COVID-19 policy response: Perspectives of key stakeholders in Mexico's health system and implications for resilience.

Purpose: This study aims to analyze the perspectives of key stakeholders within Mexico's health system regarding the policy response to COVID-19 in 2020 to inform future policy formulation.

Methods: This qualitative study is based on a secondary data analysis (SDA) derived from a broader primary research on governance in Mexico during the COVID-19 pandemic. The analysis adopted a grounded theory approach to examine 15 semi-structured interviews with prominent figures in the health system between June and November 2020.

Lessons learned during the journey of data: from experiment to model for predicting kinase affinity, selectivity, polypharmacology, and resistance.

Recent advances in machine learning (ML) are reshaping drug discovery. Structure-based ML methods use physically-inspired models to predict binding affinities from protein:ligand complexes. These methods promise to enable the integration of data for many related targets, which addresses issues related to data scarcity for single targets and could enable generalizable predictions for a broad range of targets, including mutants.

Mechanistic insights into G-protein coupling with an agonist-bound G-protein-coupled receptor.

G-protein-coupled receptors (GPCRs) activate heterotrimeric G proteins by promoting guanine nucleotide exchange. Here, we investigate the coupling of G proteins with GPCRs and describe the events that ultimately lead to the ejection of GDP from its binding pocket in the Gα subunit, the rate-limiting step during G-protein activation. Using molecular dynamics simulations, we investigate the temporal progression of structural rearrangements of GDP-bound G protein (G·GDP; hereafter G) upon coupling to the β-adrenergic receptor (βAR) in atomic detail.

Cyclical heat stress during lactation influences the microstructure of the bovine mammary gland.

This study aimed to evaluate the effect of heat stress on mammary epithelial cell (MEC) losses into milk, secretory mammary tissue structure, and mammary epithelial cell activity. Sixteen multiparous Holstein cows (632 ± 12 kg BW) approximately 100 DIM housed in climate-controlled rooms were paired by BW and randomly allocated to one of 2 treatments, heat stress (HS) or pair-feeding thermoneutral (PFTN) using 2 cohorts. Each cohort was subjected to 2 periods of 4 d each.

Time-resolved cryo-EM of G-protein activation by a GPCR.

G-protein-coupled receptors (GPCRs) activate heterotrimeric G proteins by stimulating guanine nucleotide exchange in the Gα subunit. To visualize this mechanism, we developed a time-resolved cryo-EM approach that examines the progression of ensembles of pre-steady-state intermediates of a GPCR-G-protein complex. By monitoring the transitions of the stimulatory G protein in complex with the β-adrenergic receptor at short sequential time points after GTP addition, we identified the conformational trajectory underlying G-protein activation and functional dissociation from the receptor.

Relationships between gastrointestinal permeability, heat stress, and milk production in lactating dairy cows.

Heat stress (HS) is a global issue that decreases farm profits and compromises animal welfare. To distinguish between the direct and indirect effects of HS, 16 multiparous Holstein cows approximately 100 DIM were assigned to one of 2 treatments: pair fed to match HS cow intake, housed in thermoneutral conditions (PFTN, n = 8) or cyclical HS (n = 8). All cows were subjected to 2 experimental periods.

Inhibitory to non-inhibitory evolution of the subunit of the FF-ATPase of and -proteobacteria as related to mitochondrial endosymbiosis.

The subunit is a potent inhibitor of the FF-ATPase of (PdFF-ATPase) and related -proteobacteria different from the other two canonical inhibitors of bacterial () and mitochondrial (IF) FF-ATPases. mimics mitochondrial IF in its inhibitory N-terminus, blocking the PdFF-ATPase activity as a unidirectional pawl-ratchet and allowing the PdFF-ATP synthase turnover. is essential for the respiratory growth of , as we showed by a knockout.

Milk production and anatomical udder capacity changes of udder halves subjected to increased milking frequency at two stages of lactation.

This study aimed to characterize the effects of increased milking frequency (IMF) at early and mid-lactation on milk yield and its association with changes in cistern and alveolar capacity. Fourteen multiparous Holstein cows were subjected to IMF using the unilateral frequent milking method from 3 to 24 d in milk (DIM). At mid-lactation, cows were randomly assigned to 1 of 2 treatments: control or repeated.

Digital tomosynthesis improves chest radiograph accuracy and reduces microbiological false negatives in COVID-19 diagnosis.

Purpose: Diagnosing pneumonia by radiograph is improvable. We aimed (a) to compare radiograph and digital thoracic tomosynthesis (DTT) performances and agreement for COVID-19 pneumonia diagnosis, and (b) to assess the DTT ability for COVID-19 diagnosis when polymerase chain reaction (PCR) and radiograph are negative.

Methods: Two emergency radiologists with 11 (ER1) and 14 experience-years (ER2) retrospectively evaluated radiograph and DTT images acquired simultaneously in consecutively clinically suspected COVID-19 pneumonia patients in March 2020-January 2021.

Ordered-domain unfolding of thermophilic isolated β subunit ATP synthase.

The flexibility of the ATP synthase's β subunit promotes its role in the ATP synthase rotational mechanism, but its domains stability remains unknown. A reversible thermal unfolding of the isolated β subunit (Tβ) of the ATP synthase from Bacillus thermophilus PS3, tracked through circular dichroism and molecular dynamics, indicated that Tβ shape transits from an ellipsoid to a molten globule through an ordered unfolding of its domains, preserving the β-sheet residual structure at high temperature. We determined that part of the stability origin of Tβ is due to a transversal hydrophobic array that crosses the β-barrel formed at the N-terminal domain and the Rossman fold of the nucleotide-binding domain (NBD), while the helix bundle of the C-terminal domain is the less stable due to the lack of hydrophobic residues, and thus the more flexible to trigger the rotational mechanism of the ATP synthase.

Function and dynamics of the intrinsically disordered carboxyl terminus of β2 adrenergic receptor.

Advances in structural biology have provided important mechanistic insights into signaling by the transmembrane core of G-protein coupled receptors (GPCRs); however, much less is known about intrinsically disordered regions such as the carboxyl terminus (CT), which is highly flexible and not visible in GPCR structures. The β adrenergic receptor's (βAR) 71 amino acid CT is a substrate for GPCR kinases and binds β-arrestins to regulate signaling. Here we show that the βAR CT directly inhibits basal and agonist-stimulated signaling in cell lines lacking β-arrestins.

Time-resolved cryo-EM of G protein activation by a GPCR.

G protein-coupled receptors (GPCRs) activate heterotrimeric G proteins by stimulating the exchange of guanine nucleotide in the Gα subunit. To visualize this mechanism, we developed a time-resolved cryo-EM approach that examines the progression of ensembles of pre-steady-state intermediates of a GPCR-G protein complex. Using variability analysis to monitor the transitions of the stimulatory Gs protein in complex with the β -adrenergic receptor (β AR) at short sequential time points after GTP addition, we identified the conformational trajectory underlying G protein activation and functional dissociation from the receptor.

[Pillars and lines of action for integrated and people- and community-centered health systemsPilares e linhas de ação para sistemas integrados de saúde centrados nas pessoas e nas comunidades].

This paper presents the position of the Latin American working group of the International Foundation for Integrated Care (IFIC). The working group brings together various Latin American actors and organizations in support of actions that facilitate the transformation of health systems in the region towards integrated systems that focus on people not as isolated individuals but as subjects of law in the complex social and environmental contexts where they live and interact. The working group proposes nine pillars of integrated care to be used as a conceptual framework for policy development and changes in practices: 1) shared vision and values; 2) population health; 3) people and communities as partners; 4) resilient communities; 5) capacities of human resources for health; 6) governance and leadership; 7) digital solutions; 8) aligned payment systems; and 9) public transparency.

Transcriptomic and Drug Discovery Analyses Reveal Natural Compounds Targeting the KDM4 Subfamily as Promising Adjuvant Treatments in Cancer.

KDM4 proteins are a subfamily of histone demethylases that target the trimethylation of lysines 9 and 36 of histone H3, which are associated with transcriptional repression and elongation respectively. Their deregulation in cancer may lead to chromatin structure alteration and transcriptional defects that could promote malignancy. Despite that KDM4 proteins are promising drug targets in cancer therapy, only a few drugs have been described as inhibitors of these enzymes, while studies on natural compounds as possible inhibitors are still needed.

Tethered agonist exposure in intact adhesion/class B2 GPCRs through intrinsic structural flexibility of the GAIN domain.

Adhesion G protein-coupled receptors (aGPCRs)/family B2 GPCRs execute critical tasks during development and the operation of organs, and their genetic lesions are associated with human disorders, including cancers. Exceptional structural aGPCR features are the presence of a tethered agonist (TA) concealed within a GPCR autoproteolysis-inducing (GAIN) domain and their non-covalent heteromeric two-subunit layout. How the TA is poised for activation while maintaining this delicate receptor architecture is central to conflicting signaling paradigms that either involve or exclude aGPCR heterodimer separation.

Radiology departments as COVID-19 entry-door might improve healthcare efficacy and efficiency, and emergency department safety.

Background: Possible COVID-19 pneumonia patients (ppCOVID-19) generally overwhelmed emergency departments (EDs) during the first COVID-19 wave. Home-confinement and primary-care phone follow-up was the first-level regional policy for preventing EDs to collapse. But when X-rays were needed, the traditional outpatient workflow at the radiology department was inefficient and potential interpersonal infections were of concern.

Energetic metabolism, milk production, and inflammatory response of transition dairy cows fed rumen-protected glucose.

Objectives were to evaluate the effects of rumen-protected glucose (RPG) supplementation on milk production, post-absorptive metabolism, and inflammatory biomarkers in transition dairy cows. Fifty-two multiparous cows were blocked by previous 305-d mature-equivalent milk (305ME) yield and randomly assigned to 1 of 2 iso-energetic and iso-nitrogenous treatments: (1) control diet (CON; n = 26) or (2) a diet containing RPG (pre-fresh 5.3% of dry matter and 6.

On the Thermal Stability of O-Methylguanine-DNA Methyltransferase from Archaeon by Molecular Dynamics Simulations.

We have employed molecular dynamics simulations to analyze the thermal stability of the O-methylguanine-DNA methyltransferase (MGMT) protein, both hyperthermophilic archaeon (Pk-MGMT) and its mesophilic homologue pair, obtained from enterobacterium (AdaC). This theoretical study was done at three different temperatures: 302, 371, and 450 K. The molecular dynamics has been performed in explicit aqueous solvent during a period of time of 95 ns, including periodic boundary conditions and constant pressure.

Nanostructured oleic acid/polysorbate 80 emulsions with diminished toxicity in NL-20 cell line: Insights of potential drug carriers.

Nanoemulsions (NE) are nowadays required drug nanocarriers. We have selected i) oleic acid (OA) as oil (O), ii) polysorbate 80 (PS) as surfactant (S), and iii) water (W) in a prototype NE. Our best formulation had O:S ratio [OA]/[PS] = 0.

Influence of physicochemical factors on environmental availability and distribution of semiochemicals that affect and phylogenetically close organisms: classification by VHWOC PCA-clustering.

parasites larvae following the interception of the semiochemicals involved in bee communication; thus, the semiochemical availability and distribution pathways take place within different physicochemical environments. The structure of 172 molecules with semiochemical activity on was used to compute the representative physicochemical descriptors of the thermodynamic partition among different physicochemical environments: vapor pressure (V), Henry's coefficient (H), water solubility constant (W), octanol-water partition coefficient (O) and organic carbon partition coefficient (C); VHWOC. The principal component analysis (PCA) and hierarchical clustering of VHWOC descriptors allowed us to establish the trend in availability and distribution of the semiochemicals resulting in a 4 classes model of physicochemical environments: Class 1, Soluble/Volatile; Class 2, Soluble; Class 3, Contact; Class 4, Adsorbed/Volatile.

Physicochemical characterization and thermal behavior of hexosomes containing ketoconazole as potential topical antifungal delivery system.

Objective: The main purpose of this article is to show the valuable characteristics that liotropic liquid crystal systems possess to be employed as new drug delivery systems.

Significance: Colloidal aqueous dispersions of lyotropic liquid crystal mesophases such as the identified as cubosomes and hexosomes, and so on, have received considerable attention due to their unique nanostructures and their thermodynamic properties, which provide the potential as a sustained drug release matrix. Additionally, their large surface area and similarity with the liquid crystal structures of intercellular lipids of stratum corneum enhances the interaction with the skin and mucous, increasing the potential for topical drug delivery efficiency of biopharmaceutical class II drugs as the antifungal ketoconazole.

Defining the architecture of KPC-2 Carbapenemase: identifying allosteric networks to fight antibiotics resistance.

The rise of multi-drug resistance in bacterial pathogens is one of the grand challenges facing medical science. A major concern is the speed of development of β-lactamase-mediated resistance in Gram-negative species, thus putting at risk the efficacy of the most recently approved antibiotics and inhibitors, including carbapenems and avibactam, respectively. New strategies to overcome resistance are urgently required, which will ultimately be facilitated by a deeper understanding of the mechanisms that regulate the function of β-lactamases such as the Klebsiella Pneumoniae carbapenemases (KPCs).

Exploring interfacial water trapping in protein-ligand complexes with multithermal titration calorimetry.

In this work, we examine the hypothesis about how trapped water molecules at the interface between triosephosphate isomerase (TIM) and either of two phosphorylated inhibitors, 2-phosphoglycolate (2PG) or phosphoglycolohydroxamate (PGH), can explain the anomalous highly negative binding heat capacities (ΔC) of both complexes, TIM-2PG and TIM-PGH. We performed fluorimetric titrations of the enzyme with PGH inhibitor under osmotic stress conditions, using various concentrations of either osmolyte: sucrose, ethylene glycol or glycine betaine. We also analyze the binding processes under various stressor concentrations using a novel calorimetric methodology that allows ΔC determinations in single experiments: Multithermal Titration Calorimetry.

Exploring the Dynamics of Propeller Loops in Human Telomeric DNA Quadruplexes Using Atomistic Simulations.

We have carried out a series of extended unbiased molecular dynamics (MD) simulations (up to 10 μs long, ∼162 μs in total) complemented by replica-exchange with the collective variable tempering (RECT) approach for several human telomeric DNA G-quadruplex (GQ) topologies with TTA propeller loops. We used different AMBER DNA force-field variants and also processed simulations by Markov State Model (MSM) analysis. The slow conformational transitions in the propeller loops took place on a scale of a few μs, emphasizing the need for long simulations in studies of GQ dynamics.

Hierarchical Time-Lagged Independent Component Analysis: Computing Slow Modes and Reaction Coordinates for Large Molecular Systems.

Analysis of molecular dynamics, for example using Markov models, often requires the identification of order parameters that are good indicators of the rare events, i.e. good reaction coordinates.