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Introduction. In the treatment of stable angina in patients with concomitant bronchial asthma (BA), the bronchopulmonary system may be adversely affected by a number of drugs. Diltiazem may be the drug of choice when antianginal therapy with a pulseslowing effect is required, but β-blockers are contraindicated.Aim of the study. To compare the antianginal, pulse-slowing and vasoprotective effects of the calcium antagonist (CA) diltiazem, the calcium antagonist verapamil and the cardioselective beta-adrenoblocker (BAB) bisoprolol in the treatment of stable angina (SA) patients with concomitant BA.Materials and methods. The study included 60 patients with stable angina II-III FC with concomitant mild or moderate persistent BA. The patients were divided into three groups of 20 patients depending on antianginal drugs: Group 1 received betaadrenoblocker bisoprolol, Group 2 – calcium antagonist verapamil, Group 3 – calcium antagonist diltiazem. All patients underwent cardiography (Doppler EchoCG), external respiration function (ERF), endothelium-dependent vasodilation (EDVD) study at baseline and after 2, 4, 6 weeks of treatment.Results. No negative dynamics of FEV1 was found in patients of all groups during the study of ERF after 6 weeks of treatment. There was a statistically significant increase in FEV1 after 6 weeks of treatment in group 3 patients receiving diltiazem (p = 0.032). There was a statistically significant decrease in HR in all three groups during treatment, but in group 2 the dynamics were significantly lower when treated with verapamil. Analysis of mPAP of patients showed that it decreased in all groups after 6 weeks of treatment, but significant dynamics was noted only in Group 3. EDVD test after 6 weeks of treatment revealed positive dynamics, with a statistically significant increase in the index registered in patients of groups 1 and 3.Conclusions. In treatment of patients with stable angina with mild and intermediate persistent disease without exacerbation, antianginal therapy with the calcium antagonist diltiazem has marked antianginal, pulse-slowing action, has vasoprotective effect on the small and large circulatory vessels, improves the parameters of bronchial permeability.

Control over the strength of excitonic coupling in molecular dye aggregates is a substantial factor for the development of technologies such as light harvesting, optoelectronics, and quantum computing. According to the molecular exciton model, the strength of excitonic coupling is inversely proportional to the distance between dyes. Covalent DNA templating was proved to be a versatile tool to control dye spacing on a subnanometer scale. To further expand our ability to control photophysical properties of excitons, here, we investigated the influence of dye hydrophobicity on the strength of excitonic coupling in squaraine aggregates covalently templated by DNA Holliday Junction (DNA HJ). Indolenine squaraines were chosen for their excellent spectral properties, stability, and diversity of chemical modifications. Six squaraines of varying hydrophobicity from highly hydrophobic to highly hydrophilic were assembled in two dimer configurations and a tetramer. In general, the examined squaraines demonstrated a propensity toward face-to-face aggregation behavior observed via steady-state absorption, fluorescence, and circular dichroism spectroscopies. Modeling based on the Kühn-Renger-May approach quantified the strength of excitonic coupling in the squaraine aggregates. The strength of excitonic coupling strongly correlated with squaraine hydrophobic region. Dimer aggregates of dichloroindolenine squaraine were found to exhibit the strongest coupling strength of 132 meV (1065 cm

Nest aggregations of mud dauber wasps increase substrate heterogeneity and provide suitable sites for colonization by other invertebrate species. The mud dauber wasp Sceliphron fuscum Klug, 1801 (Hymenoptera: Sphecidae) abundantly occurs throughout the Seychelles Archipelago, Republic of Seychelles. Here, we estimated the taxonomic richness of the hymenopteran assemblage associated with S. fuscum’s nest aggregations, using material collected from the Inner Seychelles in 2016. Furthermore, we examine available historical survey data in order to assess possible changes in this association over decades. We discovered that from 1936 to 1938, seven hymenopteran species were associated with the nest aggregations of S. fuscum on Mahé and Praslin islands, representing six native taxa and one invasive species. From the material collected in 2016, we found one native and three invasive hymenopteran species associated to S. fuscum nests. Our findings could indicate a replacement of native species associated with the mud dauber wasps’ nest aggregations by recently introduced alien taxa on the Seychelles Archipelago.

Whilst the Arctic and boreal bumblebee fauna is increasingly studied worldwide, information is missing about the genetic connections between circum-boreal populations of some widespread species, especially those living in remote regions like North-East Siberia and Alaska. Here, we study one of the most common boreal bumblebee species, Bombus (Pyrobombus) jonellus (Kirby, 1802), as a model to investigate current circum-boreal genetic connections and relations with relictual populations in its post-glacial refugia in Southern Europe. Our haplotype network analysis based on a fragment of the cytochrome oxidase 1 gene reveals two main conspecific lineages, one in Europe including the Southern relictual populations from the Pyrenees and the second comprising Eastern Palearctic and Nearctic populations. However, West-Siberian populations of Bombus jonellus share haplotypes with the two distinct lineages. These results could indicate a postglacial, multidirectional and circum-boreal recolonization both in Europe and East-Palearctic from refugia in Siberia, in addition to other recolonization ways from Southern European refugia and Beringia. These findings highlight that a priori distant and isolated conspecific populations of B. jonellus could presently remain connected or have only presented a recent break in gene flow.

Bombus glacialis Friese, 1902 is an Arctic bumblebee that was thought to have a disjunctive range on the Arctic Ocean Islands, i.e., on Novaya Zemlya and Wrangel Island, with a 3600 km gap between these isolates. While the species status of the Novaya Zemlya’s population was recently confirmed using a molecular approach, its close affinities to the Wrangel’s isolate were proposed by a morphological similarity alone. Here, we report that B. glacialis samples from Wrangel share three unique COI haplotypes, which are closely related to each other and to those inferred from samples from Novaya Zemlya. Based on a combination of diagnostic morphological and molecular characters, the Wrangel’s population is described here as a new subspecies, Bombus glacialis marinae ssp. nov. Our novel data indicate that this species can be considered a polar relict that had had a much broader distribution along the continental margin of Eurasia during the Pleistocene. The two subspecies of B. glacialis were likely separated in the Middle Pleistocene (mean age = 0.27 Myr, 95% CI = 0.12–0.46 Myr) due to increasing of the sea level during an interglacial. The Novaya Zemlya Archipelago supported a B. glacialis population in the Late Pleistocene. Hence, this insular land was not covered completely by ice sheet during the Last Glacial Maximum. Our findings highlight that relict cold-adapted Pleistocene lineages still survive on remote islands in the Arctic Ocean and that such lineages are highly threatened by recent climate warming.

The focus of this study is to summarize the data on the distribution and foraging preference of Bombus (Megabombus) consobrinus Dahlbom, 1832 in the European North of Russia. The range of B. consobrinus in this region mostly repeats the disjunctions of the range of Aconitum septentrionale that is also known in Scandinavia. In other regions of Northern Eurasia, the close relationship of B. consobrinus with Aconitum is not obvious. This bumblebee species may be regarded as oligolectic in Northern Europe and the European North of Russia. We assume the presence of a coadaptive relationship of this bumblebee species with A. septentrionale in this region that presumably have been caused by the complex history of B. consobrinus in the European North.

Recent multi-locus phylogenetic studies repeatedly showed that what was thought to be the Clouded Apollo butterfly Parnassius mnemosyne (Linnaeus, 1758) represents a cryptic species complex. This complex contains at least three distant species-level phylogenetic lineages. Here, we compile a set of morphology- and DNA-based evidences supporting the distinctiveness of two species in this group, i.e. P. mnemosyne s. str. and P. nebrodensis Turati, 1907 stat. rev. These species can be distinguished from each other based on a combination of diagnostic characters in the male genitalia structure, wing scale patterns, and the forewing venation. The species status of P. nebrodensis is supported based on unique nucleotide substitutions in the mitochondrial (COI, ND1, and ND5) and nuclear (Wg and EF-1a) genes. P. nebrodensis is endemic to the Western Mediterranean Region. This species shares a disjunctive range through the Pyrenees, Western and Central Alps, Apennines, and the Nebrodi and Madonie mountains on Sicily. Altogether 38 nominal taxa initially described as P. mnemosyne subspecies are considered here to be junior synonyms of P. nebrodensis. At first glance, P. nebrodensis can be assessed as an endangered species due to its restricted distribution, narrow range of habitats, and ongoing population decline. Isolated populations of this species scattered through mountain ranges need special management and conservation efforts.

This study (1) displays markings pattern of male and female specimens of Arctia tundrana from various parts of its broad range; (2) illustrates a paratype male specimen of this species with its genitalia and aedeagus; (3) presents a few additional occurrences of A. tundrana supplementing the data set published in our earlier paper (Bolotov et al. 2015); (4) provides an updated map of the species’ occurrences; and (5) discusses its imaginal phenology based on long-term occurrence data.

In this study, we examine a recent sample containing two hawk moth species (Lepidoptera: Sphingidae) from Praslin and Mahé, Seychelles. It was widely accepted that the hawk moth species Temnora peckoveri (Butler, 1876) has a disjunctive range covering Madagascar and the Inner Seychelles. However, the Seychelles population of what was thought to be Temnora peckoveri shares a set of diagnostic morphological differences from both this species and T. fumosa (Walker, 1856) in the male genitalia structure. In particular, it remotely resembles Temnora fumosa based on the structure of aedeagus but differs from T. fumosa and T. peckoveri with respect to the harpe shape. Based on this evidence, we describe Temnora fumosa seychellensis Bolotov & Spitsyn ssp. nov. as a subspecies with restricted range, being endemic to the Inner Seychelles. The range of this subspecies covers Praslin (first record), Mahé, Silhouette, La Digue, Cousine, and Denis. Additionally, we report on recent occurrences of Agrius convolvuli (Linnaeus, 1758) from Praslin (second record) and Mahé. The adult moths were recorded feeding on inflorescences of Dracaena reflexa var. angustifolia Baker (Asparagaceae) for the first time. Finally, we present a complete list of Temnora fumosa seychellensis ssp. nov. and Agrius convolvuli occurrences from Seychelles.

The quasi-potential approach is very famous in modern relativistic particles physics. This approach is based on the so-called covariant single-time formulation of quantum field theory in which the dynamics of fields and particles is described on a space-like three-dimensional hypersurface in the Minkowski space. Special attention in this approach is paid to methods for constructing various quasi-potentials. The quasipotentials allow to describe the characteristics of relativistic particles interactions in quark models such as amplitudes of hadron elastic scatterings, mass spectra, widths of meson decays and cross sections of deep inelastic scatterings of leptons on hadrons. In this paper SturmLiouville problems with periodic boundary conditions on a segment and a positive half-line for the 2m-order truncated relativistic finite-difference Schrdinger equation (LogunovTavkhelidzeKadyshevsky equation, LTKT-equation) with a small parameter are considered. A method for constructing of asymptotic eigenfunctions and eigenvalues in the form of asymptotic series for singularly perturbed SturmLiouville problems with periodic boundary conditions is proposed. It is assumed that eigenfunctions have regular and boundary-layer components. This method is a generalization of asymptotic methods that were proposed in the works of A. N. Tikhonov, A. B. Vasilyeva, and V. F Butuzov. We present proof of theorems that can be used to evaluate the asymptotic convergence for singularly perturbed problems solutions to solutions of degenerate problems when 0 and the asymptotic convergence of truncation equation solutions in the case m. In addition, the SturmLiouville problem on the positive half-line with a periodic boundary conditions for the quantum harmonic oscillator is considered. Eigenfunctions and eigenvalues are constructed for this problem as asymptotic solutions for 4-order LTKT-equation.

Geothermal springs are known to harbor unusual assemblages of dragonflies and damselflies worldwide. A review of original records and the body of available literature revealed that 27 Odonata species were recorded from hot springs in Russia so far and that the successful larval development in geothermal environments was discovered for 17 species. Among them, four species exclusively inhabit hot springs, i.e. Mnais costalis, Anotogaster sieboldii, Orthetrum melania (Kunashir Island), and O. albistylum (Eastern Siberia). In Russia, these southern species are unable to develop beyond geothermally heated water bodies due to cold climate, and they exist as local geothermal populations there. Here, we report on several novel records of Odonata species from geothermal springs in eastern Russia. Four species were recorded on the Kunashir Island (Kurile Archipelago): Mnais costalis (adults), Anotogaster sieboldii (adults and larvae), Orthetrum melania (adults), and Sympetrum pedemontanum elatum (adults and larvae). Two species were found in the Kamchatka Peninsula, i.e. Libellula quadrimaculata (freshly emerged imago and exuvia) and Aeshna juncea (larvae). To explain the origin of isolated geothermal populations of Odonata, three alternative hypotheses can be proposed as follows: (1) pre-glacial relicts; (2) mid-Holocene relicts (since the Holocene Climate Optimum); and (3) recent (late Holocene) populations founded by long-distance dispersal events. These scenarios are yet to be tested by means of a molecular approach.

It is known, that the application of Forster Resonance Energy Transfer (FRET) between two dyes absorbing in different spectrum regions allows shifting of the excitation wavelength into the blue region. We investigated spectral properties and FRET between styryl (St) and squaraine (Sq) dyes to evaluate the possibility of using such FRET-pair to determine conformational changes in proteins. Spectral properties of St and Sq in polar (phosphate buffer), nonpolar (chloroform) solvents and in non-covalent complexes with BSA (bovine serum albumin) and HSA (human serum albumin) were studied. The effect of changes in protein conformation induced by urea solutions of various concentrations on the spectral properties of dyes and energy transfer efficiency was also studied. It was found that the polarity of the medium affects the fluorescent properties of dyes in different ways: styryl dye has a positive solvatofluorochromism, while the squaraine dye has the negative one. Both dyes show negative solvatochromism, and their quantum yields substantially increase in a less polar environment. Despite the spectral differences of the St-BSA and St-HSA complexes, their quantum yields are the same, and in case of squaraine dye the quantum yield for Sq-HSA is almost twice higher that for Sq-BSA. When the conformation of protein changes, the dyes go into the aqueous phase, as a result the intensity of their fluorescence decreases almost to the level of fluorescence of the free dye in phosphate buffer. There are two bands corresponding to the donor and acceptor in the fluorescence spectra of the double complexes St-BSA-Sq and St-HSA-Sq. The pair of styryl-squaraine dyes is differently sensitive to the conformation changes in these albumins caused by urea: the FRET efficiency for HSA changes by 11.5 times, while for BSA only by 2.1 times. Thus, the St–Sq pair has the ability to detect a change in the conformation of proteins, but the efficiency of the pair depends on the properties of the protein under study.

In this correspondence, we report on the field observations of puddling by butterflies on warm mineral water from hot springs in Northern Asia.

Papilio demoleus Linnaeus, 1758 (Lepidoptera: Papilionidae) is a widespread pest of citrus plants originating from the Oriental Region. Here, we report on the discovery of its non-native population recently established on Mahé, Seychelles. Papilio demoleus was probably accidentally introduced to Mahé a few years ago (first records in November 2016). Morphologically, the Seychelles population is identified as the subspecies Papilio demoleus malayanus Wallace, 1865 from Southeast Asia. It was one of the most abundant butterfly species at the southeastern edge of Mahé in January 2020. Our finding highlights that generalist butterfly species associated with agricultural plants are rapidly spreading throughout remote oceanic islands such as the Inner Seychelles.

В статье отражена проблема обучения детей грамоте и предложено пособие для ее решения, в котором приведена динамика развития детей после коррекционной работы с данным пособием.

The name nebrodensis Turati is shown to be unavailable and the first available name for the species is chosen by invoking the First Reviser Principle under the ICZN Code as Parnassius turatii Fruhstorfer, 1908. Rothschild was determined to be the first author to confer availability on the name nebrodensis by application of the Code.

An advanced thermoelectric system has been developed to set a low temperature to the heat-generating power semiconductor devices. The system provides the setting the temperature down to -60°C in a volume of 80x40x12 mm3 at the heat-release up to 13 W. Accuracy of setting the temperature is in the range of ± 3°C. The design of developed system is adopted for long-term tests, including tests in harsh environment. The cooling chamber in may be placed up to 6 m away from the heat exchanger and control unit. One of the main features of new development is its mobility and easy installation when carrying out various types of tests.

Global environmental changes may cause dramatic insect declines but over century-long time series of certain species' records are rarely available for scientific research. The Menetries' Tiger Moth (Arctia menetriesii) appears to be the most enigmatic example among boreal insects. Although it occurs throughout the entire Eurasian taiga biome, it is so rare that less than 100 specimens were recorded since its original description in 1846. Here, we present the database, which contains nearly all available information on the species' records collected from 1840s to 2020. The data on A. menetriesii records (N = 78) through geographic regions, environments, and different timeframes are compiled and unified. The database may serve as the basis for a wide array of future research such as the distribution modeling and predictions of range shifts under climate changes. It represents a unique example of a more than century-long dataset of distributional, ecological, and phenological data designed for an exceptionally rare but widespread boreal insect, which primarily occurs in hard-to-reach, uninhabited areas of Eurasia.

Bolotov, Ivan N., Kolosova, Yulia S., Chapurina, Yulia E., Spitsyna, Elizaveta A., Spitsyn, Vitaly M. (2021): A new genus and species of planthopper from Seychelles endemic palm forest (Hemiptera: Fulgoromorpha: Derbidae). Journal of Natural History 55 (19-20): 1311-1321, DOI: 10.1080/00222933.2021.1947536, URL: http://dx.doi.org/10.1080/00222933.2021.1947536

A bumblebee community was studied at Pymvashor, the only thermal spring in North European Russia. The bumblebee fauna comprised 12 species, which is a large number when compared to other native tundra ecosystems. Most of the species recorded were ubiquitous, 3 were forest species and 2 were typical tundra species. The presence of the ubiquitous and forest species in the bumblebee community appears to be due to the landscape features and the perennial impact of the hot springs, under the influence of which specific extrazonal ecosystems arise that are different from those typical of the tundra zone.

Novel indolenine based norsquaraine dyes, wherein the oxygen of the squaric acid bridge was substituted with a barbituric or a dicyanomethylene group, were synthesized and their molecular structure, spectral and luminescent properties were compared to those of analogous squaraine dyes. The molecular structure was investigated using X-ray analysis, NMR spectroscopy and ab initio DFT B3LYP/6-311G (d, p) simulations. The calculated populations of possible conformers and the barriers of internal rotations were found to be in good agreement with the experimental data. Norsquaraines absorb and emitt light within the same long-wavelength spectral range as the corresponding squaraines but due to intramolecular H-bonds and increased conformational rigidity they were less sensitive to solvent polarity and the presence of protein (BSA).

Aim To compare efficacy and safety of treatments with the calcium antagonist (CA) verapamil, the cardioselective β-blocker (BB) bisoprolol, and a combination therapy with bisoprolol and amlodipine in patients with stable angina (SA) with concurrent mild and moderate, persistent bronchial asthma (BA). Material and methods This open, prospective, randomized, comparative study included 120 patients with an IHD+BA comorbidity. Of these patients, 60 had mild persistent BA and 60 had moderate persistent BA. Each group was divided into 3 subgroup, each including 20 patients, based on the used regimen of antianginal therapy. Stepwise dose titration was performed every 2 weeks (subgroup 1 received the BB bisoprolol 2.5 mg – 5 mg – 10 mg; subgroup 2 received the CA verapamil 240 mg – 240 mg – 240 mg; subgroup 3 received bisoprolol 2.5 mg followed by the combination treatment with bisoprolol and amlodipine as a fixed combination 5+5 mg). All patients underwent a complete clinical and instrumental examination at baseline and at 2, 4, and 6 weeks of treatment. The antianginal effectivity and the effect on bronchial patency were evaluated. Results In patients with SA and mild persistent BA, the study of external respiration function (ERF) at 2, 4, and 6 weeks of treatment did not detect any significant difference in the forced expiratory volume in 1 second (FEV1) between the treatment subgroups. In patients with SA and moderate persistent BA receiving the treatment, a significant decrease in FEV1 (р=0.022) was observed in subgroup 1 receiving bisoprolol 10 mg at 6 weeks of treatment. In subgroups 2 and 3 during the treatment, significant differences were absent. In patients with SA and mild or moderate persistent BA, the heart rate was significantly decreased in all three subgroups; however, in subgroup 2 receiving verapamil, the changes were considerably smaller than in other subgroups.Conclusion The study results showed that the BB bisoprolol with dose titration every two weeks from 2.5 to 10 mg or the combination treatment with the BB bisoprolol and the CA amlodipine can be used as the antianginal therapy in patients with SA and mild persistent BA. The BB bisoprolol may be used in patients with SA and moderate persistent BA as the antianginal therapy, but only at doses not exceeding 5 mg to avoid the development of bronchial obstruction. The combination therapy with the BB bisoprolol 5 mg and the CA amlodipine 5 mg is indicated to enhance antianginal and vasoprotective effects.

A description of the system for setting a lowered environment temperature based on Peltier modules is presented. The development was carried out taking into account the peculiarities of the radiation test: minimum distance from exposure source to DUT; stand-alone use during long-term radiation test; minimum overall dimensions, transportability and flexibility when conducting tests on various exposure sources. Cooling box consists of 3-level stacked assembly of the Peltier modules, which is cooled by liquid refrigerant. Control of the assembly of Peltier modules was performed according to proportional integral and differential algorithm, provided by microcontroller with feedbacks on Pt thermoresistors. The system provides temperature stabilization of the DUT up to 23х17х12 mm in size and with released heat power up to 0.4 W at minus (60 ± 3) oС. Temperature establishing time is less than 10 minutes. Typical power consumption of developed system is about 500W.

Dye molecules that absorb light in the visible region are key components in many applications, including organic photovoltaics, biological fluorescent labeling, super-resolution microscopy, and energy transport. One family of dyes, known as squaraines, has received considerable attention recently due to their favorable electronic and photophysical properties. In addition, these dyes have a strong propensity for aggregation, which results in emergent materials properties, such as exciton delocalization. This will be of benefit in charge separation and energy transport along with fundamental studies in quantum information. Given the high structural tunability of squaraine dyes, it is possible that exciton delocalization could be tailored by modifying the substituents attached to the π-conjugated network. To date, limited theoretical studies have explored the role of substituent effects on the electronic and photophysical properties of squaraines in the context of DNA-templated dye aggregates and resultant excitonic behavior. We used ab initio theoretical methods to determine the effects of substituents on the electronic and photophysical properties for a series of nine different squaraine dyes. Solvation free energy was also investigated as an insight into changes in hydrophobic behavior from substituents. The role of molecular symmetry on these properties was also explored via conformation and substitution. We found that substituent effects are correlated with the empirical Hammett constant, which demonstrates their electron donating or electron withdrawing strength. Electron withdrawing groups were found to impact solvation free energy, transition dipole moment, static dipole difference, and absorbance more than electron donating groups. All substituents showed a redshift in absorption for the squaraine dye. In addition, solvation free energy increases with Hammett constant. This work represents a first step toward establishing design rules for dyes with desired properties for excitonic applications.

Although the Odonata are common inhabitants of various extreme environments such as geothermal springs, brackish wetlands, mangroves, and volcanic lakes, the assemblages of this group associated with extreme habitats in Australasia are rather poorly known. Here, we combine museum collection data and published reports on Odonata from extreme habitats on Flores Island, Lesser Sunda Archipelago. The highly acidic Sano Nggoang Crater Lake (mean pH = 3.17) on Flores houses seven species as follows: Agriocnemis pygmaea, Xiphiagrion cyanomelas (Coenagrionidae), Neurothemis ramburii, Orthetrum pruinosum pruinosum, O. sabina, O. testaceum soembanum (Libellulidae), and Anax gibbosulus (Aeshnidae). A coastal marsh site with slightly brackish water on Flores harbors at least five dragonfly species as follows: Diplacodes trivialis, Neurothemis intermedia excelsa, N. terminata, Pantala flavescens, and Rhyothemis phyllis ixias (Libellulidae). The migratory dragonfly Pantala flavescens was a single species recorded on the waterless Kanawa Island near the western edge of Flores. Our findings suggest that extreme habitats in eastern Indonesia primarily colonized by widespread generalist Odonata species. Finally, an updated checklist of Odonata species recorded from Flores Island was compiled. Our survey of museum specimens recovered two species not found on existing species lists for Flores: Neurothemis intermedia excelsa and Pantala flavescens.

Freshwater mussels (Bivalvia: Unionoida) play an important role in freshwater habitats as ecosystem engineers of the water environment. Duck mussel Anodonta anatina is widely distributed throughout Europe, Siberia, and Western and Central Asia, which makes it a convenient object for biogeographic studies. In this study, we analyzed the divergence of A. anatina populations and discovered a separate genetic lineage distributed in rivers of the Azov Sea basin. This was confirmed by the high genetic distances between this group and previously defined populations, and by the position of this clade in the Bayesian phylogeny calibrated by an external substitution rate. Based on our approximate Bayesian computation (ABC) analysis, biogeographic scenarios of A. anatina dispersal in Europe and Northern, Western, and Central Asia over the Neogene&ndash, Quaternary were simulated. The haplogroup&rsquo, s isolation in the rivers of the Azov Sea basin most likely occurred in the Late Pliocene that was probably facilitated by rearrangement of freshwater basins boundaries in the Ponto-Caspian Region. Population genetic indices show the stability of this group, which allowed it to exist in the river basins of the region for a long time. The discovery of a long-term refugium in the rivers of the Azov Sea led to a better understanding of freshwater fauna evolution in the Neogene&ndash, Quaternary and highlighted the importance of conservation of these freshwater animals in the region as a source of unique genetic diversity.

The work was to develop a highly sensitive, fluorescence based method to monitor conformational changes in proteins. With this aim, the influence of unfolding of bovine serum albumin (BSA) molecules on the spectral properties of squaraine dye Sq1 and dicyanomethylene squaraine dye Sq2 was investigated. The conformational changes were initiated by urea in the concentration of up to 8 M. The following three intensity-based methods of using these dyes were evaluated: (i) dye Sq1 was covalently bound to BSA; (ii) dye Sq1 was noncovalently bound to BSA; and (iii) dye Sq2 was noncovalently bound to BSA. The change in the fluorescence intensity of these dyes upon the BSA unfolding was measured. An additional, fourth method was based on the measurements of the change in the Förster Resonance Energy Transfer (FRET) between Sq1 (donor) covalently bound to BSA and Sq2 (acceptor) able to noncovalent binding with BSA. The change in the FRET was due to the BSA unfolding initiated by urea. The absorption and emission maxima measured in water were 635 / 645 nm for dye Sq1 and 654 / 670 nm for Sq2. Addition of 7–8 M urea to aqueous solutions of dyes Sq1 and Sq2 increased their fluorescence intensities by factor of 1.5 and 2.3 while the quantum yield increased by factor of 1.8 and 1.9, respectively. The absorption intensity of Sq1 in the presence of urea had a tendency to decrease while for Sq2 it increased by factor of 1.3. The formation of complexes of dyes Sq1 and Sq2 with BSA as well as the covalent conjugation of Sq1 with BSA caused the red-shift in the absorption and emission maxima and an increase in the quantum yields. Unfolding of BSA molecule resulted in a blue-shift of the absorption and emission bands and a 3-fold decrease in the fluorescence intensity of Sq2–BSA complex. Unfolding of protein in Sq1–BSA conjugate had almost no effect on the position and shape of the absorption and emission bands but decreased the fluorescence intensity by about 20%. At the same time, according to the fourth method the efficiency of FRET from Sq1 conjugated with BSA, to non-conjugated Sq2 exhibited much more pronounced decrease (~10 times) upon BSA unfolding. Thus, the FRET based approach was found to be more sensitive compared to the intensity based methods and therefore can changes in BSA.