Identification of the native Torpedo californica nicotinic acetylcholine receptor's glycan composition after a multi-step sequential purification method using MALDI-ToF MS.

The Cys-loop pentameric ligand-gated ion channels comprise a dynamic group of proteins that have been extensively studied for decades, yielding a wealth of findings at both the structural and functional levels. The nicotinic acetylcholine receptor (nAChR) is no exception, as it is part of this large protein family involved in proper organismal function. Our efforts have successfully produced a highly pure nAChR in detergent complex (nAChR-DC), enabling more robust studies to be conducted on it, including beginning to experiment with high-throughput crystallization. Our homogeneous product has been identified and extensively characterized with 100% identity using Nano Lc MS/MS and MALDI ToF/ToF for each nAChR subunit. Additionally, the N-linked glycans in the Torpedo californica-nAChR (Tc-nAChR) subunits have been identified. To study this, the Tc-nAChR subunits were digested with PNGase F and the released glycans were analyzed by MALDI-ToF. The MS results showed the presence of high-mannose N-glycan in all native Tc-nAChR subunits. Specifically, the oligommanose population Man8-9GlcNac2 with peaks at m/z 1742 and 1904 ([M + Na]+ ions) were observed.

Evaluating the Suitability of Linear and Nonlinear Regression Approaches for the Langmuir Adsorption Model as Applied toward Biomass-Based Adsorbents: Testing Residuals and Assessing Model Validity.

Regression analysis is a powerful tool in adsorption studies. Researchers often favor linear regression for its simplicity when fitting isotherm models, such as the Langmuir equation. Validating regression assumptions is crucial to ensure that the model accurately represents the data and allows appropriate inferences. This study provides a detailed examination of assumption checking in the context of adsorption studies while simultaneously evaluating the robustness of linear regression methods for fitting the Langmuir equation to isotherm data from 2,4-dichlorophenol (DCP) adsorption onto various biomass-based adsorbents and activated carbon. Different linearized Langmuir equations (Hanes-Woolf, Lineweaver-Burk, Eadie-Hofstee, and Scatchard) were compared to nonlinear regression, and each method was validated by rigorous residual checking. This included visual plots of residuals as well as statistical tests, including the Durbin-Watson test for autocorrelation (independence), the Shapiro-Wilk test for normality, and the White test for homoscedasticity. Key findings indicate that the Hanes-Woolf (type 1) and Lineweaver-Burk (type 2) linearizations were the best for most biomass adsorbents studied and that Eadie-Hofstee (type 3) and Scatchard (type 4) were generally invalid due to the negative parameters or assumption violations. For activated carbon, all linearization methods were unsuitable due to independence violations. In the case of nonlinear regression, there were no major assumption violations for all of the adsorbents. Symbolic regression identified the Langmuir equation only for activated carbon (AC). This study revealed shortcomings in relying solely on linearized Langmuir models. A proposed workflow recommends using nonlinear or weighted nonlinear regression, starting with Hanes-Woolf or Lineweaver-Burk linearization results as initial values for parameter estimation. If assumptions remain violated with nonlinear techniques, novel methods such as symbolic regression should be employed. This advanced regression technique can improve adsorption models' accuracy and predictive behavior without the stringent need for assumption checking. Symbolic regression can also aid in understanding mechanisms of novel adsorbents.

May Measurement Month 2021: an analysis of blood pressure screening results from Venezuela.

Cardiovascular diseases continue to be the main cause of death in Venezuela, and hypertension is the principal risk factor. The May Measurement Month (MMM) campaign is a global initiative aimed to raising awareness of hypertension, which has been conducted in Venezuela since 2017. May Measurement Month 2021 included 46 732 participants with a mean age of 56.4 years (SD 14.4), 57.9% of whom were female. The percentage with hypertension was 60.3% (57.9% of females and 63.7% of males), 82.3% (84.8% of females and 79.2% of males) were aware, and 80.2% were taking antihypertensive medication. Of those on antihypertensive medication, 44.2% (41.2% of females and 48.2% of males) were not controlled [blood pressure (BP) ≥ 140/≥90 mmHg], with 61.3% receiving one drug, 30.0% two drugs, and 8.7% three or more drugs. 87.5% of those on treatment reported taking it regularly. Conditions associated with higher BP levels include fewer years of education, having a previous diagnosis of hypertension, and women who were hypertensive during a previous pregnancy. Physical activity and pregnancy were conditions associated with lower BP levels. A previous positive COVID-19 test was reported in 11.1%, and one or more COVID-19 vaccinations reported in 22.7% of participants. Of those on antihypertensive medication, 78.8% reported their treatment was not affected by COVID-19. Results are consistent with previous MMM campaigns and indicate that the screening campaign is feasible and useful to identify hypertension even in exceptional conditions such as the COVID-19 pandemic.

Prediction of unconventional magnetism in doped FeSb2.

It is commonly believed that the energy bands of typical collinear antiferromagnets (AFs), which have zero net magnetization, are Kramers spin-degenerate. Kramers nondegeneracy is usually associated with a global time-reversal symmetry breaking (e.g., via ferromagnetism) or with a combination of spin-orbit interaction and broken spatial inversion symmetry. Recently, another type of spin splitting was demonstrated to emerge in some collinear magnets that are fully spin compensated by symmetry, nonrelativistic, and not even necessarily noncentrosymmetric. These materials feature nonzero spin density staggered in real space as seen in traditional AFs but also spin splitting in momentum space, generally seen only in ferromagnets. This results in a combination of materials characteristics typical of both ferromagnets and AFs. Here, we discuss this recently discovered class with application to a well-known semiconductor, FeSb2, and predict that with certain alloying, it becomes magnetic and metallic and features the aforementioned magnetic dualism. The calculated energy bands split antisymmetrically with respect to spin-degenerate nodal surfaces rather than nodal points, as in the case of spin-orbit splitting. The combination of a large (0.2-eV) spin splitting, compensated net magnetization with metallic ground state, and a specific magnetic easy axis generates a large anomalous Hall conductivity (∼150 S/cm) and a sizable magnetooptical Kerr effect, all deemed to be hallmarks of nonzero net magnetization. We identify a large contribution to the anomalous response originating from the spin-orbit interaction gapped anti-Kramers nodal surfaces, a mechanism distinct from the nodal lines and Weyl points in ferromagnets.

Assessment of Purity, Functionality, Stability, and Lipid Composition of Cyclofos-nAChR-Detergent Complexes from Torpedo californica Using Lipid Matrix and Macroscopic Electrophysiology.

The main objective of the present study was to find detergents that can maintain the functionality and stability of the Torpedo californica nicotinic acetylcholine receptor (Tc-nAChR). We examined the functionality, stability, and purity analysis of affinity-purified Tc-nAChR solubilized in detergents from the Cyclofos (CF) family [cyclofoscholine 4 (CF-4), cyclofoscholine 6 (CF-6), and cyclofloscholine 7 (CF-7)]. The functionality of the CF-Tc-nAChR-detergent complex (DC) was evaluated using the Two Electrode Voltage Clamp (TEVC) method. To assess stability, we used the florescence recovery after photobleaching (FRAP) in Lipidic Cubic Phase (LCP) methodology. We also performed a lipidomic analysis using Ultra-Performance Liquid Chromatography (UPLC) coupled to electrospray ionization mass spectrometry (ESI-MS/MS) to evaluate the lipid composition of the CF-Tc-nAChR-DCs. The CF-4-Tc-nAChR-DC displayed a robust macroscopic current (- 200 ± 60 nA); however, the CF-6-Tc-nAChR-DC and CF-7-Tc-nAChR-DC displayed significant reductions in the macroscopic currents. The CF-6-Tc-nAChR and CF-4-Tc-nAChR displayed higher fractional florescence recovery. Addition of cholesterol produced a mild enhancement of the mobile fraction on the CF-6-Tc-nAChR. The lipidomic analysis revealed that the CF-7-Tc-nAChR-DC displayed substantial delipidation, consistent with the lack of stability and functional response of this complex. Although the CF-6-nAChR-DC complex retained the largest amount of lipids, it showed a loss of six lipid species [SM(d16:1/18:0); PC(18:2/14:1); PC(14:0/18:1); PC(16:0/18:1); PC(20:5/20:4), and PC(20:4/20:5)] that are present in the CF-4-nAChR-DC. Overall, the CF-4-nAChR displayed robust functionality, significant stability, and the best purity among the three CF detergents; therefore, CF-4 is a suitable candidate to prepare Tc-nAChR crystals for structural studies.

Chiral Magnons in Altermagnetic RuO_{2}.

Magnons in ferromagnets have one chirality, and typically are in the GHz range and have a quadratic dispersion near the zero wave vector. In contrast, magnons in antiferromagnets are commonly considered to have bands with both chiralities that are degenerate across the entire Brillouin zone, and to be in the THz range and to have a linear dispersion near the center of the Brillouin zone. Here we theoretically demonstrate a new class of magnons on a prototypical d-wave altermagnet RuO_{2} with the compensated antiparallel magnetic order in the ground state. Based on density-functional-theory calculations we observe that the THz-range magnon bands in RuO_{2} have an alternating chirality splitting, similar to the alternating spin splitting of the electronic bands, and a linear magnon dispersion near the zero wave vector. We also show that, overall, the Landau damping of this metallic altermagnet is suppressed due to the spin-split electronic structure, as compared to an artificial antiferromagnetic phase of the same RuO_{2} crystal with spin-degenerate electronic bands and chirality-degenerate magnon bands.

Cobalt-Catalyzed Decarboxylative Allylations: Development and Mechanistic Studies.

In recent years, there has been a concerted drive to develop methods that are greener and more sustainable. Being an earth-abundant transition metal, cobalt offers an attractive substitute for commonly employed precious metal catalysts, though reactions engaging cobalt are still less developed. Herein, we report a method to achieve the decarboxylative allylation of nitrophenyl alkanes, nitroalkanes, and ketones employing cobalt. The reaction allows for the formation of various substituted allylated products in moderate-excellent yields with a broad scope. Additionally, the synthetic potential of the methodology is demonstrated by the transformation of products into versatile heterocyclic motifs. Mechanistic studies revealed an in situ activation of the Co(II)/dppBz precatalyst by the carboxylate salt to generate a Co(I)-species, which is presumed to be the active catalyst.

Direct Aroylation of Olefins through a Cobalt/Photoredox-Catalyzed Decarboxylative and Dehydrogenative Coupling with α-Oxo Acids.

A photoredox/cobalt dual catalytic procedure has been developed that allows benzoylation of olefins. Here the photoredox catalyst effects the decarboxylation of α-ketoacids to form benzoyl radicals. After addition of this radical to styrenes, the cobalt catalyst abstracts a H-atom. Hydrogen evolution from the putative cobalt hydride intermediate allows a Heck-like aroylation without the need for a stoichiometric oxidant. Mechanistic studies reveal that electronically different styrenes lead to a curved Hammett plot, thus suggesting a change in product-determining step in the catalytic mechanism.

Direct Aroylation of Olefins through a Cobalt/Photoredox-Catalyzed Decarboxylative and Dehydrogenative Coupling with α-Oxo Acids.

Invited for the cover of this issue is the group of Jon Tunge at the University of Kansas. The image depicts the direct cross-coupling of α-oxo acids and styrenes through the release of CO2 and H2 gas in a metallaphotoredox process. Read the full text of the article at 10.1002/chem.202202781.

Assessing the effect of non-pharmaceutical interventions on COVID-19 transmission in Spain, 30 August 2020 to 31 January 2021.

BackgroundAfter a national lockdown during the first wave of the COVID-19 pandemic in Spain, regional governments implemented different non-pharmaceutical interventions (NPIs) during the second wave.AimTo analyse which implemented NPIs significantly impacted effective reproduction number (Rt) in seven Spanish provinces during 30 August 2020-31 January 2021.MethodsWe coded each NPI and levels of stringency with a 'severity index' (SI) and computed a global SI (mean of SIs per six included interventions). We performed a Bayesian change point analysis on the Rt curve of each province to identify possible associations with global SI variations. We fitted and compared several generalised additive models using multimodel inference, to quantify the statistical effect on Rt of the global SI (stringency) and the individual SIs (separate effect of NPIs).ResultsThe global SI had a significant lowering effect on the Rt (mean: 0.16 ± 0.05 units for full stringency). Mandatory closing times for non-essential businesses, limited gatherings, and restricted outdoors seating capacities (negative) as well as curfews (positive) were the only NPIs with a significant effect. Regional mobility restrictions and limited indoors seating capacity showed no effect. Our results were consistent with a 1- to 3-week-delayed Rt as a response variable.ConclusionWhile response measures implemented during the second COVID-19 wave contributed substantially to a decreased reproduction number, the effectiveness of measures varied considerably. Our findings should be considered for future interventions, as social and economic consequences could be minimised by considering only measures proven effective.

Lossless Medical Image Compression by Using Difference Transform.

This paper introduces a new method of compressing digital images by using the Difference Transform applied in medical imaging. The Difference Transform algorithm performs the decorrelation process of image data, and in this way improves the encoding process, achieving a file with a smaller size than the original. The proposed method proves to be competitive and in many cases better than the standards used for medical images such as TIFF or PNG. In addition, the Difference Transform can replace other transforms like Cosine or Wavelet.

Efficient Electrical Spin Splitter Based on Nonrelativistic Collinear Antiferromagnetism.

Spin-current generation by electrical means is among the core phenomena driving the field of spintronics. Using ab initio calculations we show that a room-temperature metallic collinear antiferromagnet RuO_{2} allows for highly efficient spin-current generation, arising from anisotropically spin-split bands with conserved up and down spins along the Néel vector axis. The zero net moment antiferromagnet acts as an electrical spin splitter with a 34° propagation angle between spin-up and spin-down currents. The corresponding spin conductivity is a factor of 3 larger than the record value from a survey of 20 000 nonmagnetic spin-Hall materials. We propose a versatile spin-splitter-torque concept circumventing limitations of spin-transfer and spin-orbit torques in present magnetic memory devices.

May Measurement Month 2019: an analysis of blood pressure screening results from Venezuela, Latin America.

Cardiovascular diseases are the main cause of death in Venezuela. Raised blood pressure (BP) accompanied by diabetes mellitus, obesity, lipid abnormalities, and tobacco usage are the biggest contributors to mortality. The May Measurement Month (MMM) campaign is a global initiative aimed to raising awareness of hypertension, which has been conducted in Venezuela since 2017. MMM2019 included 24 672 subjects (mean age: 54.7 years, SD 25.2, 63.1% female). The proportion with hypertension was 48.9%; 14.3% were unknown hypertensives, 35.5% of those who receiving treatment had uncontrolled hypertension (systolic BP ≥140 mmHg and/or diastolic BP ≥90 mmHg); when considering all hypertensives, 53.3% were controlled. Sixty per cent of those on anti-hypertensive medication were on monotherapy, 27.7% were on two, and 7.7% were on three or more drugs. Body mass index, calculated for the total population, was on average 25.6 (SD: 4.8) kg/m2. 16.2% of participants were classified as obese, 34.0% as overweight, and 4.0% were classified as underweight. Diabetes mellitus was reported by 9.4%, smoking by 7.3%, and 10.5% reported drinking alcohol regularly. Conditions associated with higher BP levels were obesity, diabetes mellitus, and women with a history of hypertension during a previous pregnancy. These results are consistent with the two previous MMM campaigns and indicate that repeated screening can routinely identify hypertension. There is an urgent need for Venezuela to implement programmes of detection, treatment, and control not only for hypertension but also for other common cardiovascular risk factors.

Severe COVID-19 initially presenting as mesenteric adenopathy.

Coronavirus disease 2019 (COVID-19) can present with abdominal pain in children and adults. Most imaging findings have been limited to characteristic lung findings, as well as one report of bowel-ischemia-related findings in adults. We report a case of COVID-19 in a healthy teenager who initially presented with isolated mesenteric adenopathy, typically a self-limited illness, which progressed to severe illness requiring intensive care before complete recovery. The boy tested negative for COVID-19 twice by polymerase chain reaction (PCR) from upper respiratory swabs before sputum PCR resulted positive. A high index of suspicion should be maintained for COVID-19 given the continued emergence of new manifestations of the disease.

Experiences of early graduate medical students working in New York hospitals during the COVID-19 pandemic: a mixed methods study.

BACKGROUND: The coronavirus disease 2019 (COVID-19) pandemic presented the world with a sudden need for additional medical professionals. Senior medical students were identified as potential workers and many worldwide graduated early to serve as Junior Physicians in hospitals. The authors sought to identify factors that informed the decision to work, describe experiences in this capacity, and elucidate benefits for trainees. METHODS: The investigators conducted a mixed-methods observational cohort study of early medical graduates eligible to work as Junior Physicians at two New York medical centers in April/May 2020 during an initial surge in COVID-19 hospitalizations. Graduates were surveyed, and a sample of Junior Physicians participated in a focus group. Survey responses of those who worked were compared to those who did not. Focus group responses were transcribed, coded, and thematically analyzed. RESULTS: Fifty-nine graduates completed the study methods and 39 worked as Junior Physicians. Primary reasons for working included duty to help (39 [100%]), financial incentive (32 [82%]), desire to learn about pandemic response (25 [64%]), and educational incentive (24 [62%]). All had direct contact with COVID-19 patients, believed working was beneficial to their medical training, and were glad they worked. None contracted a symptomatic infection while working. Compared with non-Junior Physicians, Junior Physicians reported increased comfort levels in completing medical intern-level actions like transitions of care functions, such as writing transfer notes (P < 0.01), writing discharge orders (P = 0.01), and providing verbal sign out (P = 0.05), and they reported more comfort in managing COVID-19 patients. Sixteen themes emerged from the focus group and were placed into four categories: development of skills, patient care, safety, and wellness. CONCLUSIONS: Senior medical students chose to work as Junior Physicians for both personal and educational reasons. Experiences were beneficial to trainees and can inform future innovations in medical education.

Pursuing High-Resolution Structures of Nicotinic Acetylcholine Receptors: Lessons Learned from Five Decades.

Since their discovery, nicotinic acetylcholine receptors (nAChRs) have been extensively studied to understand their function, as well as the consequence of alterations leading to disease states. Importantly, these receptors represent pharmacological targets to treat a number of neurological and neurodegenerative disorders. Nevertheless, their therapeutic value has been limited by the absence of high-resolution structures that allow for the design of more specific and effective drugs. This article offers a comprehensive review of five decades of research pursuing high-resolution structures of nAChRs. We provide a historical perspective, from initial structural studies to the most recent X-ray and cryogenic electron microscopy (Cryo-EM) nAChR structures. We also discuss the most relevant structural features that emerged from these studies, as well as perspectives in the field.

Sequential purification and characterization of Torpedo californica nAChR-DC supplemented with CHS for high-resolution crystallization studies.

Over the past 10 years we have been developing a multi-attribute analytical platform that allows for the preparation of milligram amounts of functional, high-pure, and stable Torpedo (muscle-type) nAChR detergent complexes for crystallization purpose. In the present work, we have been able to significantly improve and optimize the purity and yield of nicotinic acetylcholine receptors in detergent complexes (nAChR-DC) without compromising stability and functionality. We implemented new methods in the process, such as analysis and rapid production of samples for future crystallization preparations. Native nAChR was extracted from the electric organ of Torpedo californica using the lipid-like detergent LysoFos Choline 16 (LFC-16), followed by three consecutive steps of chromatography purification. We evaluated the effect of cholesteryl hemisuccinate (CHS) supplementation during the affinity purification steps of nAChR-LFC-16 in terms of receptor secondary structure, stability and functionality. CHS produced significant changes in the degree of ß-secondary structure, these changes compromise the diffusion of the nAChR-LFC-16 in lipid cubic phase. The behavior was reversed by Methyl-ß-Cyclodextrin treatment. Also, CHS decreased acetylcholine evoked currents of Xenopus leavis oocyte injected with nAChR-LFC-16 in a concentration-dependent manner. Methyl-ß-Cyclodextrin treatment do not reverse functionality, however column delipidation produced a functional protein similar to nAChR-LFC-16 without CHS treatment.

Partial or complete? The evolution of post-juvenile moult strategies in passerine birds.

Moulting strategies in birds have evolved to avoid overlap with, or prepare for, other demanding parts of the annual cycle, such as reproduction or migration. When moulting for the first time after leaving the nest, young birds replace their typically poor-quality plumage during the post-juvenile moult. The extent of this moult varies between species from partial to complete. Earlier studies, restricted to Western Palearctic birds, suggest that in most species a complete post-juvenile moult may not be possible simply because young birds are constrained by not having the same access to resources as adults, unless environmental conditions are favourable. These studies also show that complete post-juvenile moult is more common in species with poor-quality nest-grown plumage. We expanded the spatial and taxonomic scope of previous studies to 1,315 species of passerines from across the world and considered both the role of constraints, plumage quality and other selective pressures favouring a complete post-juvenile moult. Thus, we test whether complete moult is more prevalent in species where nest-grown feathers are presumably of poor quality (shorter nestling period), that live in environments that foster quick plumage degradation (open habitats, high insolation and humidity), and where males are under strong sexual selection. Our data reveal that 24% of species carry out a complete post-juvenile moult, and that this trait has a strong phylogenetic signal. Complete moult is more common in species that inhabit warmer regions and open habitats, show no delayed plumage maturation and have higher levels of sexual dichromatism (indicative of strong sexual selection). Neither the presumed quality of the nest-grown plumage nor living in regions with high insolation correlates with complete moult. In conclusion, the evolution of complete post-juvenile moult not only depends on whether birds can perform a complete moult (i.e. suitable environmental conditions) but also on the strength of selection associated with the need of a complete moult. In particular, the necessity to keep the plumage in good condition in challenging environments and the benefits associated with producing adult-like plumage colours to attract mates or deter rivals seem to play an important role.

The fingerprint of essential bio-oils by Fourier transform ion cyclotron resonance mass spectrometry.

Six essential oils were analyzed by Fourier transform ion cyclotron resonance mass spectrometry coupled to negative-ion electrospray ionization (ESI(-)/FT-ICR MS). ESI offers selective ionization of a compound's polar functional groups containing nitrogen and oxygen heteroatoms. ESI in negative-ion mode allows the identification of the acidic compounds. The results showed that the samples contain between 1100-3600 individual molecular compositions, which corresponds to the greatest number of species detected to date in essential oils obtained from aromatic plant material. The compositions cover a mass range between m/z 150-500 with up to 41 carbon atoms. The dominant organic constituents of the essential oils correspond to species incorporating 2-5 oxygen atoms, detected as deprotonated/sodiated/chlorinated species. A set of 580 molecular assignments were found in common across all the samples and for the first time, a set of unique molecular systems were identified, and up to 1373 species as a unique composition for each essential oil. The molecular distributions plotted in van Krevelen diagrams (classified by their H/C vs. O/C values) suggest the presence of species with long alkyl chains and low numbers of rings plus double bonds.

Mycobacteria manipulate macrophage recruitment through coordinated use of membrane lipids.

The evolutionary survival of Mycobacterium tuberculosis, the cause of human tuberculosis, depends on its ability to invade the host, replicate, and transmit infection. At its initial peripheral infection site in the distal lung airways, M. tuberculosis infects macrophages, which transport it to deeper tissues. How mycobacteria survive in these broadly microbicidal cells is an important question. Here we show in mice and zebrafish that M. tuberculosis, and its close pathogenic relative Mycobacterium marinum, preferentially recruit and infect permissive macrophages while evading microbicidal ones. This immune evasion is accomplished by using cell-surface-associated phthiocerol dimycoceroserate (PDIM) lipids to mask underlying pathogen-associated molecular patterns (PAMPs). In the absence of PDIM, these PAMPs signal a Toll-like receptor (TLR)-dependent recruitment of macrophages that produce microbicidal reactive nitrogen species. Concordantly, the related phenolic glycolipids (PGLs) promote the recruitment of permissive macrophages through a host chemokine receptor 2 (CCR2)-mediated pathway. Thus, we have identified coordinated roles for PDIM, known to be essential for mycobacterial virulence, and PGL, which (along with CCR2) is known to be associated with human tuberculosis. Our findings also suggest an explanation for the longstanding observation that M. tuberculosis initiates infection in the relatively sterile environment of the lower respiratory tract, rather than in the upper respiratory tract, where resident microflora and inhaled environmental microbes may continually recruit microbicidal macrophages through TLR-dependent signalling.