Resident macrophage-dependent immune cell scaffolds drive anti-bacterial defense in the peritoneal cavity.

Peritoneal immune cells reside unanchored within the peritoneal fluid in homeostasis. Here, we examined the mechanisms that control bacterial infection in the peritoneum using a mouse model of abdominal sepsis following intraperitoneal Escherichia coli infection. Whole-mount immunofluorescence and confocal microscopy of the peritoneal wall and omentum revealed that large peritoneal macrophages (LPMs) rapidly cleared bacteria and adhered to the mesothelium, forming multilayered cellular aggregates composed by sequentially recruited LPMs, B1 cells, neutrophils, and monocyte-derived cells (moCs). The formation of resident macrophage aggregates (resMφ-aggregates) required LPMs and thrombin-dependent fibrin polymerization. E. coli infection triggered LPM pyroptosis and release of inflammatory mediators. Resolution of these potentially inflammatory aggregates required LPM-mediated recruitment of moCs, which were essential for fibrinolysis-mediated resMφ-aggregate disaggregation and the prevention of peritoneal overt inflammation. Thus, resMφ-aggregates provide a physical scaffold that enables the efficient control of peritoneal infection, with implications for antimicrobial immunity in other body cavities, such as the pleural cavity or brain ventricles.

40S Ribosomal Protein S6 Kinase Integrates Daylength Perception and Growth Regulation in Arabidopsis thaliana.

Plant growth occurs via the interconnection of cell growth and proliferation in each organ following specific developmental and environmental cues. Therefore, different photoperiods result in distinct growth patterns due to the integration of light and circadian perception with specific Carbon (C) partitioning strategies. In addition, the TARGET OF RAPAMYCIN (TOR) kinase pathway is an ancestral signaling pathway that integrates nutrient information with translational control and growth regulation. Recent findings in Arabidopsis (Arabidopsis thaliana) have shown a mutual connection between the TOR pathway and the circadian clock. However, the mechanistical network underlying this interaction is mostly unknown. Here, we show that the conserved TOR target, the 40S ribosomal protein S6 kinase (S6 K) is under circadian and photoperiod regulation both at the transcriptional and post-translational level. Total S6 K (S6K1 and S6K2) and TOR-dependent phosphorylated-S6 K protein levels were higher during the light period and decreased at dusk especially under short day conditions. Using chemical and genetic approaches we found that the diel pattern of S6 K accumulation results from 26S proteasome-dependent degradation and is altered in mutants lacking the circadian F-box protein ZEITLUPE (ZTL), further strengthening our hypothesis that S6 K could incorporate metabolic signals via TOR, which are also under circadian regulation. Moreover, under short days when C/energy levels are limiting, changes in S6K1 protein levels affected starch, sucrose and glucose accumulation and consequently impacted root and rosette growth responses. In summary, we propose that S6K1 constitutes a missing molecular link where day-length perception, nutrient availability and TOR pathway activity converge to coordinate growth responses with environmental conditions.

Bioactive compounds: Application of albumin nanocarriers as delivery systems.

Enriched products with bioactive compounds (BCs) show the capacity to produce a wide range of possible health effects. Most BCs are essentially hydrophobic and sensitive to environmental factors; so, encapsulation becomes a strategy to solve these problems. Many globular proteins have the intrinsic ability to bind, protect, encapsulate, and introduce BCs into nutraceutical or pharmaceutical matrices. Among them, albumins as human serum albumin (HSA), bovine serum albumin (BSA), ovalbumin (OVA) and α-lactalbumin (ALA) are widely abundant, available, and applied in many industrial sectors, becoming promissory materials to encapsulate BCs. Therefore, this review focuses on researches about the main groups of natural origin BCs (namely phenolic compounds, lipids, vitamins, and carotenoids), the different types of nanostructures based on albumins to encapsulate them and the main fields of application for BCs-loaded albumin systems. In this context, phenolic compounds (catechins, quercetin, and chrysin) are the most extensively BCs studied and encapsulated in albumin-based nanocarriers. Other extensively studied subgroups are stilbenes and curcuminoids. Regarding lipids and vitamins; terpenes, carotenoids (ß-carotene), and xanthophylls (astaxanthin) are the most considered. The main application areas of BCs are related to their antitumor, anti-inflammatory, and antioxidant properties. Finally, BSA is the most used albumin to produced BCs-loaded nanocarriers.

Foaming properties are improved by interactions between brewer's spent grain proteins and carrageenans in aqueous solution.

BACKGROUND: Foaming properties and macromolecular interactions in solution among brewer's spent grain proteins (CP) and iota and lambda carrageenans (i-CG and l-CG, respectively) as a function of aqueous medium pH (2-6) and protein-polysaccharide ratio, RCP:CG (1:1, 2:1 and 4:1), were studied. At these conditions, the CP colloidal stability was favored by the formation of soluble electrostatic complexes with CG. Fluorescence (intrinsic and extrinsic) spectroscopy and dynamic light scattering techniques, including particle size and ζ-potential analysis, were applied to know the phase behavior of the biopolymer systems. The bubbling method was used to produce foams, and the foam expansion (%) and half-life time (t1/2 ) were determined. RESULTS: Both CG promoted an increased Trp fluorescence emission depending on the pH, suggesting conformational changes in CP. The CG in mixed systems produced a significant decrease in the extrinsic fluorescence intensity, mainly at low pH values, highlighting a reduction in CP surface hydrophobicity. At the examined pH range, the ζ-potential values for mixed-systems were negative, and their magnitudes were intermediate between CP and CG, revealing the associative electrostatic nature of biopolymer interactions, which were dependent on the RCP:CG . The particle size analysis confirmed the formation of soluble electrostatic complexes in solution. Finally, using i-CG at pH 2 or 3 and 2:1 RCP:CG , the best foaming properties for mixed systems were observed. CONCLUSION: The formation of electrostatic complexes with a compact assembly among biopolymers, high negative net charge, and colloidal stability convert the CP-CG mixed solutions into promising biopolymer systems for food foams production. © 2022 Society of Chemical Industry.

In vitro assessment of two novel Cellulases from Trabulsiella odontotermitis for agricultural waste utilization.

BACKGROUND: The production of agricultural wastes still growing as a consequence of the population growing. However, the majority of these residues are under-utilized due their chemical composition, which is mainly composed by cellulose. Actually, the search of cellulases with high efficiency to degrade this carbohydrate remains as the challenge. In the present experiment, two genes encoding an endoglucanase (EC 3.2.1.4) and ß-glucosidase (EC 3.2.1.21) were overexpressed in Escherichia coli and their recombinant enzymes (egl-FZYE and cel-FZYE, respectively) characterized. Those genes were found in Trabulsiella odontermitis which was isolated from the gut of termite Heterotermes sp. Additionally, the capability to release sugars from agricultural wastes was evaluated in both enzymes, alone and in combination. RESULTS: The results have shown that optimal pH was 6.0 and 6.5, reaching an activity of 1051.65 ± 47.78 and 607.80 ± 10.19 U/mg at 39 °C, for egl-FZYE and cel-FZYE, respectively. The Km and Vmax for egl-FZYE using CMC as substrate were 11.25 mg/mL and 3921.57 U/mg, respectively, whereas using Avicel were 15.39 mg/mL and 2314.81 U/mg, respectively. The Km and Vmax for cel-FZYE using Avicel as substrate were 11.49 mg/mL and 2105.26 U/mg, respectively, whereas using CMC the enzyme did not had activity. Both enzymes had effect on agricultural wastes, and their effect was improved when they were combined reaching an activity of 955.1 ± 116.1, 4016.8 ± 332 and 1124.2 ± 241 U/mg on corn stover, sorghum stover and pine sawdust, respectively. CONCLUSIONS: Both enzymes were capable of degrading agricultural wastes, and their effectiveness was improved up to 60% of glucose released when combined. In summary, the results of the study demonstrate that the recombinant enzymes exhibit characteristics that indicate their value as potential feed additives and that the enzymes could be used to enhance the degradation of cellulose in the poor-quality forage generally used in ruminant feedstuffs.

Anatomical variations of the sacro-iliac joint: a computed tomography study.

PURPOSE: To identify, describe, and compare the prevalence of sacroiliac joint anatomical variants. METHODS: A retrospective study was performed on computed tomography scans. Joint space was measured, and variants were classified as accessory joint, ileosacral complex, bipartite bony plate, semicircular defect, iliac bony plate, and ossification centers. RESULTS: 400 scans were analyzed. Mean age was 49 years, 180 men (45%), and 220 women (55%). 209 (52.2%) patients presented an anatomical variant with higher prevalence in women (65.4% vs 36.2%), and those older than 40 years of age (60% vs 40%). Mean joint space was similar bilaterally (right 2.41 ± 0.65 mm vs. left 2.37 ± 0.65 mm). Prevalence was: 19.8% accessory joint, 6.5% ileosacral complex, 12.3% bipartite bony plate, 8% semicircular defect, 5% iliac bony plate, and 0.8% for ossification centers. CONCLUSION: Prevalence of anatomical variants of the SIJ is higher in the Hispanic population, women, and those older than 40 years.

Serial-section atlas of the Tritonia pedal ganglion.

The pedal ganglion of the nudibranch gastropod Tritonia diomedea has been the focus of neurophysiological studies for more than 50 yr. These investigations have examined the neural basis of behaviors as diverse as swimming, crawling, reflex withdrawals, orientation to water flow, orientation to the earth's magnetic field, and learning. Despite this sustained research focus, most studies have confined themselves to the layer of neurons that are visible on the ganglion surface, leaving many neurons, which reside in deeper layers, largely unknown and thus unstudied. To facilitate work on such neurons, the present study used serial-section light microscopy to generate a detailed pictorial atlas of the pedal ganglion. One pedal ganglion was sectioned horizontally at 2-µm intervals and another vertically at 5-µm intervals. The resulting images were examined separately or combined into stacks to generate movie tours through the ganglion. These were also used to generate 3D reconstructions of individual neurons and rotating movies of digitally desheathed whole ganglia to reveal all surface neurons. A complete neuron count of the horizontally sectioned ganglion yielded 1,885 neurons. Real and virtual sections from the image stacks were used to reveal the morphology of individual neurons, as well as the major axon bundles traveling within the ganglion to and between its several nerves and connectives. Extensive supplemental data are provided, as well as a link to the Dryad Data Repository site, where the complete sets of high-resolution serial-section images can be downloaded. NEW & NOTEWORTHY Because of the large size and relatively low numbers of their neurons, gastropod mollusks are widely used for investigations of the neural basis of behavior. Most studies, however, focus on the neurons visible on the ganglion surface, leaving the majority, located out of sight below the surface, unexamined. The present light microscopy study generates the first detailed visual atlas of all neurons of the highly studied Tritonia pedal ganglion.

Controlling Foam Stability with the Ratio of Myristic Acid to Choline Hydroxide.

The interfacial and foam properties of a model system based on the mixture between myristic acid and choline hydroxide have been investigated as a function of the molar ratio ( R) between these two components and temperature. The aim of this study was to obtain insight on the links between the self-assemblies in bulk and in the foam liquid channels, the surfactant packing at the interface, and the resulting foam properties and stability. A multiscale approach was used combining small angle neutron scattering, specular neutron reflectivity, surface tension measurements, and photography. We highlighted three regimes of foam stability in this system by modifying R: high foam stability for R < 1, intermediate at R ∼ 1, and low for R > 1. The different regimes come from the pH variations in bulk linked to R. The pH plays a crucial role at the molecular scale by setting the ionization state of the myristic acid molecules adsorbed at the gas-liquid interface, which in turn controls both the properties of the monolayer and the stability of the films separating the bubbles. The main requirement to obtain stable foams is to set the pH close to the p Ka in order to have a mixture of protonated and ionized molecules giving rise to intermolecular hydrogen bonds. As a result, a dense monolayer is formed at the interface with a low surface tension. R also modifies the structure of self-assembly in bulk and therefore within the foam, but such a morphological change has only a minor effect on the foam stability. This study confirms that foam stability in surfactant systems having a carboxylic acid as polar headgroup is mainly linked to the ionization state of the molecules at the interface.

Application of a MEMS-Based TRNG in a Chaotic Stream Cipher.

In this work, we used a sensor-based True Random Number Generator in order to generate keys for a stream cipher based on a recently published hybrid algorithm mixing Skew Tent Map and a Linear Feedback Shift Register. The stream cipher was implemented and tested in a Field Programmable Gate Array (FPGA) and was able to generate 8-bit width data streams at a clock frequency of 134 MHz, which is fast enough for Gigabit Ethernet applications. An exhaustive cryptanalysis was completed, allowing us to conclude that the system is secure. The stream cipher was compared with other chaotic stream ciphers implemented on similar platforms in terms of area, power consumption, and throughput.

Risk factors for amputation in periprosthetic knee infection.

Treatment for prosthetic knee replacement is becoming more common. Infection is an arthroplasty-related complication leading to prolonged hospitalization, multiple surgical procedures, permanent loss of the implant, impaired function, impaired quality of life and even amputation of the limb. Previous studies have evaluated the risk factors associated with periprosthetic knee infection, but scarce information related to risk factors associated with amputation in this group of patients is available. The purpose of this study was to identify risk factors for amputation in periprosthetic infected knee through a case-control study, analyzing patients treated from January 2012 to November 2016 in a hospital with a high incidence of this diagnosis. We included 183 patients with periprosthetic knee infection; 23 required amputation as definitive management (cases). We found that patients with surgical time >120 min (p = 0.01), surgical risk higher than two points according to the American Society of Anesthesiology score (p = 0.00), smokers (p = 0.04), obesity and diabetes mellitus (p = 0.00) had an increased risk of amputation.

Development, characterization, and evaluation of the antioxidant activity of nanocarriers based on surfactant swollen micelles that encapsulate essential oils.

This work aimed to compare the performance of two relatively underexplored methods for the swollen micelles (SMs) production as nanocarriers for essential oils (EOs). Origanum vulgare and Thymus vulgaris EOs were examined. The first method (SMs-1), involved a self-assembly process, while the second one (SMs-2), employed titration operation of an emulsion into a surfactant solution for SMs formation. Tween 80 and ethanol were used as surfactant and co-surfactant, respectively. The solubilization kinetics and the saturation concentration of EOs were determined. Particle size (measured by DLS) and encapsulation efficiency (EE) were the control parameters assessed, along with the EOs-loaded SMs' stability during 30 days of storage. Additionally, the EOs-loaded SMs' morphology was analyzed using atomic force microscopy (AFM). Finally, the antioxidant activity through the ABTS+ radical scavenging and the reducing power of EOs encapsulated in SMs was determined. The results showed that the solubilization of EOs in SMs was a rapid process with high EE. EOs-loaded SMs-2 systems exhibited greater colloidal stability and higher EE compared to EOs-loaded SMs-1 systems, showing smaller and more homogeneous particle sizes. Moreover, EOs-loaded SMs-2 systems maintained constant EE throughout the storage period. AFM imaging confirmed the rounded and heterogeneous morphology of EOs-loaded SMs-1 and the smaller, more homogeneous, and spherical morphology of EOs-loaded SMs-2. EOs-loaded SMs-2 showed high ABTS+ radical scavenging and reducing power when encapsulated in SMs. In conclusion, the SMs-2 method emerged as an effective approach for producing efficient nanocarriers for EOs, signifying a promising path for future developments in antioxidant delivery systems.

Interface fails and Young's module approximation of multilayer flexible devices through finite element method.

Electronic flexible devices are prone to degrade their electrical performance or lose functionality when subjected to deformations. Brittle fracture is a common damaging effect observed in devices composed of low-thickness layered materials stacked onto a flexible substrate by dissimilar mechanical properties interaction. This work studies the mechanical behavior of Organic Flexible Solar Cells (OFSC) with a heterostructure PET/ITO/P3HT:PCBM/Ag subjected to uniaxial displacements through an experimental and numeric point of view. Experimental showed that damage proceeds in two ways. First, the formation of a grid crack pattern begins at the ITO layer, and second, the delamination in the ITO/P3HT:PCBM interface. The numerical model analyzed the force and displacements and the absorption/dissipation of strain energy on layers and interfaces of the device. The comparison of the global Young's module for experimental and numeric studies validated the numeric analysis, with results of 4.16 ± 0.05 GPa for experimental and 4.36 ± 0.15 GPa for numeric. Additionally, the model associates the ITO layer with the highest strain energy dissipation or the most prone to failure, which agrees with the experiments. Then, the model successfully predicts the mechanical behavior of OFSC and represents a valuable tool for studying flexible devices and predicting the appearance of mechanical damage when subjected to uniaxial deformations, even being able to avoid potential damage changing parameters such as the thickness of the layers.

Generation of precision preclinical cancer models using regulated in vivo base editing.

Although single-nucleotide variants (SNVs) make up the majority of cancer-associated genetic changes and have been comprehensively catalogued, little is known about their impact on tumor initiation and progression. To enable the functional interrogation of cancer-associated SNVs, we developed a mouse system for temporal and regulatable in vivo base editing. The inducible base editing (iBE) mouse carries a single expression-optimized cytosine base editor transgene under the control of a tetracycline response element and enables robust, doxycycline-dependent expression across a broad range of tissues in vivo. Combined with plasmid-based or synthetic guide RNAs, iBE drives efficient engineering of individual or multiple SNVs in intestinal, lung and pancreatic organoids. Temporal regulation of base editor activity allows controlled sequential genome editing ex vivo and in vivo, and delivery of sgRNAs directly to target tissues facilitates generation of in situ preclinical cancer models.

Best Practice Strategies for Retaining Faculty During Times of Rapid Change.

Faculty retention is an issue of concern to schools and colleges of pharmacy. The reasons why faculty leave are multifactorial but often involve a breach of unwritten contract obligations between the faculty member and the organization. This article provides strategies for retaining faculty based on published literature that include perceived breaches of unwritten contracts and our own perspectives as departmental and university administrators and senior faculty members who have been involved in devising and implementing institutional change. Retention begins with recruitment but then needs to be nurtured during onboarding and as part of the overall enculturation process for new faculty members. Particular attention to the factors that influence the retention of underrepresented minorities must be incorporated to help ensure that pharmacy educators reflect the diversity of the US population.

Potential benefits of microalgae intake against metabolic diseases: beyond spirulina-a systematic review of animal studies.

CONTEXT: Microalgae are a diverse source of bioactive molecules, such as polyphenols, carotenoids, and omega-3 fatty acids, with beneficial properties in biomarkers of metabolic diseases. Unlike the rest of the microalgae genera, Arthrospira sp., commonly called spirulina, has been widely studied. OBJECTIVE: This review aims to describe the current knowledge about microalgae, besides spirulina, focusing on their beneficial properties against metabolic diseases. DATA SOURCES: A systematic research of MEDLINE (via PubMed), Cochrane, and Scopus databases was conducted to identify relevant studies published after January 2012. In vivo animal studies including microalgae consumption, except for spirulina, that significantly improved altered biomarkers related to metabolic diseases were included. These biomarkers included body weight/composition, glucose metabolism, lipid metabolism, oxidative damage, inflammation markers, and gut microbiota. DATA EXTRACTION: After the literature search and the implementation of inclusion and exclusion criteria, 37 studies were included in the revision out of the 132 results originally obtained after the application of the equation on the different databases. DATA ANALYSIS: Data containing 15 microalgae genera were included reporting on a wide range of beneficial results at different levels, including a decrease in body weight and changes in plasma levels of glucose and lipoproteins due to molecular alterations such as those related to gene expression regulation. The most reported beneficial effects were related to gut microbiota and inflammation followed by lipid and glucose metabolism and body weight/composition. CONCLUSIONS: Microalgae intake improved different altered biomarkers due to metabolic diseases and seem to have potential in the design of enriched foodstuffs or novel nutraceuticals. Nevertheless, to advance to clinical trials, more thorough/detailed studies should be performed on some of the microalgae genera included in this review to collect more information on their molecular mechanisms of action.

Squalene encapsulation by emulsification and freeze-drying process: Effects on bread fortification.

In the present work, squalene (SQ) was encapsulated by a conventional emulsion method using egg white protein nanoparticles (EWPn) as a high molecular weight surfactant, followed by a freeze-drying process to obtain an SQ powder ingredient. EWPn was produced by heat treatment at 85°C, 10 min, and pH 10.5. EWPn showed higher emulsifying activity regarding native egg white protein (EWP), highlighting their potential to be used for the SQ encapsulation by an emulsification process. First, we explored the encapsulation conditions using pure corn oil as an SQ carrier. Conditions were oil fraction (0.1-0.2), protein amount (2-5 wt.%), homogenization pressure (100 and 200 bar), and maltodextrin amount (10-20 wt.%). At 0.15 oil fraction, 5 wt.%. protein concentration, 200 bar homogenization pressure, and 20% maltodextrin, the highest encapsulation efficiency (EE) was reached. Then, according to these conditions, SQ was encapsulated to obtain a freeze-dried powder ingredient for bread formulation. The total and free oil of SQ freeze-dried powder were 24.4% ± 0.6% and 2.6% ± 0.1%, respectively, resulting in an EE value of 89.5% ± 0.5%. The physical, textural, and sensory properties of functional bread were not affected by the addition of 5.0% SQ freeze-dried powder. Finally, the bread loaves showed higher SQ stability than the one formulated with unencapsulated SQ. Hence, the encapsulation system developed was suitable for obtaining functional bread based on SQ fortification.

Whole-mount immunofluorescence imaging and isolation of mesothelium-bound immune cell aggregates during mouse peritoneal inflammation.

Resident peritoneal macrophages (resMØs) are crucial for repairing peritoneal injuries and controlling infections by forming mesothelium-bound resMØ-aggregates in the peritoneal wall and omentum. Here we present a protocol to analyze these structures in mouse models of peritoneal inflammation. We describe the dissection, fixation, immunofluorescent staining, and mounting of whole peritoneal wall and omentum samples and subsequent confocal microscopy imaging of resMØ-aggregates. We also detail the steps to isolate resMØ-aggregates for additional studies, including flow cytometry and electron-microscopy-based analysis. For complete details on the use and execution of this protocol, please refer to Vega-Pérez et al. (2021).1.

Mouse Tissue-Resident Peritoneal Macrophages in Homeostasis, Repair, Infection, and Tumor Metastasis.

Large peritoneal macrophages (LPMs) are long-lived, tissue-resident macrophages, formed during embryonic life, developmentally and functionally confined to the peritoneal cavity. LPMs provide the first line of defense against life-threatening pathologies of the peritoneal cavity, such as abdominal sepsis, peritoneal metastatic tumor growth, or peritoneal injuries caused by trauma, or abdominal surgery. Apart from their primary phagocytic function, reminiscent of primitive defense mechanisms sustained by coelomocytes in the coelomic cavity of invertebrates, LPMs fulfill an essential homeostatic function by achieving an efficient clearance of apoptotic, that is crucial for the maintenance of self-tolerance. Research performed over the last few years, in mice, has unveiled the mechanisms by which LPMs fulfill a crucial role in repairing peritoneal injuries and controlling microbial and parasitic infections, reflecting that the GATA6-driven LPM transcriptional program can be modulated by extracellular signals associated with pathological conditions. In contrast, recent experimental evidence supports that peritoneal tumors can subvert LPM metabolism and function, leading to the acquisition of a tumor-promoting potential. The remarkable functional plasticity of LPMs can be nevertheless exploited to revert tumor-induced LPM protumor potential, providing the basis for the development of novel immunotherapeutic approaches against peritoneal tumor metastasis based on macrophage reprogramming.

PEGylation of genistein-loaded bovine serum albumin nanoparticles and its effect on in vitro cell viability and genotoxicity properties.

Self-assembled bovine serum albumin nanoparticles loaded with the isoflavone genistein have shown apoptosis-mediated cytotoxicity against murine mammary adenocarcinoma F3II cells. Due to their protein nature and small particle size (13-15 nm), their parenteral administration could be affected by possible immunogenic reactions and rapid clearance from the bloodstream. To avoid these problems, PEGylation of the systems was achieved in this work by using a 30 kDa methoxy-polyethylene glycol carbonyl imidazole derivative through the reaction between the carbonyl imidazole group and the amino groups of Lys residues on the protein surface, which was confirmed by a 17% reduction in the available amino groups content measured by the o-phthaldialdehyde method. PEGylated isoforms were obtained, showing an increase of particle size from 13 to 15 nm to around 260 nm, and were purified by SEC-FPLC and characterized by SDS-PAGE, DLS and AFM techniques. The effect of PEGylation on BSAnp-Gen cytotoxicity and genotoxicity against F3II cells was evaluated in vitro by MTT assay, flow cytometry analysis and micronucleus assay. From the results, PEGylation produced an improvement of the biological properties of genistein-loaded nanoparticles in terms of cytotoxicity (lower IC50), not affecting the induction of apoptosis, decreasing the genotoxicity of the systems (less induction of micronucleus formation).