Selective covalent targeting of GPX4 using masked nitrile-oxide electrophiles.

We recently described glutathione peroxidase 4 (GPX4) as a promising target for killing therapy-resistant cancer cells via ferroptosis. The onset of therapy resistance by multiple types of treatment results in a stable cell state marked by high levels of polyunsaturated lipids and an acquired dependency on GPX4. Unfortunately, all existing inhibitors of GPX4 act covalently via a reactive alkyl chloride moiety that confers poor selectivity and pharmacokinetic properties. Here, we report our discovery that masked nitrile-oxide electrophiles, which have not been explored previously as covalent cellular probes, undergo remarkable chemical transformations in cells and provide an effective strategy for selective targeting of GPX4. The new GPX4-inhibiting compounds we describe exhibit unexpected proteome-wide selectivity and, in some instances, vastly improved physiochemical and pharmacokinetic properties compared to existing chloroacetamide-based GPX4 inhibitors. These features make them superior tool compounds for biological interrogation of ferroptosis and constitute starting points for development of improved inhibitors of GPX4.

Habitat and microclimate affect the resting site selection of mosquitoes.

Mosquitoes (Diptera: Culicidae) use certain resting sites during their inactive phase. The microclimatic conditions of these resting sites might affect their physiology and vectorial capacity. In this study, we combined a field and a laboratory study to investigate the natural resting site and temperature preferences of mosquitoes. The field study was conducted at a forest close to Oldenburg (Lower Saxony, Germany) from May to October 2018. Mosquitoes were collected in five different natural habitats with seven replicates each. Temperature was recorded hourly at each site. Significantly more mosquitoes were collected in deadwood (predominantly Culiseta morsitans/fumipennis) and shaded herb layer (predominantly Aedes species) compared to unshaded herb layer or broadleaf and coniferous trees. GLMMs revealed resting site habitats as the best predictor to explain the observed preference patterns, but microclimatic conditions are also involved in mosquito resting site selection. Most mosquitoes were collected at resting sites with relatively colder and more stable temperatures. In concert, laboratory choice experiments with a thermal gradient ring demonstrated that Cs. morsitans/fumipennis avoid temperatures over 30 °C. Understanding the small-scaled resting site preferences and the related microclimatic conditions can improve mosquito collection techniques and refine the prediction of mosquito-borne pathogen transmission.

Linking tree genetics and stream consumers: isotopic tracers elucidate controls on carbon and nitrogen assimilation.

Leaf litter provides an important nutrient subsidy to headwater streams, but little is known about how tree genetics influence energy pathways from litter to higher trophic levels. Despite the charge to quantify carbon (C) and nitrogen (N) pathways from decomposing litter, the relationship between litter decomposition and aquatic consumers remains unresolved. We measured litter preference (attachments to litter), C and N assimilation rates, and growth rates of a shredding caddisfly (Hesperophylax magnus, Limnephilidae) in response to leaf litter of different chemical and physical phenotypes using Populus cross types (P. fremontii, P. angustifolia, and F1 hybrids) and genotypes within P. angustifolia. We combined laboratory mesocosm studies using litter from a common garden with a field study using doubly labeled litter (13 C and 15 N) grown in a greenhouse and incubated in Oak Creek, Arizona, USA. We found that, in the lab, shredders initially chose relatively labile (low lignin and condensed tannin concentrations, rapidly decomposing) cross type litter, but preference changed within 4 d to relatively recalcitrant (high lignin and condensed tannin concentrations, slowly decomposing) litter types. Additionally, in the lab, shredder growth rates were higher on relatively recalcitrant compared to labile cross type litter. Over the course of a three-week field experiment, shredders also assimilated more C and N from relatively recalcitrant compared to labile cross type litter. Finally, among P. angustifolia genotypes, N assimilation by shredders was positively related to litter lignin and C:N, but negatively related to condensed tannins and decomposition rate. C assimilation was likewise positively related to litter C:N, and also to litter %N. C assimilation was not associated with condensed tannins or lignin. Collectively, these findings suggest that relatively recalcitrant litter of Populus cross types provides more nutritional benefit, in terms of N fluxes and growth, than labile litter, but among P. angustifolia genotypes the specific trait of litter recalcitrance (lignin or tannins) determines effects on C or N assimilation. As shredders provide nutrients and energy to higher trophic levels, the influence of these genetically based plant decomposition pathways on shredder preference and performance may affect community and food web structure.

Emergence of Culicoides obsoletus group species from farm-associated habitats in Germany.

Biting midges of the genus Culicoides (Diptera: Ceratopogonidae) may transmit several arboviruses to ruminant livestock. The species of the Obsoletus group are considered to be among the most important vectors of bluetongue virus (BTV) in northern Europe. As agricultural environments offer suitable habitats for the development of their immature stages, the emergence of adult Culicoides from potential breeding sites was investigated at 20 cattle farms throughout Germany in 2012 and 2013. In analyses of species-specific habitat preferences and relationships between Culicoides abundance in breeding substrates and their physicochemical characteristics, dungheaps emerged as the most important substrate for the development of Culicoides obsoletus sensu stricto (s.s.) (Meigen), whereas Culicoides chiopterus (Meigen) and Culicoides dewulfi Goetghebuer were generally restricted to cowpats. A decreasing pH value was associated with a higher abundance or a higher probability of observing these three species. Furthermore, the abundance of C. obsoletus s.s. was positively related to increasing moisture. Dungheaps were very productive breeding sites for this species and are therefore suggested as a target for potential control measures.

Distribution of individual members of the mosquito Anopheles maculipennis complex in Germany identified by newly developed real-time PCR assays.

Owing to their role as vectors of malaria parasites, species of the Anopheles maculipennis complex (Diptera: Culicidae) Meigen were intensively studied in the past, but with the disappearance of malaria in Germany in the middle of the last century, the interest in this field of research declined. A comprehensive ecological analysis of the current species distribution for Germany is lacking. Between 2010 and 2013, a total of 1445 mosquitoes of the An. maculipennis complex were collected at 72 different sites in Germany. The samples comprise 722 single individuals as well as 723 individuals in 90 pools of up to 25 mosquitoes. All samples were analysed with newly developed species-specific qPCR assays for the identification of the four German species using nucleotide differences within the internal transcribed spacer 2 (ITS2) ribosomal DNA. All gathered data were used for species distribution modelling. The overall prevalence of An. messeae s.l. was highest with 98.89% of all pools; An. daciae with 6.93% of all individuals and An. messeae s.s. with 69.53%. The prevalence of the other two species was relatively low: An. maculipennis s.s. with 13.30% of all individuals (6.67% of all pools) and An. atroparvus with 1.80% of all individuals (1.11% of all pools).

Impact of experimental flooding on larvae and pupae of dung-breeding Culicoides.

Culicoides biting midges (Diptera: Ceratopogonidae) spend the greatest part of their life in the larval stage. However, knowledge on the immature stages and the impact of abiotic factors on their development is still poor. Therefore, we investigated the effect of flooding on the larvae and pupae of Culicoides chiopterus (Meigen, 1830) and C. dewulfi Goetghebuer, 1936. In water, the larvae of both species showed head-to-tail flexions and sinuous flexions, at slow rates, but were not able to swim. Flooding of larvae for 24 h did not affect the number of emerging adults; flooding of pupae significantly reduced the emergence rate of C. chiopterus, compared to the control group, while C. dewulfi was not affected. Pupae were not able to float and no pupae survived flooding for 10 days. After flooding of larvae for 10 days, 50 % of C. chiopterus and 4 % of C. dewulfi completed the pre-adult development. During this treatment, 84 % of C. chiopterus and 48 % of C. dewulfi larvae pupated in water.

Divergent Total Syntheses of Rhodomyrtosones A and B.

Herein, we report total syntheses of the tetramethyldihydroxanthene natural product rhodomyrtosone B and the related bis-furan ß-triketone natural product rhodomyrtosone A. Nickel-(II)-catalyzed 1,4-conjugate addition of an α-alkylidene-ß-dicarbonyl substrate was developed to access the congener rhodomyrtosone B, and oxygenation of the same monoalkylidene derivative followed by cyclization was employed to obtain the bis-furan natural product rhodomyrtosone A.

Enantioselective total synthesis of (-)-maoecrystal V.

The enantioselective synthesis of maoecrystal V, a cytotoxic polycyclic diterpene, is described. Key reactions in the synthesis include an intramolecular Heck reaction, an oxidative cycloetherification, and an intermolecular Diels-Alder reaction to forge the carbocyclic core in a concise and stereoselective manner. Late-stage amine and C-H oxidation is used to install the final functional groups required to complete the synthesis.

The total synthesis of Isodon diterpenes.

Families of structurally related molecules often provide stimulating targets for organic chemists that are engaged in the development of new methods and strategies for natural product synthesis. While typically focused on specific molecules, these synthetic investigations often lead to generalizable concepts and significant opportunities for learning in a greater sense. Historically well-investigated families of natural products, such as the prostanoids, indole alkaloids, and macrolide antibiotics, provide ample evidence for the enduring value of these collective activities. In this Minireview, we turn our attention to the polycyclic family of diterpenes isolated from the Isodon genus of plants and provide an account of the recent methods and strategies utilized for their total synthesis.

Evaluation of 'east-to-west' ether-forming strategies for the total synthesis of maoecrystal V.

Model systems that evaluated different approaches to construct the central ether ring of maoecrystal V are described. Our first model systems attempted the ether formation using C-H functionalization reactions, which led to interesting rearrangements but none of the desired ether product. An intramolecular conjugate addition strategy was then explored that successfully formed the targeted C-O bond but resulted in the undesired stereochemistry.

Effects of Hibernation Site, Temperature, and Humidity on the Abundance and Survival of Overwintering Culex pipiens pipiens and Anopheles messeae (Diptera: Culicidae).

Knowledge of the hibernation site preferences and the factors which influence winter survival in these hibernation sites may enhance understanding of mosquito population dynamics after winter and how arboviruses persist in temperate regions. Our study quantified the number of adult overwintering mosquitoes in cellars and aboveground constructions and analyzed survival rates in relation to the environmental conditions in these sites. During the winters 2016/2017 and 2018/2019, 149 different constructions in Northwest Germany were sampled for mosquitoes. Mosquitoes were detected in 44% of the cellars and in 33% of the aboveground constructions. Culex p. pipiens Linnaeus was the most abundant species in cellars, whereas high numbers of Anopheles messeae Falleroni were collected from a single barn. Subsequently, an enclosure study was conducted during 2019/2020. Overwintering field-collected Cx. p. pipiens and An. messeae were divided into groups with or without fructose availability, and placed in cages with different man-made hibernations sites, where temperature and relative humidity were recorded hourly. For both species, increasing mean temperatures (5-16°C) but not mean relative humidity (58-94%) were correlated with winter mortality rates of the mosquitoes. The lipid measurements were greater and mortality rates were lower when both species were provided fructose. Larger specimens (determined by wing length) stored more lipids, and in Cx. p pipiens, but not in An. messeae, survival probability of large specimens was significantly greater than for small females. Mosquitoes showed a distinct pattern in the selection of overwintering sites, while temperature was an important driver for survival.

Enantioselective total synthesis and confirmation of the absolute and relative stereochemistry of streptorubin B.

The enantioselective total synthesis of the pyrrolophane natural product streptorubin B is described. Key steps in the concise route include the application of a one-pot enantioselective aldol cyclization/Wittig reaction and an anionic oxy-Cope rearrangement to forge the crucial 10-membered ring. Comparisons between CD spectra of synthetic and natural samples of streptorubin B coupled with X-ray crystallography allowed for the determination of the absolute stereochemistry of this natural product for the first time. These studies also provided unambiguous proof of the relative configuration between the butyl side chain and the bispyrrole subunit. Additional studies revealed a novel atropstereoselective Paal-Knorr pyrrole condensation and provided fundamental experimental insight into the barrier for atropisomerization of the natural product.

Geometric morphometric wing analysis represents a robust tool to identify female mosquitoes (Diptera: Culicidae) in Germany.

Accurate species identification is the prerequisite to assess the relevance of mosquito specimens, but is often hindered by missing or damaged morphological features. The present study analyses the applicability of wing geometric morphometrics as a low-cost and practical alternative to identify native mosquitoes in Germany. Wing pictures were collected for 502 female mosquitoes of five genera and 19 species from 80 sampling sites. The reliable species identification based on interspecific wing geometry of 18 landmarks per specimen was tested. Leave-one-out cross validation revealed an overall accuracy of 99% for the genus and 90% for the species identification. Misidentifications were mainly due to three pairings of Aedes species: Aedes annulipes vs. Aedes cantans, Aedes cinereus vs. Aedes rossicus and Aedes communis vs. Aedes punctor. Cytochrome oxidase subunit I (COI) gene region was sequenced to validate the morphological and morphometric identification. Similar to the results of the morphometric analysis, the same problematic three Aedes-pairs clustered, but most other species could be well separated. Overall, our study underpins that morphometric wing analysis is a robust tool for reliable mosquito identification, which reach the accuracy of COI barcoding.

Comparison of emergence traps of different shape and translucency in the trapping of Culicoides (Diptera: Ceratopogonidae).

Various types of emergence traps are available for investigations of the breeding habitats of Culicoides (Diptera: Ceratopogonidae). In order to assess the potential impact of the trap design on the trapping success, we compared the efficiency of opaque and white (more translucent) emergence traps and two trap shapes (cone-shaped and quadratic), to sample Culicoides emerging from cowpats. Significantly higher numbers of Culicoides chiopterus and Culicoides dewulfi were trapped with opaque traps, while there was no obvious effect of the trap shape. There were no distinct differences in the microclimate among different trap types.

Impact of freezing on the emergence of Culicoides chiopterus and Culicoides dewulfi from bovine dung.

The emergence of Culicoides chiopterus (Meigen), 1830 and C. dewulfi Goetghebuer, 1936 (Diptera: Ceratopogonidae) from cowpats in northwestern Germany was investigated. In order to investigate the survival of both species at low temperatures, cowpat subsamples were frozen for 48h at -18 and -21°C. Emergence from frozen and non-frozen samples was compared. The number of emerging adults of C. chiopterus from samples frozen at -18°C was greatly reduced and no emergence was observed from samples frozen at -21°C. No adult C. dewulfi emerged from frozen samples, suggesting this species is less resistant to these temperatures, compared to C. chiopterus.

Cardiac decompensation following verapamil therapy in infants with supraventricular tachycardia.

Three infants with supraventricular tachycardia and congestive cardiac failure were given verapamil intravenously. In two of the infants, the rhythm was converted to sinus, but the third infant required direct current cardioversion. However, each infant demonstrated hemodynamic decompensation shortly after verapamil administration and required cardiopulmonary resuscitation. All three patients were stabilized and their tachycardia was controlled with digoxin. All three were doing well at the time of follow-up evaluation, and there was no evidence of structural heart disease. The risk of cardiac decompensation in infants with supraventricular tachycardia and congestive cardiac failure should be kept in mind prior to administration of verapamil. Alternative methods for conversion of supraventricular tachycardia should be considered.

Cardiovascular manifestations in the Larsen syndrome.

The Larsen syndrome consists of a skeletal dysplasia with multiple joint dislocations and a characteristic facies. The basis of the abnormalities is felt to be a generalized mesenchymal disorder involving connective tissues. More than 80 cases have been reported in the literature with isolated reports of congenital cardiac septal defects and acquired abnormalities of the aorta and mitral valve. A case with marked aortic dilation and insufficiency as well as an aneurysm of the ductus arteriosus is presented. The aortic lesions are similar to those described in other connective tissue disorders, particularly the Marfan syndrome. Previous reports of ductal aneurysms have not revealed an association with connective tissue disorders, but have described a significant morbidity. In summary, patients with the Larsen syndrome are likely to have cardiac lesions similar to those classically associated with the Marfan syndrome; these patients deserve a careful investigation for cardiac anomalies. These aortic lesions may be as prognostically significant as cardiac lesions in the Marfan syndrome.

Double-outlet right ventricle and severe systemic outflow tract hypoplasia.

Double-outlet right ventricle and severe systemic outflow tract hypoplasia comprises a subset of patients in whom total correction or palliation requires complex surgical procedures in the neonatal period. Our experience with 3 patients illustrates the difficulties associated with treatment and suggests possible surgical options for this otherwise lethal variant of the Taussig-Bing syndrome.

Transcatheter closure of patent ductus arteriosus with the Amplatzer duct occluder: short-term follow-up.

BACKGROUND: Transcatheter closure of the patent ductus arteriosus is a well established procedure. The objective of this study was to assess the effectiveness and the safety of the Amplatzer duct occluder. METHODS AND RESULTS: Occlusion of the patent ductus arteriosus was attempted in 23 patients. The median weight was 11.7 Kg (range, 5 kg - 42.4 kg) with a mean ductus diameter of 3.7 mm (range, 1.6 - 7.2 mm). The immediate closure rate was 86% with a closure rate of 100% at 6 months, 1 year and 2 years following device placement. There was one device embolization that occurred immediately following device placement. No patient had aortic narrowing or pulmonary artery stenosis following the procedure. CONCLUSIONS: The Amplatzer duct occluder is safe and effective in the closure of a patent ductus arteriosus up to 7.2 mm in diameter. Selecting a device at least 12 mm larger than the minimal ductal diameter can minimize embolization.

Assessment of the abundance of Culicoides chiopterus and Culicoides dewulfi in bovine dung: a comparison of larvae extraction techniques and emergence traps.

We compared two larvae extraction methods involving sugar-flotation and an adapted Berlese funnel-extraction with emergence traps. This was done in order to analyse the colonisation of cowpats by Culicoides chiopterus (Meigen 1830) and Culicoides dewulfi (Goetghebuer 1936) (Diptera: Ceratopogonidae) and to gain information on the comparability and efficacy of these three methods. With all three methods, a considerable number of individuals was obtained. Significantly more individuals were obtained via sugar-flotation and Berlese funnel-extraction compared to the emergence traps. These differences, likely due to natural mortality and sample processing are discussed. We recommend Berlese as an efficacious method for extracting Culicoides larvae from bovine dung. It produces data rapidly and extracted larvae are viable. In comparison with Berlese, slightly more larvae were obtained by sugar-flotation, but this method was very labour intensive.