[Targeting NMDAR/AMPAR: a promising pharmacotherapeutic approach for depressive disorders]. / Targetirovanie NMDAR/AMPAR ­ perspektivnyi podkhod farmakokorrektsii depressivnykh rasstroistv.

Depression is a leading cause of disability and reduced work capacity worldwide. The monoamine theory of the pathogenesis of depression has remained dominant for many decades, however, drugs developed on its basis have limited efficacy. Exploring alternative mechanisms underlying this pathology could illuminate new avenues for pharmacological intervention. Targeting glutamatergic pathways in the CNS, particularly through modulation of NMDA and AMPA receptors, demonstrates promising results. This review presents some existing drugs with glutamatergic activity and novel developments based on it to enhance the efficacy of pharmacotherapy for depressive disorders.

[Evolution of insulin therapy: past, present, future].

2021 marks the 100th anniversary of the discovery of insulin, an event that forever changed the lives of people with diabetes mellitus. At present patients around the world experience the miracle of insulin therapy every day. A disease that used to kill children and teenagers in 2 years in 1920 has become a disease that can be controlled with a possibility to lead a long productive life. Over the past century, the great discovery of Banting, Best and Collip has forever changed the world and saved millions of lives. This review is devoted to the history of the development of insulin and its further improvement: from the moment of discovery to the present days. Various generations of insulin are considered: from animals to modern ultrashort and basal analogues. The article ends with a brief review of current trends in the development of new delivery methods and the development of new insulin molecules. Over the past century, insulin therapy has come a long way, which has significantly improved the quality of life of our patients. But research is actively continuing, including in the field of alternative methods of insulin delivery, which are more convenient for the patient, as well as in the development of «smart¼ molecules that will have a glucose-dependent effect.

[First experience of applying domestic video fluorescent equipment for visualization and blood flow evaluation of the parathyroid glands]. / Pervyi opyt primeneniya otechestvennoi videoflyuorestsentnoi ustanovki dlya vizualizatsii i otsenki krovotoka okoloshchitovidnykh zhelez.

In recent years, predictive methods for assessing the preservation of the parathyroid glands have been actively implemented. The article describes the first experience of evaluating the blood supply of the parathyroid glands by quantitative determination of the indocyanine green (ICG) accumulation index in real time in 6 patients before and after a thyroidectomy with central neck lymph node dissection for papillary thyroid cancer. Intraoperative fluorescent angiography was performed by using domestic equipment with a fluorescent module, as well as by using a domestic medication of ICG. Intraoperative values of the ICG accumulation index were compared with the levels of ionized calcium and parathyroid hormone perioperatively. No clinical manifestations of hypocalcemia were detected in the postoperative period. The obtained results showed the informativeness of the numerical assessment of the intensity of ICG fluorescence. The evaluation of the distribution (accumulation) of ICG has prospects for practical application in thyroid surgery in the formation of tactics for preserving the parathyroid glands and predicting postoperative hypoparathyreosis.

Combining branched copolymers with additives generates stable thermoresponsive emulsions with in situ gelation upon exposure to body temperature.

Branched copolymer surfactants (BCS) containing thermoresponsive polymer components, hydrophilic components, and hydrophobic termini allow the formation of emulsions which switch from liquid at room temperature to a gel state upon heating. These materials have great potential as in situ gel-forming dosage forms for administration to external and internal body sites, where the emulsion system also allows effective solubilisation of a range of drugs with different chemistries. These systems have been reported previously, however there are many challenges to translation into pharmaceutical excipients. To transition towards this application, this manuscript describes the evaluation of a range of pharmaceutically-relevant oils in the BCS system as well as evaluation of surfactants and polymeric/oligomeric additives to enhance stability. Key endpoints for this study are macroscopic stability of the emulsions and rheological response to temperature. The effect of an optimal additive (methylcellulose) on the nanoscale processes occurring in the BCS-stabilised emulsions is probed by small-angle neutron scattering (SANS) to better comprehend the system. Overall, the study reports an optimal BCS/methylcellulose system exhibiting sol-gel transition at a physiologically-relevant temperature without macroscopic evidence of instability as an in situ gelling dosage form.

Structuring and de-structuring of nanovectors from algal lipids. Part 1: physico-chemical characterization.

Lipid nanocarriers are among the most employed systems for drug delivery purposes in several research and industrial sectors, since their favorable properties ensure broad applicability. The design and characterization of these nanosystems are of paramount importance to obtain controlled outcome, since the supramolecular structure and molecular interactions deeply impact the functionality of the resulting aggregates. The choice of the most appropriate formulation for the target of interest relies on in-depth physico-chemical characterization in order to optimize stability, loading rates and sustained release. Several supramolecular architectures suited for carrier development can be obtained from lipid building blocks, by varying lipid composition and packing parameter. In particular, cubosome and liposome aggregates are often used as drug vectors thanks to their high cargo capability and biocompatibility. Moreover, the possibility to employ lipids from natural sources i.e. biomasses to prepare nanosystems makes them especially attractive. In this work, two aggregate types were characterized and compared as drug vectors for poorly water-soluble antioxidants, particularly curcumin and two adjuvants (i.e. tocopherol and piperine). The nanovectors were obtained by extracting lipids from algal biomasses with different lipid composition, and characterized by advanced structural (DLS, SAXS, Cryo-TEM) techniques, spectroscopy (NMR) and calorimetry (ITC). Finally, the structural stability of both aggregate types was evaluated.

Transcranial direct current stimulation of the premotor cortex aimed to improve hand motor function in chronic stroke patients.

OBJECTIVE: To investigate the effects of single-session premotor and primary motor tDCS in chronic stroke patients with relation to possible inter-hemispheric interactions. METHODS: Anodal tDCS of either M1 or premotor cortex of the side contralateral to the paretic hand, cathodal tDCS of the premotor cortex of the side ipsilateral to the paretic hand and sham stimulation were performed in 12 chronic stroke patients with mild hand paresis in a balanced cross-over design. The Jebsen-Taylor Hand Function test, evaluating the time required for performance of everyday motor tasks, was employed. RESULTS: The repeated-measure ANOVA with Greenhouse-Geisser correction showed significant influence of the stimulation type (factor SESSION; F(2.6, 28.4) = 47.3, p < 0.001), the test performance time relative to stimulation (during or after tDCS; factor TIME, F(1.0, 11.0) = 234.5, p < 0.001) with higher effect after the stimulation and the interaction SESSION*TIME (F(1.7, 1.2) = 30.5, p < 0.001). All active conditions were effective for the modulation of JTT performance, though the highest effect was observed after anodal tDCS of M1, followed by effects after anodal stimulation of the premotor cortex contralateral to the paretic hand. Based on the correlation patterns, the inhibitory input to M1 from premotor cortex of another hemisphere and an excitatory input from the ipsilesional premotor cortex were suggested. CONCLUSION: The premotor cortex is a promising candidate area for transcranial non-invasive stimulation of chronic stroke patients.

A Monoiodotyrosine Challenge Test in a Parkinson's Disease Model.

Early (preclinical) diagnosis of Parkinson's disease (PD) is a major challenge in modern neuroscience. The objective of this study was to experimentally evaluate a diagnostic challenge test with monoiodotyrosine (MIT), an endogenous inhibitor of tyrosine hydroxylase. Striatal dopamine was shown to decrease by 34% 2 h after subcutaneous injection of 100 mg/kg MIT to intact mice, with the effect not being amplified by a further increase in the MIT dose. The selected MIT dose caused motor impairment in a neurotoxic mouse model of preclinical PD, but not in the controls. This was because MIT reduced striatal dopamine to the threshold of motor symptoms manifestation only in PD mice. Therefore, using the experimental mouse model of preclinical PD, we have shown that a MIT challenge test may be used to detect latent nigrostriatal dysfunction.

[Development of early diagnosis of Parkinson's disease and comprehensive economic analysis of the effect of its implementation]. / Razrabotka rannei diagnostiki bolezni Parkinsona i kompleksnyi ekonomicheskii analiz effekta ot ee vnedreniya.

The paper summarizes the literature and author's data on the development of early (preclinical) diagnosis of Parkinson's disease (PD). Implementation of this diagnosis will promote the use of preventive therapy and change investments in diagnosis and treatment of patients. The paper declares that at present the only approach to early diagnosis of PD is positron-emission tomography of the nigrostriatal dopaminergic system, but it cannot be used for preventive examination due to its high cost. The authors consider that a less specific, but more promising approach to the development of early diagnosis of PD is the search for markers in body fluids, mainly in the blood, in patients at the prodromal stage of PD. Indeed, a number of markers as changes in the level of metabolites of monoamines, sphingolipids, urates, and indicators of oxidative stress were found in patients selected for the risk group of the prodromal stage of PD, according to characteristic premotor symptoms. In addition, it is assumed that the search for blood markers at an earlier - pre-prodromal stage is possible only in animal models of PD at the early preclinical stage. This approach can also be used to verify blood markers identified in patients at the clinical stage of PD. It is also evident that the complex socio-economic factors influencing the incidence of PD is different in developed versus developing countries. The societal and medical costs of Parkinson's are huge and efforts to improve early preclinical diagnosis of PD will lead to considerable economical and societal benefits. For instance this will allow efficient selection of patients for preclinical diagnostic tests. To assess the effectiveness of this strategy considering the uncertainty of socio-economic issues, a modification of the «cost-utility¼ analysis is proposed. For the first time, a Markov model of PD including preclinical diagnostic tests and possible neuroprotective therapy was developed and studied. Analytical outcomes of this process suggest that the idea of developing a new multimodal strategy is promising from a socio-economic point of view.

Polyethylenimine based magnetic nanoparticles mediated non-viral CRISPR/Cas9 system for genome editing.

Clustered regularly interspaced short palindromic repeats-associated protein (CRISPR/Cas9) system has become a revolutionary tool for gene editing. Since viral delivery systems have significant side effects, and naked DNA delivery is not an option, the nontoxic, non-viral delivery of CRISPR/Cas9 components would significantly improve future therapeutic delivery. In this study, we aim at characterizing nanoparticles to deliver plasmid DNA encoding for the CRISPR-Cas system in eukaryotic cells in vitro. CRISPR/Cas9 complexed polyethylenimine (PEI) magnetic nanoparticles (MNPs) were generated. We used a stable HEK293 cell line expressing the traffic light reporter (TLR-3) system to evaluate efficient homology- directed repair (HDR) and non-homologous end joining (NHEJ) events following transfection with NPs. MNPs have been synthesized by co-precipitation with the average particle size around 20 nm in diameter. The dynamic light scattering and zeta potential measurements showed that NPs exhibited narrow size distribution and sufficient colloidal stability. Genome editing events were as efficient as compared to standard lipofectamine transfection. Our approach tested non-viral delivery of CRISPR/Cas9 and DNA template to perform HDR and NHEJ in the same assay. We demonstrated that PEI-MNPs is a promising delivery system for plasmids encoding CRISPR/Cas9 and template DNA and thus can improve safety and utility of gene editing.

[Development of early diagnosis of Parkinson's disease based on the search for biomarkers such as premotor symptoms and changes in blood]. / Razrabotka rannei diagnostiki bolezni Parkinsona na osnove biomarkerov v vide premotornykh simptomov i izmenenii v krovi.

OBJECTIVE: To determine changes in the chemical composition of blood plasma in subjects at risk of Parkinson's disease (PD) at the prodromal stage compared with age control. MATERIAL AND METHODS: Subjects at risk were selected for the presence of characteristic premotor symptoms, including impairments of sleep, olfaction and constipation.The risk group included 12 people, the control group - 8 people. RESULTS: Among seven catecholamines and their metabolites detected in the blood, only the concentration of L-dioxiphenylalanine (L-DOPA) changed (decreased) in subjects at risk compared with the control. A decrease in the concentration of L-DOPA is considered as a manifestation (marker) of selective degeneration of central and peripheral catecholaminergic neurons in PD. In contrast to L-DOPA, the concentration of seven of the twelve detected sphingomyelins in the blood of the subjects at risk increased. Given that a change in the metabolism of sphingomyelins is associated with processes such as apoptosis, autophagy, and synucleinopathy, an increase in their concentration in the blood of patients at risk is considered as a manifestation of systemic general degeneration of central and peripheral neurons. Finally, in the blood of subjects at risk, we found a trend towards a decrease in the concentration of urates, which are endogenous neuroprotectors. CONCLUSION: The changes in the level of L-DOPA, sphingmyelins and urates in the blood of subjects at risk may serve as diagnostic markers of PD at the prodromal stage.

[Transcranial direct current stimulation in neurology and psychiatry]. / Transkranial'naya stimulyatsiya postoyannym tokom v nevrologii i psikhiatrii.

Transcranial direct current stimulation (tDCS) is a non-invasive method of modulating brain excitability by low intensity direct current. At present, there are numerous studies of tDCS application in various mental and neurological diseases. In this review, the data of tDCS efficiency in the treatment of different disorders are presented and the recommendations on using this method in clinical practice are given.

Effect of tDCS on Fine Motor Control of Patients in Subacute and Chronic Post-Stroke Stages.

In this study we compared the effects of transcranial direct current stimulation (tDCS) in the subacute and chronic stages of post-stroke recovery. Anodal/sham tDCS was applied to the primary motor cortex of stroke patients in these stages of recovery in a cross-over design. The Jebsen-Taylor hand function test was employed. The repeated-measure ANOVA showed significant influence of the stimulation type and test performance time (during/after tDCS) with no overall influence of recovery stage. The interaction TYPE*TIME*STAGE was significant. The effect after anodal tDCS in the subacute stage was significantly higher compared to the effects in all relevant conditions including the chronic stage. Therefore, tDCS treatment in the subacute stage of recovery can be superior, at least for some patients, to treatment in the chronic stage.

Medicinal leech antimicrobial peptides lacking toxicity represent a promising alternative strategy to combat antibiotic-resistant pathogens.

The rise of antibiotic resistance has necessitated the development of alternative strategies for the treatment of infectious diseases. Antimicrobial peptides (AMPs), components of the innate immune response in various organisms, are promising next-generation drugs against bacterial infections. The ability of the medicinal leech Hirudo medicinalis to store blood for months with little change has attracted interest regarding the identification of novel AMPs in this organism. In this study, we employed computational algorithms to the medicinal leech genome assembly to identify amino acid sequences encoding potential AMPs. Then, we synthesized twelve candidate AMPs identified by the algorithms, determined their secondary structures, measured minimal inhibitory concentrations against three bacterial species (Escherichia coli, Bacillus subtilis, and Chlamydia thrachomatis), and assayed cytotoxic and haemolytic activities. Eight of twelve candidate AMPs possessed antimicrobial activity, and only two of them, 3967 (FRIMRILRVLKL) and 536-1 (RWRLVCFLCRRKKV), exhibited inhibition of growth of all tested bacterial species at a minimal inhibitory concentration of 10 µmol. Thus, we evidence the utility of the developed computational algorithms for the identification of AMPs with low toxicity and haemolytic activity in the medicinal leech genome assembly.

Proteins of the Vesicular Cycle as a Marker of Neuroplasticity of Dopaminergic Neurons in the Substantia Nigra of the Brain.

Nigrostriatal dopaminergic neurons (DNs), involved in the regulation of motor function, are characterized by a high plasticity. Indeed, at the death of up to 50% of DNs in Parkinson's disease, the survived neurons provide normal regulation. This study was aimed to determine whether the vesicle cycle proteins, syntaxin Ia (Syn Ia), synaptotagmin I (Syt I), Rab5a, and complexins I and II (Cmpx I and II) are involved in the mechanisms of neuroplasticity in the substantia nigra, which mainly contains cell bodies and processes of the DNs. In the neurotoxic models of Parkinson's disease in mice, it was shown that, at the degeneration of up to 50% of DNs, the content of Syt I, Syn Ia, and Cmpх I and II, involved in vesicle exocytosis, does not change in the substantia nigra as a whole but is compensatorily increased in individual survived DNs. Thus, the data obtained in this study suggest that the impairment of motor behavior, which occurs at the death of half of the nigrostriatal DNs, is not caused by the impairment of the production of vesicle cycle proteins in the survived DNs.

[A comparative hormonal and clinical analysis of thyrotoxicosis with- or without comorbid resistant depression].

AIM: To compare clinical and hormonal status in thyrotoxicosis with- or without comorbid resistant depression. MATERIAL AND METHODS: We examined 100 patients with thyrotoxicosis, including 61 patients with comorbid resistant depression (RD) and 39 patients without RD. Mental status, somatic symptoms of disease and blood content of main thyroid hormones were studied. The Hamilton scale was used to measure depression and anxiety. RESULTS AND CONCLUSION: Depression developed in the structure of thyrotoxicosis increased the symptoms of hyperthyroidism. However, it did not change the general tendency of development of the main disease assessed by hormonal indicators.

[ABERRANT RIGHT SUBCLAVIAN ARTERY (ARSA)--A NEW ULTRASOUND MARKER FOR CHROMOSOMAL FETAL ABNORMALITIES].

UNLABELLED: The development of the fetal aorta ends with the formation of the aortic arch which normally branches into three blood vessels: 1) a. brachiocephalica (a. innominata), which divides into the right subclavian artery (RSA) and the right carotid artery; 2) the left carotid artery; and 3) the left subclavian artery. Occasionally, RSA originates as a separate fourth branch of the aortic arch, passing behind the trachea with an oblique course to the right shoulder. This rare variant is called an aberrant right subclavian artery (ARSA) and is observed in approximately 2% of normal individuals. On the other hand, the reported incidence of ARSA varies between 25 and 37% in cases with Down syndrome and other chromosomal abnormalities. OBJECTIVE: To evaluate the success rate of ultrasound visualization of the fetal RSA between 18 and 23 weeks of gestation and to establish the importance of the prenatal diagnosis of ARSA in the risk assessment for fetal chromosomal abnormalities in the second trimester. RESULTS: Three experienced sonographers scanned 992 fetuses in MC "Markovs", Sofia between 01.09.2013-01.06.2014 with Voluson 730 Expert (GE Healthcare) ultrasound equipment. Visualization of RSA was successful in 92.7% of cases. Overall, 17 cases with ARSA were diagnosed in the study period. ARSA was an isolated sonographic finding in 13 of them. The remaining 4 cases had additional pathology. In the first case ARSA was associated with a short femurand humerus, short nasal bone and borderline nuchal thickness without any other soft markers or structural abnormalities. Trisomy 21 was diagnosed after amniocentesis and the pregnancy was terminated at patient's request. In the second case ARSA was associated with severe polymalformation syndrome. Trisomy 18 was diagnosed by DNA analysis post abortem. In the third case ARSA was associated with an unilateral cleft lip and cleft palate. Abnormalities of the fetal karyotype and Di George syndrome were excluded by amniocentesis. The fourth case was associated with a single umbilical artery without any structural and chromosomal abnormalities. The pregnancy had a favourable perinatal outcome at term. CONCLUSION: Visualization of RSA and prenatal diagnosis of ARSA in the second trimester is relatively easy in experienced hands. The examination slightly prolongs the fetal morphology scan. Since there is an obvious association between ARSA and chromosomal fetal abnormalities, implementation of its sonographic evaluation in the protocol of fetal echocardiography in the second trimester is strongly recommended.

[THE IMPACT OF 3D ULTRASOUND ON MATERNAL-FETAL BONDING DURING THE FETAL ANOMALY SCAN IN THE FIRST AND SECOND TRIMESTER].

OBJECTIVES: To assess the role of prenatal conventional (2D) and volume (3D/4D) ultrasound on maternal-fetal bonding and to investigate its emotional impact on women's perceptions. METHODS: Overall 133 pregnant women with a mean maternal age 29.81 ± 5.56 years presented for a routine first and second trimester scan in MC "Markovs" and University hospital of Obstetrics and Gynecology "Maichin dom", Sofia, Bulgaria. All participants had uncomplicated singleton pregnancies with no fetal abnormalities. The women were asked to complete a questionnaire in two parts. The first part assessed the patient's understanding and knowledge about the different imaging modalities (2D and 3D), the level of maternal-fetal attachment and the overall expectations about the scan. The second part of the questionnaire was completed after the examination and assessed the expectations of the mode of visualization, the emotional perceptions of the fetus and the maternal-fetal attachment. RESULTS: Maternal-fetal bonding increased after both the 2D and 3D/4D ultrasound examination. However, almost half of the pregnant women did not understand the difference between the two imaging modalities. There was no accumulative effect of prenatal ultrasound on maternal-fetal bonding in late gestation. Most of the patients declared that the scan had improved their overall perception of the fetus. Maternal age, educational status and gestational age had no significant impact on understanding and improving womens' emotional perceptions. CONCLUSIONS: Prenatal 2D and 3D/4D ultrasound has a positive impact on antenatal emotional maternal-fetal bonding, particularly when performed in the first trimester.

[THE FETAL MIDDLE CEREBRAL ARTERY PEAK SYSTOLIC VELOCITY AS A PEDICTOR OF FETAL ANEMIA IN RH-ALLOIMMUNIZED PREGNANCY].

UNLABELLED: Rh-isoimmunization is a pathological condition in which the fetal red blood cells of a Rh (+) fetus are destroyed by the isoantibodies of a Rh (-) woman sensitized in a previous event. Despite of the wide spread implementation of anti D-gammaglobolin prophylaxis this is still the most common cause for fetal anemia. Recently, sonographic measurement of the fetal middle cerebral artery peak systolic velocity (MCA-PSV) has been shown to be an accurate non-invasive test to predict low fetal hemoglobin levels. We present a case report of Rh-alloimmunized pregnancy with moderate fetal anemia, followed-up by weekly MCA-PSV measurements. CASE REPORT: A 37-year-old Rh (-) negative gravida 3, para 1, without anti-D gammaglobolin prophylaxis in her previous pregnancies, presented at 27+0 weeks of gestation (w.g.) for a routine third trimester scan. Subsequent ultrasound measurements of MCA-PSV confirmed a progressive increase of the peak systolic velocities from 40 to 80 cm/sec, as well as a gradual rise in the anti-D titers. The evidence of developing fetal anemia necessitated elective Caesarean section performed at 35 wg. The neonate was admitted in the intensive care unit and required resuscitation, one exchange blood transfusion and several courses of phototherapy. The patient was discharged two weeks post partum. CONCLUSIONS: There is a strong correlation between the high peak systolic velocities in the middle cerebral artery (MCA-PSV) and the low levels of fetal hemoglobin. The high sensitivity and positive predictive value concerning the development of fetal anemia, as well as its good repeatability, makes this non-invasive test a valuable asset in the management of all pregnancies complicated by severe Rh-alloimmunization.

[Three-dimensional (3D) ultrasound in evaluation of fetal brain anatomy].

OBJECTIVES: The objective of this study is to evaluate the potential of three-dimensional (3D) ultrasound identifying the fetal brain structures during second trimester and the time of offline analysis of brain volumes to perform a basic and a detailed fetal neurosonogram. METHODS: 140 pregnant women were included in this study presented for a second trimester routine ultrasound scan in Medical Centre "Markovs", Sofia and the University Hospital in Obstetrics and Gynecology, Maichin dom, Sofia, Bulgaria. All pregnancies were singleton with no diagnosed fetal anomalies. 3D fetal brain volumes were obtained by transabdominal aquisition in the most suitable plane and analyzed later offline by two trained ultrasonologists with special software. RESULTS: A standart systematic volume orientation was used to identify a list of brain structures and to complete basic evaluation, intracranial biometry and elected structures for detailed fetal neurosonogram. Diagnostic visualization was possible with excellent visualization rate in 91-100% in all brain structures for basic evaluation and in 76-100% for detailed targeted neurosonogram. An average number of 7 volumes was used to complete the examination by the two specialists. In this study a few limitation of the method are presented. CONCLUSION: 3D volume ultrasound is useful method for identification of fetal brain structures in second trimester. There are advantages in storage the digital information and for education.

[Three- and four-dimensional (3D/4D) ultrasound in obstetric practice: review].

Three-dimensional (3D) ultrasound is following the natural development of the imaging technology. This review of the technical applications and clinical aspects of the three-dimensional ultrasound is focused on vizualiztion of the fetal anatomy and the possibilities of this new technology and to increase awareness of its present clinical usefulness. Consulting specialists understand fetal pathology better and can better plan postnatal interventions. 3D ultrasound is a promising imaging method to image the fetus. Here are presented the methods for visualization in obstetrics, and the place of the ultrasound imaging in prenatal diagnosis. The role and value of this method will be in the focus of further studies.