriboflavin, that also has antibiotic properties. Roseoflavin accumulation by the natural producers is rather low, whereas aminoriboflavin accumulation is negligible. Yeasts have many advantages as biotechnological producers relative to bacteria, however, no recombinant producers of bacterial antibiotics in yeasts are known. Results Roseoflavin biosynthesis genes have been expressed in riboflavin- or FMN-overproducing yeast strains of Candida famata and Komagataella phaffii. Both these strains accumulated aminoriboflavin, whereas only the latter produced roseoflavin. Aminoriboflavin isolated from the culture liquid of C. famata strain inhibited the growth of Staphylococcus aureus (including MRSA) and Listeria monocytogenes. Maximal accumulation of aminoriboflavin in shake-flasks reached 1.5 mg L− 1 (C. famata), and that of roseoflavin was 5 mg L− 1 (K. phaffii). Accumulation of aminoriboflavin and roseoflavin by K. phaffii recombinant strain in a bioreactor reached 22 and 130 mg L− 1, respectively. For comparison, recombinant strains of the native bacterial producer S. davaonensis accumulated near one-order less of roseoflavin while no recombinant producers of aminoriboflavin was reported at all. Conclusions Yeast recombinant producers of bacterial antibiotics aminoriboflavin and roseoflavin were constructed and evaluated