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PurposeThe purpose of this study was to evaluate the epidemiology, etiology, clinical assessment, investigation, management, and visual consequences of high myopia (≤-6 diopters [D]) in infants and young children.FindingsHigh myopia is rare in pre-school children with a prevalence less than 1%. The etiology of myopia in such children is different than in older children, with a high rate of secondary myopia associated with prematurity or genetic causes. The priority following the diagnosis of high myopia in childhood is to determine whether there is an associated medical diagnosis that may be of greater overall importance to the health of the child through a clinical evaluation that targets the commonest features associated with syndromic forms of myopia. Biometric evaluation (including axial length and corneal curvature) is important to distinguishing axial myopia from refractive myopia associated with abnormal development of the anterior segment. Additional investigation includes ocular imaging, electrophysiological tests, genetic testing, and involvement of pediatricians and clinical geneticists is often warranted. Following investigation, optical correction is essential, but this may be more challenging and complex than in older children. Application of myopia control interventions in this group of children requires a case-by-case approach due to the lack of evidence of efficacy and clinical heterogeneity of high myopia in young children.ConclusionsHigh myopia in infants and young children is a rare condition with a different pattern of etiology to that seen in older children. The clinical management of such children, in terms of investigation, optical correction, and use of myopia control treatments, is a complex and often multidisciplinary process.

The crisis of moral principles of social development at the present stage, the need of Russian law to create new legal traditions, the need to form a new scale of social values necessitate the assessment of the existing criminal procedure regulation for compliance with moral and ethical standards, the main axiological determinants of modern society, among which faith and trust occupy a special place. Through these categories, it is necessary to assess the understanding of evidence reliability in criminal proceedings. Purpose: to analyze the evidence reliability by interpreting its criteria in law enforcement practice. Methods: methods of comparative and comparative law research, description, interpretation; axiological and axiomatic methods; specific scientific methods: legal-dogmatic and interpretation of legal norms. Results: the study allows to conclude that it is necessary to assess the evidence reliability due not only to its procedural significance, but also to its moral essence. The analysis of judicial practice allows to argue the thesis that, unlike faith, which does not require proof, reliability needs confirmation, but in practice the criteria for assessing the evidence reliability are highly differentiated.

The article examines fraudulent discourse – a special type of discourse, the purpose of which is to take possession of other people’s property and obtain benefits through deceit or breach of trust. The author differentiates the legal understanding of the phenomenon of “fraud” and its perception in the ordinary worldview. It is proved that the concept of fraudulent discourse should include a number of communicative acts qualified by the Criminal Code of the Russian Federation as theft or extortion. The central genre of fraudulent discourse is the speech genre “swindle” / “con game”, based on the conscious and purposeful manipulative influence of the initiator of communication (fraudster) on the addressee (victim). Nowadays, the wide spread of remote communication determines the predominance of “swindle” using various technical means that provide synchronous or asynchronous interaction between the subjects of communication. The “swindle” / “con game” genre embodies the formulaicity of fraudulent discourse and is implemented through a limited number of strategies, each of which uses a stable set of means of influence. The main strategies are the appeal to fear, the appeal to luck and creation of the illusion of routine. As part of the fear appeal strategy, three tactics are distinguished, namely, the tactics of instilling fear for one’s prosperity and well-being, the tactics of instilling fear for the safety of a family member, and the tactics of instilling fear of public reprimand. The analysis is carried out by a continuous sampling method and is based on messages (more than 400 units) of potential victims or victims of technically mediated fraud, presented in the form of a narrative and posted on various communication resources in the media space.

The structure of lactose permease, stabilized in a periplasmic open conformation by two Gly to Trp replacements (LacYww) and complexed with a nanobody directed against this conformation, provides the highest resolution structure of the symporter. The nanobody binds in a different manner than two other nanobodies made against the same mutant, which also bind to the same general region on the periplasmic side. This region of the protein may represent an immune hotspot. The CDR3 loop of the nanobody is held by hydrogen bonds in a conformation that partially blocks access to the substrate-binding site. As a result, kon and koff for galactoside binding to either LacY or the double mutant complexed with the nanobody are lower than for the other two LacY/nanobody complexes though the Kd values are similar, reflecting the fact that the nanobodies rigidify structures along the pathway. While the wild-type LacY/nanobody complex clearly stabilizes a similar 'extracellular open' conformation in solution, judged by binding kinetics, the complex with wild-type LacY did not yet crystallize, suggesting the nanobody does not bind strongly enough to shift the equilibrium to stabilize a periplasmic side-open conformation suitable for crystallization. However, the similarity of the galactoside binding kinetics for the nanobody-bound complexes with wild type LacY and with LacYWW indicates that they have similar structures, showing that the reported co-structures reliably show nanobody interactions with LacY.

The lactose permease of Escherichia coli (LacY), a dynamic polytopic membrane transport protein, catalyzes galactoside/H+ symport and operates by an alternating access mechanism that exhibits multiple conformations, the distribution of which is altered by sugar-binding. Camelid nanobodies were made against a double-mutant Gly46 → Trp/Gly262 → Trp (LacYWW) that produces an outward-open conformation, as opposed to the cytoplasmic open-state crystal structure of WT LacY. Nanobody 9047 (Nb9047) stabilizes WT LacY in a periplasmic-open conformation. Here, we describe the X-ray crystal structure of a complex between LacYWW, the high-affinity substrate analog 4-nitrophenyl-α-d-galactoside (NPG), and Nb9047 at 3-Å resolution. The present crystal structure demonstrates that Nb9047 binds to the periplasmic face of LacY, primarily to the C-terminal six-helical bundle, while a flexible loop of the Nb forms a bridge between the N- and C-terminal halves of LacY across the periplasmic vestibule. The bound Nb partially covers the vestibule, yet does not affect the on-rates or off-rates for the substrate binding to LacYWW, which implicates dynamic flexibility of the Nb-LacYWW complex. Nb9047-binding neither changes the overall structure of LacYWW with bound NPG, nor the positions of side chains comprising the galactoside-binding site. The current NPG-bound structure exhibits a more occluded periplasmic vestibule than seen in a previous structure of a (different Nb) apo-LacYWW/Nb9039 complex that we argue is caused by sugar-binding, with major differences located at the periplasmic ends of transmembrane helices in the N-terminal half of LacY.

Over the last five years, inorganic lead halide perovskite nanowires have emerged as prospective candidates to supersede standard semiconductor analogs in advanced photonic designs and optoelectronic devices. In particular, CsPbX3 (X = Cl, Br, I) perovskite materials have great advantages over conventional semiconductors such as defect tolerance, highly efficient luminescence, and the ability to form regularly shaped nano- and microcavities from solution via fast crystallization. However, on the way of electrically pumped lasing, the perovskite nanowires grown on transparent conductive substrates usually suffer from strong undesirable light leakage increasing their threshold of lasing. Here, we report on the integration of CsPbBr3 nanowires with nanostructured indium tin oxide substrates possessing near-unity effective refractive index and high conductivity by using a simple wet chemical approach. Surface passivation of the substrates is found out to govern the regularity of the perovskite resonators’ shape. The nanowires show room-temperature lasing with ultrahigh quality factors (up to 7860) which are up to four times higher than that of similar structures on a flat indium tin oxide layer, resulting in more than twofold reduction of the lasing threshold for the nanostructured substrate. Numerical modeling of eigenmodes of the nanowires confirms the key role of low-refractive-index substrate for improved light confinement in the Fabry–Pérot cavity which results in superior laser performance.

The paper considers four methods of drying kelp (Laminaria digitata): convective dehydration at an air temperature of minus 5 oС, relative air humidity of 55 and 90 %; vacuum drying at a metal carrier temperature of 20°C and a vacuum depth of 250 Pa; convective dehydration at an air temperature of 20°C and a relative air humidity of 35 %. In dried algae, the final moisture content, water activity, and strength characteristics were evaluated. To determine the swelling coefficient, dry algae were pre-crushed. It was established that at a temperature of minus 5°C and a relative air humidity of 90 %, dehydration had an unsatisfactory result in terms of the final moisture content of the finished product 27.4 %. This type of drying at a relative air humidity of 55 % provides a final moisture content of the product (12.3 ± 0.48) % and a relatively high swelling coefficient of 6.9. Algae, dehydrated at minus 5°C, had elastic properties, in contrast to other considered methods. Vacuum drying made it possible to obtain the minimum final moisture content of dry algae and the highest swelling coefficient of 7.7. However, drying to constant weight lasted 25.5 hours. Of the considered drying options, the convective method (t = 20°C) showed the highest rate. The duration of drying to constant weight was 8 hours. The final moisture content of the product was acceptable (9.8±1.52) %. However, the water activity after algae swelling had the highest value of 0.97±0.004, and the swelling coefficient had a relatively low value of 6.5. In further studies, it is planned to study the effect of various methods of algae dehydration on the preservation of its biologically active substances.

In different regions of Mexico, speech may have local distinctive characteristics at different language levels. The article deals with the grammatical features of Spanish in the territory of the Mexican state of Oaxaca. The article raises the following questions: 1) whether Spanish in the territory of the Mexican state of Oaxaca has certain local characteristics; 2) whether there is a formation of local territorial norms in this state or we can only talk about the occasional use of certain grammatical forms. Examples are given showing the presence of grammatical features in the structure of the Spanish language in the state of Oaxaca. A comparison is made between Oaxacan and the Mexican national version of Spanish. The scientific novelty of the work lies in the lack of work on this issue in domestic and foreign Spanish studies. The analysis of grammar discrepancies was carried out on the basis of a survey and questioning of residents of the capital and state of Oaxaca, as well as a study of scientific literature and Internet resources. The results of the research may be used in future studies of Spanish.

The lactose permease of Escherichia coli (LacY), a highly dynamic polytopic membrane protein, catalyzes stoichiometric galactoside/H(+) symport by an alternating access mechanism and exhibits multiple conformations, the distribution of which is altered by sugar binding. We have developed single-domain camelid nanobodies (Nbs) against a LacY mutant in an outward (periplasmic)-open conformation to stabilize this state of the WT protein. Twelve purified Nbs inhibit lactose transport in right-side-out membrane vesicles, indicating that the Nbs recognize epitopes on the periplasmic side of LacY. Stopped-flow kinetics of sugar binding by WT LacY in detergent micelles or reconstituted into proteoliposomes reveals dramatic increases in galactoside-binding rates induced by interaction with the Nbs. Thus, WT LacY in complex with the great majority of the Nbs exhibits varied increases in access of sugar to the binding site with an increase in association rate constants (kon) of up to ∼ 50-fold (reaching 10(7) M(-1) ⋅ s(-1)). In contrast, with the double-Trp mutant, which is already open on the periplasmic side, the Nbs have little effect. The findings are clearly consistent with stabilization of WT conformers with an open periplasmic cavity. Remarkably, some Nbs drastically decrease the rate of dissociation of bound sugar leading to increased affinity (greater than 200-fold for lactose).

Background and Aim: Antibiotics’ resistance is the leading cause of complications in the treatment of urinary tract infections. This study aimed to screen the antimicrobial potential of 8 plants from Cameroon against multi-resistant uropathogenic (MRU) bacteria and to investigate their antibioresistance reversal properties. Method: Bioactive compounds were extracted from leaves of Leucanthemum vulgare, Cymbopogon citratus, Moringa oleifera and Vernonia amygdalina; barks of Cinchona officinalis and Enantia chlorantha barks and seeds of Garcinia lucida and leaves and seeds of Azadirachta indica using water and ethanol as solvents. The extracts were tested against Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 6538 and Candida albicans 10231 using the well diffusion and the broth microdilution methods. The antibiotic-resistance reversal activity was assessed against selected MRU bacteria. The phytochemical composition and the elemental composition of the most active extracts were assessed respectively using HPLC-MS/MS and X-ray fluorescence (XRF) spectrometry. Results: Among the most active plants, in decreasing order of antimicrobial activity we found ethanolic (EE) and aqueous extracts (AE) of E. chloranta bark (ECB), EE of L. vulgare leaves and G. lucida seeds. The best synergies between common antibiotics and extracts were found with EE-ECB which well-modulated kanamycin nitrofurantoin and ampicillin. All the compounds identified in EE-ECB were alkaloids and the major constituents were palmatine (51.63%), columbamine+7,8-dihydro-8-hydroxypalmatine (19.21%), jatrorrhizine (11.02%) and pseudocolumbamine (6.33%). Among the minerals found in EE-ECB (S, Si, Cl, K, Ca, Mn, Fe, Zn and Br), Br, Fe and Cl were the most abundant with mean fluorescence intensities of 4.6529, 3.4854 and 2.5942 cps/uA respectively. Conclusions: The ethanol extract of the bark of E. chlorantha has remarkable, broad-spectrum antimicrobial and contains several palmatine derivatives.

Galactoside/H(+) symport across the cytoplasmic membrane of Escherichia coli is catalyzed by lactose permease (LacY), which uses an alternating access mechanism with opening and closing of deep cavities on the periplasmic and cytoplasmic sides. In this study, conformational changes in LacY initiated by galactoside binding were monitored in real time by Trp quenching/unquenching of bimane, a small fluorophore covalently attached to the protein. Rates of change in bimane fluorescence on either side of LacY were measured by stopped flow with LacY in detergent or in proteoliposomes and were compared with rates of galactoside binding. With LacY in proteoliposomes, the periplasmic cavity is tightly sealed and the substrate-binding rate is limited by the rate of opening of this cavity. Rates of opening, measured as unquenching of bimane fluorescence, are 20-30 s(-1), independent of sugar concentration and essentially the same in detergent or in proteoliposomes. On the cytoplasmic side of LacY in proteoliposomes, slow bimane quenching (i.e., closing of the cavity) is observed at a rate that is also independent of sugar concentration and similar to the rate of sugar binding from the periplasmic side. Therefore, opening of the periplasmic cavity not only limits access of sugar to the binding site of LacY but also controls the rate of closing of the cytoplasmic cavity.

Here we describe the X-ray crystal structure of a double-Trp mutant (Gly46→Trp/Gly262→Trp) of the lactose permease of Escherichia coli (LacY) with a bound, high-affinity lactose analog. Although thought to be arrested in an open-outward conformation, the structure is almost occluded and is partially open to the periplasmic side; the cytoplasmic side is tightly sealed. Surprisingly, the opening on the periplasmic side is sufficiently narrow that sugar cannot get in or out of the binding site. Clearly defined density for a bound sugar is observed at the apex of the almost occluded cavity in the middle of the protein, and the side chains shown to ligate the galactopyranoside strongly confirm more than two decades of biochemical and spectroscopic findings. Comparison of the current structure with a previous structure of LacY with a covalently bound inactivator suggests that the galactopyranoside must be fully ligated to induce an occluded conformation. We conclude that protonated LacY binds D-galactopyranosides specifically, inducing an occluded state that can open to either side of the membrane.

Lactose permease of Escherichia coli (LacY) is highly dynamic, and sugar binding causes closing of a large inward-facing cavity with opening of a wide outward-facing hydrophilic cavity. Therefore, lactose/H+ symport via LacY very likely involves a global conformational change that allows alternating access of single sugar- and H+-binding sites to either side of the membrane. Here, in honor of Stephan H. White’s seventieth birthday, we review in camera the various biochemical/biophysical approaches that provide experimental evidence for the alternating access mechanism.

The practice of ensuring the right to protection of persons from suspicion or accusation in some countries, including taking into account the decisions of the European Court of Human Rights, indicates that not all issues of its legal regulation have been resolved to a degree that satisfies science and practice, and there are violations of human rights enshrined in international legal standards, the principle of adversarial parties in criminal proceedings is not fully implemented, so there is increasing scientific interest in problematic issues of protection from criminal prosecution. The purpose of the study is to analyze international standards in the field of human rights, as well as the experience of Russia and some foreign countries in the field of ensuring the right to protection from criminal prosecution in criminal proceedings, to identify problematic issues of a legal nature in this part and to formulate scientifically based recommendations for their solution (minimization). The methodological basis of the study is also the formallogical and comparative-legal methods, taking into account the processes of development of the regulatory framework.

Buildings are responsible for 40% of energy consumption annually in Europe, along with the respective greenhouse gas emissions. To mitigate these impacts, intensive research is ongoing in the sector of the Nearly Zero-Energy Buildings (NZEBs). However, as it is expected that the operational energy of future buildings becomes greener and more efficient, impacts related to the embodied energy of building materials becomes of more significance. Thus, choices on building materials are of crucial importance as they affect the energy performance of the building envelope and its environmental impacts. The objective of this study was to implement preliminary Life Cycle Assessment (LCA) on new advanced building materials, with the final scope to achieve lower embodied carbon in NZEBs. The materials examined are concretes and aerogels for wall façades. Design of sustainable advanced materials and building envelope components is expected to improve the overall energy performance of buildings, including NZEBs. The study findings provide clear evidence on the necessity for further research on the topic, as lack of embodied impacts’ data of novel materials is presented in literature and adds to the discussion around NZEBs.

Electrogenic reactions accompanying downhill lactose/H(+) symport catalyzed by the lactose permease of Escherichia coli (LacY) have been assessed using solid-supported membrane-based electrophysiology with improved time resolution. Rates of charge translocation generated by purified LacY reconstituted into proteoliposomes were analyzed over a pH range from 5.2 to 8.5, which allows characterization of two electrogenic steps in the transport mechanism: (i) a weak electrogenic reaction triggered by sugar binding and observed under conditions where H(+) translocation is abolished either by acidic pH or by a Glu325 --> Ala mutation in the H(+) binding site (this step with a rate constant of approximately 200 s(-1) for wild-type LacY leads to an intermediate proposed to represent an "occluded" state) and (ii) a major electrogenic reaction corresponding to 94% of the total charge translocated at pH 8, which is pH-dependent with a maximum rate of approximately 30 s(-1) and a pK of 7.5. This partial reaction is assigned to rate-limiting H(+) release on the cytoplasmic side of LacY during turnover. These findings together with previous electrophysiological results and biochemical-biophysical studies are included in an overall kinetic mechanism that allows delineation of the electrogenic steps in the reaction pathway.

The N- and C-terminal six-helix bundles of lactose permease (LacY) form a large internal cavity open on the cytoplasmic side and closed on the periplasmic side with a single sugar-binding site at the apex of the cavity near the middle of the molecule. During sugar/H+ symport, an outward-facing cavity is thought to open with closing of the inward-facing cavity so that the sugar-binding site is alternately accessible to either face of the membrane. In this communication, single-molecule fluorescence (Forster) resonance energy transfer is used to test this model with wild-type LacY and a conformationally restricted mutant. Pairs of Cys residues at the ends of two helices on the cytoplasmic or periplasmic sides of wild-type LacY and the mutant were labeled with appropriate donor and acceptor fluorophores, single-molecule fluorescence resonance energy transfer was determined in the absence and presence of sugar, and distance changes were calculated. With wild-type LacY, binding of a galactopyranoside, but not a glucopyranoside, results in a decrease in distance on the cytoplasmic side and an increase in distance on the periplasmic side. In contrast, with the mutant, a more pronounced decrease in distance and in distance distribution is observed on the cytoplasmic side, but there is no change on the periplasmic side. The results are consistent with the alternating access model and indicate that the defect in the mutant is due to impaired ligand-induced flexibility on the periplasmic side.

YidC of Echerichia coli, a member of the conserved Alb3/Oxa1/YidC family, is postulated to be important for biogenesis of membrane proteins. Here, we use as a model the lactose permease (LacY), a membrane transport protein with a known three-dimensional structure, to determine whether YidC plays a role in polytopic membrane protein insertion and/or folding. Experiments in vivo and with an in vitro transcription/translation/insertion system demonstrate that YidC is not necessary for insertion per se, but plays an important role in folding of LacY. By using the in vitro system and two monoclonal antibodies directed against conformational epitopes, LacY is shown to bind the antibodies poorly in YidC-depleted membranes. Moreover, LacY also folds improperly in proteoliposomes prepared without YidC. However, when the proteoliposomes are supplemented with purified YidC, LacY folds correctly. The results indicate that YidC plays a primary role in folding of LacY into its final tertiary conformation via an interaction that likely occurs transiently during insertion into the lipid phase of the membrane.