Structure-Activity Relationships of Natural C-9-Methyl-Substituted 10-Membered Lactones and Their Semisynthetic Derivatives.

Fungal 10-membered lactones (TMLs), such as stagonolide A, herbarumin I, pinolidoxin, and putaminoxin, are promising candidates for the development of nature-derived herbicides. The aim of this study was to analyze the structure-activity relationships (SAR) of C-9-methyl-substituted TMLs with a multitarget bioassay approach to reveal compounds with useful (phytotoxic, entomotoxic, antimicrobial) or undesirable (cytotoxic) bioactivities. A new TML, stagonolide L (1), along with five known compounds (stagonolides D (2) and E (3), curvulides A (4) and B1/B2 (5a,b), and pyrenolide C (6)), were purified from cultures of the phytopathogenic fungus Stagonospora cirsii, and five semisynthetic derivatives of 3 and 4 (7-11) were obtained. The absolute configuration of 4 was revised to 2Z, 4S, 5S, 6R, and 9R. The identity of 5a,b and stagonolide H is discussed. The phytotoxicity of compound 4, the entomotoxicity of 5a,b, and nonselective toxicity of compound 6 are demonstrated. The latter confirms the hypothesis that the α,ß-unsaturated carbonyl group is associated with the high general toxicity of TML, regardless of its position in the ring and other substituents. The epoxide in compound 4 is important for phytotoxicity. The revealed SAR patterns will be useful for further rational design of TML-based herbicides including curvulide A analogs with a 4,5-epoxy group.

Cells with stochastically increased methyltransferase to restriction endonuclease ratio provide an entry for bacteriophage into protected cell population.

The action of Type II restriction-modification (RM) systems depends on restriction endonuclease (REase), which cleaves foreign DNA at specific sites, and methyltransferase (MTase), which protects host genome from restriction by methylating the same sites. We here show that protection from phage infection increases as the copy number of plasmids carrying the Type II RM Esp1396I system is increased. However, since increased plasmid copy number leads to both increased absolute intracellular RM enzyme levels and to a decreased MTase/REase ratio, it is impossible to determine which factor determines resistance/susceptibility to infection. By controlled expression of individual Esp1396I MTase or REase genes in cells carrying the Esp1396I system, we show that a shift in the MTase to REase ratio caused by overproduction of MTase or REase leads, respectively, to decreased or increased protection from infection. Consistently, due to stochastic variation of MTase and REase amount in individual cells, bacterial cells that are productively infected by bacteriophage have significantly higher MTase to REase ratios than cells that ward off the infection. Our results suggest that cells with transiently increased MTase to REase ratio at the time of infection serve as entry points for unmodified phage DNA into protected bacterial populations.

High-repetition-rate pulse generation via self-sustaining cross-gain modulation in a semiconductor-based laser.

We present a simple and efficient method for generating regular pulse trains with GHz pulse repetition rates in lasers based on semiconductor optical amplifiers (SOAs). This method enables pulse formation without active modulation or saturable absorption of the generated radiation. The method relies upon the self-sustaining cross-gain modulation which is achieved by adding the negative optical feedback (NOF) to a ring laser configuration. The resulting modulation of laser gain is shown to be restricted to the frequencies which match both the spacing of longitudinal laser modes and the highest peaks in the NOF-induced instability gain spectrum. This enables the reproducible stationary pulse generation at the strictly defined repetition rates. The feasibility of the method was confirmed by the stable generation of sub-nanosecond pulses at repetition rates up to 1.79 GHz in a SOA-based laser with a simple fiber cavity.

General characteristics of the influence of surfactants on the bacteriolytic activity of lysozyme based on the example of enzymatic lysis of Lactobacillus plantarum cells in the presence of Tween 21 and SDS.

The general characteristics of the effect of surfactants on the activity of lysozyme were demonstrated. The kinetics of bacterial cell lysis is consistent with the Michaelis-Menten equation and the presence of surfactants does not shift the pH-optimum of activity. Surfactants do not change the Km value but instead, affect the Vmax value. The experimental dependencies are well described by theoretical equations, which assume three surfactant binding sites on the lysozyme molecule. The dependencies of the activity of lysozyme on the surfactant concentration are either a step type (i.e., a higher plateau becomes a lower plateau), or a dependency with a maximum and continuation of the curve in the form of a plateau but with an increase in the surfactant concentration. It can be assumed that there is a mechanism for the regulation of lysozyme activity by an unknown natural factor that has a suitable hydrophobic radical capable of binding to the surface of lysozyme.

Investigation of the Dual-Stage Method of Active Er:YLF Laser Drug Delivery Through the Nail and Laser-Induced Transformations of the Drug Extinction Spectrum.

BACKGROUND AND OBJECTIVE: A novel dual-stage method for active laser drug delivery (DSLADD) in the treatment of nail diseases is being presented. This method includes sequentially performed microporation of the nail with submillisecond pulses of Er:YLF laser radiation through a layer of an aqueous solution of drug deposited on the nail surface (Stage 1) and exposure this layer to the same laser radiation to deliver drug under the nail plate (Stage 2). The delivery of methylene blue (MB) as one of the possible drugs in the treatment of nail diseases is investigated. The influence of the thickness of the MB layer, as well as the energy and number of applied laser pulses, on the rate of active laser delivery is discussed. To illustrate the possible effect of delivery on the drug delivered, special attention is paid to the deformation of the extinction spectrum of MB solution after laser irradiation. STUDY DESIGN/MATERIALS AND METHODS: Diode-pumped Er:YLF laser was used for DSLADD. The process of DSLADD under the nail plate was investigated using digital video microscopy. For different values of the thickness of MB solution layer applied to the nail plate and the energy of laser pulses, the number of laser pulses required to create a single through a microchannel in the nail plate and the number of laser pulses required to deliver the solution to the ventral side of the nail plate after its microporation were registered. The mass and the dose of MB solution penetrated under the nail plate, and the rate of MB solution delivery through a single microchannel was determined. Investigation of the influence of Er:YLF laser radiation parameters on the extinction spectrum of the drug was performed using a fiber spectrometer. The extinction spectra of the 0.001% aqueous solution of MB were recorded before and after exposure to a different number of Er:YLF laser pulses with the energy of 1-4 mJ. RESULTS: It was found that the minimum number of laser pulses required for active Er:YLF laser drug delivery under the nail corresponds to the MB layer thickness of 100 µm and the laser pulse energy of 4 mJ. It is shown that in this case, the rate of active laser delivery of MB solution reaches 0.26 ± 0.03 mg/pulse. The radiation of the Er:YLF laser affects the shape of the extinction spectrum of the aqueous solution of MВ, which is associated with the transition of the dye from the monomeric to dimeric state. Depending on the laser pulse energy, the fraction of a certain conformational state in the aqueous MB solution can decrease or increase, stimulating a possible change in its photodynamic and antiseptic activity. CONCLUSION: For the first time, a novel DSLADD through the nail has been described and investigated in vitro. It was demonstrated that at Er:YLF laser pulse repetition rate of f = 30 Hz, microporation of the nail plate and drug delivery through a single microchannel will be about 1.5 s. Lasers Surg. Med. © 2021 Wiley Periodicals LLC.

Model of Chronic Thromboembolic Pulmonary Hypertension in Rats Caused by Repeated Intravenous Administration of Partially Biodegradable Sodium Alginate Microspheres.

Chronic thromboembolic pulmonary hypertension (CTEPH) is a rare and life-threatening complication of pulmonary embolism. As existing animal models of CTEPH do not fully recapitulate complex disease pathophysiology, we report a new rat model for CTEPH evoked by repetitive embolization of the distal pulmonary artery branches with partially biodegradable alginate microspheres (MSs). MSs (180 ± 28 µm) were intravenously administered eight times at 4-day intervals; control animals received saline. The validity of the model was confirmed using transthoracic echocardiography, exercise testing, catheterization of the right ventricle, and histological examination of the lung and heart. The animals in the CTEPH group demonstrated a stable increase in right ventricular systolic pressure (RVSP) and decreased exercise tolerance. Histopathological examination revealed advanced medial hypertrophy in the small pulmonary arteries associated with fibrosis. The diameter of the main pulmonary artery was significantly larger in the CTEPH group than in the control group. Marinobufagenin and endothelin-1 serum levels were significantly elevated in rats with CTEPH. In conclusion, repetitive administration of alginate MSs in rats resulted in CTEPH development characterized by specific lung vasculature remodeling, reduced exercise tolerance, and a persistent rise in RVSP. The developed model can be used for pre-clinical testing of promising drug candidates.

Triggering of different pulsed regimes in fiber cavity laser by a waveguide electro-optic switch.

A novel practical method for electronic triggering of essentially different pulsed regimes in fiber cavity lasers is introduced. The method relies on electronic control of complementary transmission characteristics of a fiber-coupled LiNbO3 waveguide electro-optic switch (WEOS) which plays the role of the variable output coupler in a fiber cavity. The method was studied using a testbed laser configuration comprised of a semiconductor optical amplifier (SOA) and an all-fiber cavity. Modulation of the WEOS-based output coupling in the fast gain recovery configuration allowed not only high-quality mode locking and harmonic mode-locking at certain pulse repetition rates determined by the cavity round trip time, but it also allowed nanosecond pulsed output of the same quality to be yielded by cavity dumping at widely and continuously tunable repetition rate (ranging from kHz to MHz). Thus, WEOS-based electronically variable output coupling allows uniquely high flexibility for lasing regimes and characteristics within a single all-fiber cavity configuration.

Crystal Structure of the Labile Complex of IL-24 with the Extracellular Domains of IL-22R1 and IL-20R2.

Crystal structure of the ternary complex of human IL-24 with two receptors, IL-22R1 and IL-20R2, has been determined at 2.15 Å resolution. A crystallizable complex was created by a novel approach involving fusing the ligand with a flexible linker to the presumed low-affinity receptor, and coexpression of this construct in Drosophila S2 cells together with the presumed high-affinity receptor. This approach, which may be generally applicable to other multiprotein complexes with low-affinity components, was necessitated by the instability of IL-24 expressed by itself in either bacteria or insect cells. Although IL-24 expressed in Escherichia coli was unstable and precipitated almost immediately upon its refolding and purification, a small fraction of IL-24 remaining in the folded state was shown to be active in a cell-based assay. In the crystal structure presented here, we found that two cysteine residues in IL-24 do not form a predicted disulfide bond. Lack of structural restraint by disulfides, present in other related cytokines, is most likely reason for the low stability of IL-24. Although the contact area between IL-24 and IL-22R1 is larger than between the cytokine and IL-20R2, calculations show the latter interaction to be slightly more stable, suggesting that the shared receptor (IL-20R2) might be the higher-affinity receptor.

Enhanced calcium entry via activation of NOX/PKC underlies increased vasoconstriction induced by methylglyoxal.

Advanced glycation end-products (AGEs) play a pivotal role in macro- and micro-vascular diabetic complications. We investigated the mechanism by which methylglyoxal (an endogenous generator of AGEs) affects vascular contractility using the isolated artery technique. Contractile responses to vasoconstrictors phenylephrine (PE), angiotensin II (Ang II), vasopressin (VP) and KCl were measured in the isolated rat aorta following one-our exposure to methylglyoxal (50-200 µM). The perfused rat kidney was employed to confirm the effect of methylglyoxal on microvessels. Methylglyoxal-induced changes in cytosolic calcium were measured in the smooth muscle layer of the aorta with the calcium-sensing fluorophore Fluo-4 AM. Methylglyoxal significantly increased maximal contraction of the rat aorta to PE, Ang II and VP. Similar results were seen in response to the depolarizing vasoconstrictor KCl in macro and micro vessels. The methylglyoxal-induced increases in aortic contraction mediated by the agonist and KCl were endothelium independent. Methylglyoxal-induced increases in KCl-dependent aortic contraction were abolished after the removal of extracellular calcium or in the presence of the calcium channel blocker nifedipine. Incubation with the antioxidant N-acetyl-l-cysteine (NAC), apocynin (a nonselective NADPH oxidase (NOX) inhibitor) or chelerythrine (a protein kinase C (PKC) inhibitor) prior to methylglyoxal pre-treatment reversed the methylglyoxal-induced increases in the rat aortic contractility. In conclusion, the formation of AGEs increases vasoconstriction of both macro- and micro-vessels by increasing the voltage-activated calcium entry in vascular smooth muscles in a NOX and PKC dependent manner.

Correction: Distinct Roles of Type I and Type III Interferons in Intestinal Immunity to Homologous and Heterologous Rotavirus Infections.

[This corrects the article DOI: 10.1371/journal.ppat.1005600.].

Distinct Roles of Type I and Type III Interferons in Intestinal Immunity to Homologous and Heterologous Rotavirus Infections.

Type I (IFN-α/ß) and type III (IFN-λ) interferons (IFNs) exert shared antiviral activities through distinct receptors. However, their relative importance for antiviral protection of different organ systems against specific viruses remains to be fully explored. We used mouse strains deficient in type-specific IFN signaling, STAT1 and Rag2 to dissect distinct and overlapping contributions of type I and type III IFNs to protection against homologous murine (EW-RV strain) and heterologous (non-murine) simian (RRV strain) rotavirus infections in suckling mice. Experiments demonstrated that murine EW-RV is insensitive to the action of both types of IFNs, and that timely viral clearance depends upon adaptive immune responses. In contrast, both type I and type III IFNs can control replication of the heterologous simian RRV in the gastrointestinal (GI) tract, and they cooperate to limit extra-intestinal simian RRV replication. Surprisingly, intestinal epithelial cells were sensitive to both IFN types in neonatal mice, although their responsiveness to type I, but not type III IFNs, diminished in adult mice, revealing an unexpected age-dependent change in specific contribution of type I versus type III IFNs to antiviral defenses in the GI tract. Transcriptional analysis revealed that intestinal antiviral responses to RV are triggered through either type of IFN receptor, and are greatly diminished when receptors for both IFN types are lacking. These results also demonstrate a murine host-specific resistance to IFN-mediated antiviral effects by murine EW-RV, but the retention of host efficacy through the cooperative action by type I and type III IFNs in restricting heterologous simian RRV growth and systemic replication in suckling mice. Collectively, our findings revealed a well-orchestrated spatial and temporal tuning of innate antiviral responses in the intestinal tract where two types of IFNs through distinct patterns of their expression and distinct but overlapping sets of target cells coordinately regulate antiviral defenses against heterologous or homologous rotaviruses with substantially different effectiveness.

Mu-driven transposition of recombinant mini-Mu unit DNA in the Corynebacterium glutamicum chromosome.

A dual-component Mu-transposition system was modified for the integration/amplification of genes in Corynebacterium. The system consists of two types of plasmids: (i) a non-replicative integrative plasmid that contains the transposing mini-Mu(LR) unit bracketed by the L/R Mu ends or the mini-Mu(LER) unit, which additionally contains the enhancer element, E, and (ii) an integration helper plasmid that expresses the transposition factor genes for MuA and MuB. Efficient transposition in the C. glutamicum chromosome (≈ 2 × 10-4 per cell) occurred mainly through the replicative pathway via cointegrate formation followed by possible resolution. Optimizing the E location in the mini-Mu unit significantly increased the efficiency of Mu-driven intramolecular transposition-amplification in C. glutamicum as well as in gram-negative bacteria. The new C. glutamicum genome modification strategy that was developed allows the consequent independent integration/amplification/fixation of target genes at high copy numbers. After integration/amplification of the first mini-Mu(LER) unit in the C. glutamicum chromosome, the E-element, which is bracketed by lox-like sites, is excised by Cre-mediated fashion, thereby fixing the truncated mini-Mu(LR) unit in its position for the subsequent integration/amplification of new mini-Mu(LER) units. This strategy was demonstrated using the genes for the citrine and green fluorescent proteins, yECitrine and yEGFP, respectively.

Layout of NALM fiber laser with adjustable peak power of generated pulses.

The Letter proposes a new layout of a passively mode-locked fiber laser based on a nonlinear amplifying loop mirror (NALM) with two stretches of active fiber and two independently controlled pump modules. In contrast with conventional NALM configurations using a single piece of active fiber that yields virtually constant peak power, the proposed novel laser features larger than a factor of 2 adjustment range of peak power of generated pulses. The proposed layout also provides independent adjustment of duration and peak power of generated pulses as well as power-independent control of generated pulse spectral width impossible in NALM lasers with a single piece of active fiber.

Mode-locked long fibre master oscillator with intra-cavity power management and pulse energy > 12 µJ.

Combined lengthening of the cavity of a passive mode-locked fibre master oscillator and implementation of a new concept of intra-cavity power management led to achievement of a record-high pulse energy directly at the output of the mode-locked fibre master oscillator (without any subsequent amplification) exceeding 12 µJ. Output powers at the level of > 12 µJ obtainable from a long-cavity mode-locked fibre master oscillator open new possibilities of application of all pulse types that can be generated in such oscillators.

Solid-State and Solution Metallophilic Aggregation of a Cationic [Pt(NCN)L](+) Cyclometalated Complex.

The noncovalent intermolecular interactions (π-π stacking, metallophilic bonding) of the cyclometalated complexes [Pt(NCN)L](+)X(-) (NCN = dipyridylbenzene, L = pyridine (1), acetonitrile (2)) are determined by the steric properties of the ancillary ligands L in the solid state and in solution, while the nature of the counterion X(-) (X(-) = PF6(-), ClO4(-), CF3SO3(-)) affects the molecular arrangement of 2·X in the crystal medium. According to the variable-temperature X-ray diffraction measurements, the extensive Pt···Pt interactions and π-stacking in 2·X are significantly temperature-dependent. The variable concentration (1)H and diffusion coefficients NMR measurements reveal that 2·X exists in the monomeric form in dilute solutions at 298 K, while upon increase in concentration [Pt(NCN)(NCMe)](+) cations undergo the formation of the ground-state oligomeric aggregates with an average aggregation number of ∼3. The photoluminescent characteristics of 1 and 2·X are largely determined by the intermolecular aggregation. For the discrete molecules the emission properties are assigned to metal perturbed IL charge transfer mixed with some MLCT contribution. In the case of oligomers 2·X the luminescence is significantly red-shifted with respect to 1 and originates mainly from the (3)MMLCT excited states. The emission energies depend on the structural arrangement in the crystal and on the complex concentration in solution, variation of which allows for the modulation of the emission color from greenish to deep red. In the solid state the lability of the ligands L leads to vapor-induced reversible transformation 1 ↔ 2 that is accompanied by the molecular reorganization and, consequently, dramatic change of the photophysical properties. Time-dependent density functional theory calculations adequately support the models proposed for the rationalization of the experimental observations.

Receptor tyrosine kinases, TYRO3, AXL, and MER, demonstrate distinct patterns and complex regulation of ligand-induced activation.

TYRO3, AXL, and MER receptors (TAMs) are three homologous type I receptor-tyrosine kinases that are activated by endogenous ligands, protein S (PROS1) and growth arrest-specific gene 6 (GAS6). These ligands can either activate TAMs as soluble factors, or, in turn, opsonize phosphatidylserine (PS) on apoptotic cells (ACs) and serve as bridging molecules between ACs and TAMs. Abnormal expression and activation of TAMs have been implicated in promoting proliferation and survival of cancer cells, as well as in suppressing anti-tumor immunity. Despite the fact that TAM receptors share significant similarity, little is known about the specificity of interaction between TAM receptors and their ligands, particularly in the context of ACs, and about the functional diversity of TAM receptors. To study ligand-mediated activation of TAMs, we generated a series of reporter cell lines expressing chimeric TAM receptors. Using this system, we found that each TAM receptor has a unique pattern of interaction with and activation by GAS6 and PROS1, which is also differentially affected by the presence of ACs, PS-containing lipid vesicles and enveloped virus. We also demonstrated that γ-carboxylation of ligands is essential for the full activation of TAMs and that soluble immunoglobulin-like TAM domains act as specific ligand antagonists. These studies demonstrate that, despite their similarity, TYRO3, AXL, and MER are likely to perform distinct functions in both immunoregulation and the recognition and removal of ACs.

Radiation build-up in laminar and turbulent regimes in quasi-CW Raman fiber laser.

We study the radiation build-up in laminar and turbulent generation regimes in quasi-CW Raman fiber laser. We found the resulted spectral shape and generation type is defined by the total spectral broadening/narrowing balance over laser cavity round-trip, which is substantially different in different regimes starting from first round-trips of the radiation build-up. In turbulent regime, the steady-state is reached only after a few round-trips, while in the laminar regime the laser approaches the equilibrium spectrum shape asymptotically.

Cascaded SRS of single- and double-scale fiber laser pulses in long extra-cavity fiber.

This work presents, for the first time, the results of studies of stimulated Raman scattering (SRS) in 1.2-km P2O5-doped silica fiber of radiation of single- and double-scale picosecond pulses generated in a fiber master oscillator and amplified in a one-stage fiber amplifier. Shown are differences in supercontinuum spectra composed of several Stokes components when pumped with pulses of different structure. More efficient Raman transformation of double-scale pulses was identified, leading to broader supercontinuum spectra.

Efficiency of non-linear frequency conversion of double-scale pico-femtosecond pulses of passively mode-locked fiber laser.

For the first time we report the results of both numerical simulation and experimental observation of second-harmonic generation as an example of non-linear frequency conversion of pulses generated by passively mode-locked fiber master oscillator in different regimes including conventional (stable) and double-scale (partially coherent and noise-like) ones. We show that non-linear frequency conversion efficiency of double-scale pulses is slightly higher than that of conventional picosecond laser pulses with the same energy and duration despite strong phase fluctuations of double-scale pulses.

Peculiarities of charge compensation in lithium-doped hydroxyapatite.

Hydroxyapatite (HA) remains one of the most popular materials for various biomedical applications and its fields of application have been expanding. Lithium (Li+) is a promising candidate for modifying the biological behavior of HA. Li+ is present in trace amounts in the human body as an alkaline and bioelectric material. At the same time, the introduction of Li+ into the HA structure required charge balance compensation due to the difference in oxidation degree, and the scheme of this compensation is still an open question. In the present work, the results of the theoretical and experimental study of the Li+-doped HA synthesis are presented. According to X-ray diffraction data, Fourier transform infrared spectroscopy as well as the combination of electron paramagnetic resonance methods, the introduction of Li+ in the amount up to 0.05 mol% resulted in the preservation of the HA structure. Density functional theory calculations show that Li+ preferentially incorporates into the Ca (1) position with a small geometry perturbation. The less probable positioning in the Ca (2) position leads to a drastic perturbation of the anion channel.