Structural Insights into the Mechanisms and Pharmacology of K2P Potassium Channels

Leak currents, defined as voltage and time independent flows of ions across cell membranes, are central to cellular electrical excitability control. The K_2P (KCNK) potassium channel class comprises an ion channel family that produces potassium leak currents that oppose excitation and stabilize the resting membrane potential in cells in the brain, cardiovascular system, immune system, and sensory organs. Due to their widespread tissue distribution, K_2Ps contribute to many physiological and pathophysiological processes including anesthesia, pain, arrythmias, ischemia, hypertension, migraine, intraocular pressure regulation, and lung injury responses. Structural studies of six homomeric K_2Ps have established the basic architecture of this channel family, revealed key moving parts involved in K_2P function, uncovered the importance of asymmetric pinching and dilation motions in the K_2P selectivity filter (SF) C-type gate, and defined two K_2P structural classes based on the absence or presence of an intracellular gate. Further, a series of structures characterizing K_2P:modulator interactions have revealed a striking polysite pharmacology housed within a relatively modestly sized (~70 kDa) channel. Binding sites for small molecules or lipids that control channel function are found at every layer of the channel structure, starting from its extracellular side through the portion that interacts with the membrane bilayer inner leaflet. This framework provides the basis for understanding how gating cues sensed by different channel parts control function and how small molecules and lipids modulate K_2P activity. Such knowledge should catalyze development of new K_2P modulators to probe function and treat a wide range of disorders.     

Positive indirect effects of top-predators on the behaviour and survival of juvenile fishes

Top-predators can suppress mesopredator behaviour through risk effects. However, there is limited understanding of whether such behavioural suppression can dampen the lethal and sub-lethal effects of mesopredators on bottom level prey. Here, we document a field experiment that examines whether the presence of top-predator cues (visual and chemical stimuli from a coral trout) can cascade to indirectly influence the behaviour and survival of juvenile fish prey of different species (Pomacentrus amboinensis and P. chrysurus) and size (small = 1.18 cm SL versus large = 1.32 cm SL). Results showed that habitat patches exposed to top-predator cues received fewer visits and foraging attacks from mesopredators, leading to higher space use (~ 46%), feeding rate (~ 95%) and survival (~ 67%) from juvenile fish prey. Survival was always higher for individuals of P. amboinensis and of large-size, independent of the presence or absence of top-predator cues. Our data indicate that predation risk from the top-predator indirectly favoured the persistence and behaviour of juvenile fishes by promoting risk-averse behavioural responses in mesopredators. This study underscores the behavioural mechanisms by which risk effects can cascade through the food web and highlights the consequences that harvesting top-predators may have on the replenishment of bottom prey populations.     

Best Management Practices for Trapping Furbearers in the United States

Humans have used wild furbearers for various purposes for thousands of years. Today, furbearers are sustainably used by the public for their pelts, leather, bones, glands, meat, or other purposes. In North America, contemporary harvest of furbearers has evolved along with trap technologies and societal concerns, and is now highly regulated and more closely coupled with harvest analysis and population monitoring. Traps and regulated trapping programs provide personal or cultural rewards that can also support conservation, and can assist with advancing ecological knowledge through research, protecting endangered species, restoring populations or habitats, protecting personal property, and enhancing public health and safety. However, animal welfare and trap selectivity remain important topics for furbearer management in North America, as they have for more than a century. A related international challenge to modern furbearer management came with the Wild Fur Regulation by the European Union, which passed in 1991. This regulation prohibited use of foothold traps in many European countries and the importation of furs and manufactured fur products to Europe from countries that allowed use of foothold traps or trapping methods that did not meet internationally agreed-upon humane trapping standards. To address existing national concerns and requirements of the Wild Fur Regulation, the United States and European Union signed a non-binding bilateral understanding that included a commitment by the United States to evaluate trap performance and advance the use of improved traps through development of best management practices (BMPs) for trapping. Our testing followed internationally accepted restraining-trap standards for quantifying injuries and capture efficiency, and we established BMP pass-fail thresholds for these metrics. We also quantified furbearer selectivity, and qualitatively assessed practicality and user safety for each trap, yielding overall species-specific performance profiles for individual trap models. We present performance data for 84 models of restraining traps (6 cage traps, 68 foothold traps, 9 foot-encapsulating traps, and 1 power-activated footsnare) on 19 furbearing species, or 231 trap-species combinations. We conducted post-mortem examinations on 8,566 furbearers captured by trappers. Of the 231 trap model-species combinations tested, we had sufficient data to evaluate 173 combinations, of which about 59% met all BMP criteria. Pooling species, cage traps produced the lowest average injury score (common injuries included tooth breakage), with minimal differences across other trap types; species-specific patterns were generally similar, with the exception of raccoons (Procyon lotor) for which foot-encapsulating traps performed better than other foot-restraining trap types. Padded-jaw foothold traps performed better than standard-jaw models for many species, though often similar to and occasionally worse than offset- or laminated-jaw models. Most traps we tested had high capture efficiency; only 5 (3%) failed BMP standards strictly because of poor efficiency. Average furbearer selectivity was high across all trap types we evaluated and was lowest for footsnares (88%) and highest for foot-encapsulating traps (99%). Mortality from trap-related injury in restraining traps we tested was very rare for furbearers (0.5% of animals). In over 230,000 trap-nights across a 21-year period, no individuals of a threatened or endangered species were captured. Of 9,589 total captures, 11% were non-furbearers, of which 83% were alive upon trap inspection; nearly all non-furbearer mortalities were birds, rabbits, or squirrels. Approximately 2% of total captures were feral or free-ranging dogs (Canis familiaris), of which none died or were deemed in need of veterinary care by either our technicians or the owners (if located). Similarly, 3% of total captures were feral or free-ranging cats (Felis catus); 2 were dead, and although locating potential owners was often impossible, none of the remaining cats were deemed in need of veterinary care by technicians or owners. Our results show that furbearer selectivity was high for all trap types evaluated, mortality or significant injury was very rare for domestic (or feral) animals, and the most potential for mortality or injury of non-furbearers was with smaller animals, a majority of which were squirrels and rabbits. Our results suggest that injury scores for a given trap-species combination are unlikely to vary significantly across states or regions of the United States, provided similar methods are employed. Our data also suggest that taxonomic affiliation and body-size groupings are correlated with injury scores, presumably through morphological, physiological, or behavioral adaptations or responses that influence injury potential during restraint; higher injury scores in foot-restraining trap types were more likely in smaller or more dexterous species, whereas injury scores were typically lowest for the felids we evaluated. For some species (e.g., American badger [Taxidea taxus], bobcat [Lynx rufus]), most restraining traps we tested met BMP standards, whereas few restraining traps we tested met standards for other species (e.g., muskrat [Ondatra zibethicus], striped skunk [Mephitis mephitis]). Comparison of our results with survey information collected during 2015 on trap use in the United States indicates that approximately 75% of all target furbearers harvested were taken in BMP-compliant traps, with another 10% taken in traps yet to be tested on that species. Future trap testing and development should focus on commonly used traps not yet tested on a species, species for which few passing traps currently pass BMP criteria, and trap models and modifications most likely to minimize trap injuries given a species morphology, physiology, and behavior. Outreach efforts should focus on general BMP awareness, discouraging use of traps that fail BMP standards for a given species, and public outreach on trapping. Restraining (and other) traps have evolved substantially in recent decades and offer numerous benefits to individuals, conservation, and society. However, continuing to address societal concerns remains a critical component of modern regulated trapping and furbearer management. Published trapping BMPs are regularly updated online and may include additional approved restraining and killing traps that were evaluated as part of testing by Canada. We will periodically update the trap performance tables and figures we presented and make them available online at the Association of Fish and Wildlife Agencies website. Published 2020. This article is a U.S. Government work and is in the public domain in the USA. Wildlife Monographs published by Wiley Periodicals LLC on behalf of The Wildlife Society.     

Summer diet of juvenile lake sturgeon reintroduced into the Genesee and St. Regis Rivers,New York USA

The restoration of threatened species involves understanding multiple aspects of the life history and ecology of the target species. One important consideration in the restoration of threatened species is feeding ecology. We examined the summer diet of reintroduced juvenile lake sturgeon (Acipenser fulvescens) in the Genesee (n = 119, ages 1 and 2) and the St. Regis rivers (n = 40, ages 1, 2, 5, 6, and 7) of New York State, USA, in the summer of 2005. The most common taxa consumed were Diptera (88% of prey biomass) in the Genesee River and Ephemeroptera (35% of prey biomass) and Diptera (29%) in the St. Regis River. Diptera: (Chironomidae) was the major prey taxon in both benthic communities. Selectivity analysis showed a positive selection of Diptera by Genesee River lake sturgeon and an avoidance of Diptera by St. Regis River lake sturgeon. The St. Regis River lake sturgeon showed avoidance of Coleoptera, but positive selection of Ephemeroptera, Trichoptera, and Oligochaeta. Results indicate that the reintroduced juvenile lake sturgeon are growing and successfully finding prey in these two rivers and appear to actively select specific prey types.     

人分泌型磷脂酶A2 GIIE的E54突变及其对底物选择性的影响

人分泌型磷脂酶A2 GIIE (Human secreted phospholipase A2 GIIE,hGIIE) 通过发挥酶催化作用,参与炎症反应和脂代谢过程。为了揭示hGIIE的底物选择机制,文中对hGIIE进行了定点突变,采用毕赤酵母Pichia pastoris重组表达突变体蛋白,然后通过阳离子交换和分子排阻两步法纯化蛋白,最后用等温微量热滴定仪测定酶活性。hGIIE的结构分析显示,氨基酸E54可能与GIIE的底物头基选择性有关,经过同源序列比对,拟将E54突变为丙氨酸 (A)、苯丙氨酸 (F) 和赖氨酸 (K)。突变体E54A、E54F和E54K在毕赤酵母组成型表达系统中实现重组表达,通过两步纯化,纯度达到90%以上。酶活性实验显示,突变体与底物1,2-二己酰卵磷脂 (1,2-dihexanoyl-sn- glycero-3-phosphocholine,DHPC) 和1,2-二己酰磷酸甘油 (1,2-dihexyl phosphate glycerol,DHPG) 的亲和力发生改变,其中突变体E54K与DHPG的Km值为突变前的0.39倍,亲和力明显增强;突变体E54F与DHPC的Km值为突变前的1.93倍,亲和力明显减弱。hGIIE的E54突变体蛋白与磷脂底物的亲和力发生明显改变,说明E54在hGIIE的磷脂底物选择性水解过程中起重要作用。     

Metal-based environment-sensitive MRI contrast agents

Interactions of paramagnetic metal complexes with their biological environment can modulate their magnetic resonance imaging (MRI) contrast–enhancing properties in different ways, and this has been widely exploited to create responsive probes that can provide biochemical information. We survey progress in two rapidly growing areas: the MRI detection of biologically important metal ions, such as calcium, zinc, and copper, and the use of transition metal complexes as smart MRI agents. In both fields, new imaging technologies, which take advantage of other nuclei (¹⁹F) and/or paramagnetic contact shift effects, emerge beyond classical, relaxation-based applications. Most importantly, in vivo imaging is gaining ground, and the promise of molecular MRI is becoming reality, at least for preclinical research.     

Exploring QSAR for CYP11B2 binding affinity and CYP11B2/CYP11B1 selectivity of diverse functional compounds using GFA and G/PLS techniques

A data set of a series of 132 structurally diverse compounds with cytochrome 11B2 and 11B1 (CYP11B2 and CYP11B1) enzyme inhibitory activities was subjected to molecular shape analysis to explore contributions of shape features as well as electronic, structural, and physicochemical parameters toward enzyme inhibitory activities, in search of appropriate molecular scaffolds with optimum substitutions for highly potent CYP11B2 inhibitors. Genetic function approximation (GFA) and genetic partial least squares (G/PLS) were used as chemometric tools for modeling, and the derived equations were of acceptable statistical quality considering both internal and external validation parameters (Q²: 0.514–0.659, R²_pred: 0.510–0.734). The G/PLS models with spline option for CYP11B2 and CYP11B1 inhibition and selectivity modeling appeared to be the best models based on r_m²_(overall) criterion. The study indicates the importance of the pyridinylnaphthalene and pyridylmethylene-indane scaffolds with less polar and electrophilic substituents for optimum CYP11B2 inhibitory activity and CYP11B2/CYP11B1 selectivity.     

Synthesis of pyrrolo[2,3-d]pyridazinones as potent,subtype selective PDE4 inhibitors

A series of pyrrolo[2,3-d]pyridazinones was synthesized and tested for their inhibitory activity on PDE4 subtypes A, B and D and selectivity toward Rolipram high affinity binding site (HARBS). New agents with interesting profile were reported; in particular compound 9e showed a good PDE4 subtype selectivity, being 8 times more potent (IC₅₀ = 0.32 μM) for PDE4B (anti-inflammatory) than for PDE4D (IC₅₀ = 2.5 μM), generally considered the subtype responsible for emesis. Moreover the ratio HARBS/PDE4B was particularly favourable for 9e (147), suggesting that the best arranged groups around the pyrrolopyridazinone core are an isopropyl at position-1, an ethoxycarbonyl at position-2, together with an ethyl group at position-6.

For compounds 8 and 15a the ability to inhibit TNFα production in PBMC was evaluated and the results are consistent with their PDE4 inhibitory activity.     

Structural elucidation and biological significance of 2-hydroxy-1-naphthaldehyde derived sulfonamides and their first row d-transition metal chelates

2-Hydroxy-1-naphthaldehyde derived sulfonamides and their first row d-transition metal chelates [cobalt (II), copper (II), nickel (II) and zinc (II)] have been synthesized and characterized. The nature of bonding and structure of all the compounds have been deduced from elemental analyses, infrared, ¹H NMR, ¹³C NMR, mass spectrometry, electronic spectra, magnetic susceptibility and conductivity measurements. An octahedral geometry has been suggested for all the complexes. The metal complexes were screened for their antibacterial and antifungal activities on different species of pathogenic bacteria and fungi and their biopotency has been discussed. The results of these studies revealed that all compounds showed moderate to significant antibacterial activity against all bacterial strains and good antifungal activity against various fungal strains. In-vitro cytotoxic properties of all the compounds against Artemia salina was also studies by brine shrimp bioassay.     

Design,synthesis and structure–activity relationship studies of novel and diverse cyclooxygenase-2 inhibitors as anti-inflammatory drugs

Abstract

Because of the pivotal role of cyclooxygenase (COX) in the inflammatory processes, non-steroidal anti-inflammatory drugs (NSAIDs) that suppress COX activities have been used clinically for the treatment of inflammatory diseases/syndromes; however, traditional NSAIDs exhibit serious side-effects such as gastrointestinal damage and hyper sensitivity owing to their COX-1 inhibition. Also, COX-2 inhibition-derived suppressive or preventive effects against initiation/proliferation/invasion/motility/recurrence/metastasis of various cancers/tumours such as colon, gastric, skin, lung, liver, pancreas, breast, prostate, cervical and ovarian cancers are significant. In this study, design, synthesis and structure–activity relationship (SAR) of various novel {2-[(2-, 3- and/or 4-substituted)-benzoyl, (bicyclic heterocycloalkanophenyl)carbonyl or cycloalkanecarbonyl]-(5- or 6-substituted)-1H-indol-3-yl}acetic acid analogues were investigated to seek and identify various chemotypes of potent and selective COX-2 inhibitors for the treatment of inflammatory diseases, resulting in the discovery of orally potent agents in the peripheral-inflammation model rats. The SARs and physicochemical properties for the analogues are described as significant findings. For graphical abstract: see Supplementary Material. (www.informahealthcare.com/enz)