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Background: Macrophage activation-like syndrome (MALS) and complex immune dysregulation (CID) often underlie acute respiratory distress (ARDS) in COVID-19. We aimed to investigate the effect of personalized immunotherapy on clinical improvement of critical COVID-19. Methods: In this open-label prospective trial, 102 patients with ARDS by SARS-CoV-2 were screened for MALS (ferritin >4,420 ng/mL) and CID (ferritin ≤4,420 ng/mL and low human leukocyte antigen (HLA)-DR expression on CD14-monocytes). Patients with MALS or CID with increased aminotransferases received intravenous anakinra; those with CID and normal aminotransferases received tocilizumab. The primary outcome was ≥25% decrease in the Sequential Organ Failure Assessment (SOFA) score and/or 50% increase in the respiratory ratio by day 8; 28-day mortality, change of SOFA score by day 28, serum biomarkers, and cytokine production by mononuclear cells were secondary endpoints. Results: The primary study endpoint was met in 58.3% of anakinra-treated patients and in 33.3% of tocilizumab-treated patients (p: 0.01). Most patients in both groups received dexamethasone as standard of care. No differences were found in secondary outcomes, mortality, and SOFA score changes. Ferritin decreased among anakinra-treated patients; interleukin-6, soluble urokinase plasminogen activator receptor, and HLA-DR expression increased among tocilizumab-treated patients. Survivors by day 28 who received anakinra were distributed to lower severity levels of the WHO clinical progression scale. Greater incidence of secondary infections was found with tocilizumab treatment. Conclusion: Immune assessment resulted in favorable anakinra responses among critically ill patients with COVID-19 and features of MALS.

BACKGROUND: Opportunity to redefine the care journeys for those living with primary biliary cholangitis (PBC) includes facilitating access to enhanced (PBC-dedicated) programmes by nonspecialist risk 'flagging' of patients. OBJECTIVE: To develop a nonexpert PBC stratification tool to help care pathway choices (standard vs. enhanced) choices in PBC. METHODS: We included ursodeoxycholic acid-treated patients with PBC from the Global PBC Study Group. The performance of baseline and 1-year clinical markers with transplant-free survival was assessed to develop the 'ABA' tool using Age (A), Bilirubin (B), and Alkaline phosphatase (A). Added value of fibrosis estimation was assessed. RESULTS: 'ABA' classification mapped three risk groups (n = 2226): low [Age > 50 years, bilirubin ≤ 1 × ULN, alkaline phosphatase (ALP) ≤ 3 × ULN], high (Age ≤ 50 years, bilirubin > 1 × ULN, ALP > 3 × ULN), and intermediate (other). Transplant-free survival at 10 years in the low-, intermediate-, and high-risk groups were 89, 77, and 59% at baseline and 86, 76, and 40% at 1 year, respectively. We propose that high-risk patients at baseline be directly triaged to enhanced (PBC-dedicated) care and the remaining be reassessed at 1 year. Modelling showed after 1 year 46% patients were proposed to enhanced care and 54% to standard care. The 'ABA' mapped pathways facilitated identification of patients at risk based on a young age, as compared to traditional liver biochemical stratification. In patients proposed to standard care, estimated fibrosis stage had ongoing prognostic value. CONCLUSION: Nonspecialist use of the 'ABA' risk tool could prioritize care journey choices for patients with PBC.

Proprotein convertase subtilisin/kexin type 9 (Pcsk9) binds to hepatic low-density lipoprotein receptor (LDLR) and induces its internalization and degradation. Pcsk9 inhibition increases LDLR expression by hepatocytes, which causes increased uptake of circulating LDL, thereby reducing plasma LDL-cholesterol. However, by increasing the uptake of LDL by the liver, Pcsk9 inhibition increases the exposure of the liver to cholesterol, which may result in higher risk of steatohepatitis and ever carcinogenesis. We compared Pcsk9-/- knockout (KO) mice and appropriate wild-type (WT) controls of the same strain assigned to a high-fat (15%, wt/wt) diet for 9 months supplemented with 0.25%, 0.5%, or 0.75% dietary cholesterol. Pcsk9 KO mice on a high-fat, high-cholesterol diet exhibited higher levels of hepatic free cholesterol loading and hepatic cholesterol crystallization than their WT counterparts. Pcsk9 KO mice developed crown-like structures of macrophages surrounding cholesterol crystal-containing lipid droplets and hepatocytes, exhibited higher levels of apoptosis, and developed significantly more hepatic inflammation and fibrosis consistent with fibrosing steatohepatitis, including 5-fold and 11-fold more fibrosis at 0.5% and 0.75% dietary cholesterol, respectively. When injected with diethylnitrosamine, a hepatic carcinogen, early-in-life Pcsk9 KO mice were more likely to develop liver cancer than WT mice. Conclusion: Pcsk9 KO mice on high-cholesterol diets developed increased hepatic free cholesterol and cholesterol crystals and fibrosing steatohepatitis with a higher predisposition to liver cancer compared with WT mice. Future studies should evaluate whether patients on long-term treatment with anti-PSCK9 monoclonal antibodies are at increased risk of hepatic steatosis, steatohepatitis or liver cancer, while accounting for concurrent use of statins.

Mango (Mangifera indica L) production in Kenya directly supports approximately 200,000 farmers and many other beneficiaries. Despite this, its production suffers from post-harvest losses due to the fruits' short shelf life in ambient conditions. Calcium maintains cell integrity, strengthens the cell wall, membrane structure, and thus increases shelf life. A completely randomized block design with a split plot arrangement was used to compare the effect of spraying and immersion of ‘Van Dyke' mango fruits at maturity or 15 days later in calcium chloride at different concentrations (0.5%, 1%, 1.5%, or 0%) and times on the fruit ripening rate and organoleptic acceptance. The peel firmness (N), total soluble solids (0Brix), flesh color (Ho), beta carotene (mg/100ml), and carbon dioxide evolution (ml/kg/hr) of fruits were determined at time 0 and every two days for up to eight days in ambient conditions. Additionally, organoleptic characteristics, flesh firmness, calcium concentration (g/mg), and their correlations were determined. Fruits immersed in calcium chloride at maturity had higher retained peel firmness (10.6 N, 10.3 N), deeper flesh color (37.45, 36.78), lower total soluble solids (14, 13.8), a lower carbon dioxide evolution (30.7 ml/kg/hr), higher beta carotene and higher flesh calcium concentration than fruits exposed to other treatments. Fruits sprayed at maturity outperformed those sprayed 15 days later in the studied parameters. Flesh calcium content correlated positively with flesh firmness (r= 0.913, r= 0.852), flesh color (r= 0.828, r= 0.841), fruit aroma (r=0.8199, r=0.841), and negatively with skin shriveling (r=-0.778, r=-0.806) and fruit flavor (r=-0.811, r=-0.829). Flesh firmness correlated negatively with skin shriveling (r=-0.868, r=-0.788) and fruit flavor (r=-0.8869, r=-0.821), but positively with peel color (r=0.9115, r=0.856) and aroma (r=0.907, r=0.848). Skin shriveling was found to have a negative relationship with peel color (r=-0.944, r=-0.93) and aroma (r=-0.944, r=-0.938), but a positive relationship with fruit flavor (r=0.933, r=0.947). Peel color correlated positively with aroma (r=0.979, r=0.977) and negatively with fruit flavor (r=-0.962, r=-0.950), respectively. Despite the effectiveness of post-harvest calcium chloride immersion in extending fruit shelf life, optimal use is advised to avoid deteriorated pulp flavor and increased shriveling. More research is needed to determine how calcium chloride can be made available to the fruit while it is still attached to the tree.

Lampworking is a valuable skill for students in Chemistry and others in science, technology, engineering, and math (STEM) related fields. The activity described here provides instructors the resources to both learn and teach the fundamentals of lampworking. Soft glass, which can be shaped and fused with a Bunsen burner, is used to improve the accessibility of this activity. The Bunsen burners have been modified so that the angle of the flame is between 45 and 60° rather than 90° relative to the base. This modification significantly improves the lampworking experience according to student feedback. Students ranging from Freshmen to Seniors have participated in this activity. This activity introduces students to skills that can be later used with borosilicate glass for scientific glassblowing projects. This activity is both educational and fun for students in STEM fields as well as nonscience majors. Furthermore, this is an excellent professional development opportunity for educators.

Quinolinic acid (QA) is a key intermediate of nicotinic acid (Niacin) which is an essential human nutrient and widely used in food and pharmaceutical industries. In this study, a quinolinic acid producer was constructed by employing comprehensive engineering strategies. Firstly, the quinolinic acid production was improved by deactivation of NadC (to block the consumption pathway), NadR (to eliminate the repression of L-aspartate oxidase and quinolinate synthase), and PtsG (to slow the glucose utilization rate and achieve a more balanced metabolism, and also to increase the availability of the precursor phosphoenolpyruvate). Further modifications to enhance quinolinic acid production were investigated by increasing the oxaloacetate pool through overproduction of phosphoenolpyruvate carboxylase and deactivation of acetate-producing pathway enzymes. Moreover, quinolinic acid production was accelerated by assembling NadB and NadA as an enzyme complex with the help of peptide-peptide interaction peptides RIAD and RIDD, which resulted in up to 3.7 g/L quinolinic acid being produced from 40 g/L glucose in shake-flask cultures. A quinolinic acid producer was constructed in this study, and these results lay a foundation for further engineering of microbial cell factories to efficiently produce quinolinic acid and subsequently convert this product to nicotinic acid for industrial applications.

Prenylated indole alkaloids featuring spirooxindole rings possess a 3R or 3S carbon stereocenter, which determines the bioactivities of these compounds. Despite the stereoselective advantages of spirooxindole biosynthesis compared with those of organic synthesis, the biocatalytic mechanism for controlling the 3R or 3S-spirooxindole formation has been elusive. Here, we report an oxygenase/semipinacolase CtdE that specifies the 3S-spirooxindole construction in the biosynthesis of 21R-citrinadin A. High-resolution X-ray crystal structures of CtdE with the substrate and cofactor, together with site-directed mutagenesis and computational studies, illustrate the catalytic mechanisms for the possible β-face epoxidation followed by a regioselective collapse of the epoxide intermediate, which triggers semipinacol rearrangement to form the 3S-spirooxindole. Comparing CtdE with PhqK, which catalyzes the formation of the 3R-spirooxindole, we reveal an evolutionary branch of CtdE in specific 3S spirocyclization. Our study provides deeper insights into the stereoselective catalytic machinery, which is important for the biocatalysis design to synthesize spirooxindole pharmaceuticals., Prenylated indole alkaloids contain spirooxindole rings with a 3R or 3S carbon stereocenter, which determines their bioactivities, but the biocatalytic mechanism controlling the 3R- or 3S-spirooxindole formation was unclear. Here, the authors report the biochemical and structural characterization of the oxygenase/semipinacolase CtdE that catalyses the 3S-spirooxindole construction in the biosynthesis of 21R-citrinadin A.

Palladium-catalyzed cross-coupling protocols have become a cornerstone in organic synthesis. Here, a gas-phase model of the Hiyama cross-coupling reaction was designed to shed light on the roles of coordinated anions (fluoride versus chloride) in transmetalation from Si to Pd. A combination of mass spectrometry experiments and DFT calculations was used. The ligated palladium fluoride and chloride cationic complexes, [(phen)Pd(X)]+ (X = F and Cl), readily react with vinyltrimethylsilane, Me3Si(CH═CH2), via transmetalation to give [(phen)Pd(CH═CH2)]+ as the major product. DFT calculations reveal that this transmetalation reaction is concerted and proceeds via a four-centered transition state, illustrating the role of coordinated halide in this gas-phase system. Two minor side products are observed corresponding to transmetalation to give [(phen)Pd(CH3)]+ and [(phen)Pd(SiMe2X)]+. DFT calculations suggest that these arise from the same initial Si to Pd methyl transmetalation pathway to give the [(phen)Pd(CH3) + Me2(CH═CH2)SiX]+ intermediate, which either then loses Me2(CH═CH2)SiX or reacts via C–C bond coupling to ultimately yield propene and [(phen)Pd(SiMe2X)]+. [(phen)Pd(CH═CH2)]+ undergoes a reaction with a second molecule of vinyltrimethylsilane to form an adduct, which upon collision-induced dissociation liberates 1,3-butadiene to form [(phen)Pd(SiMe3)]+. DFT calculations suggest a mechanism in which C–C bond formation is followed by migration of SiMe3 from C to Pd. Links between the observed gas-phase chemistry and solution-phase Pd-mediated homocoupling reactions of vinyltrimethylsilanes are explored.

Aims Infection by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) may lead to the development of severe respiratory failure. In hospitalized-patients, prompt interruption of the virus-driven inflammatory process by using combination treatments seems theoretically of outmost importance. Our aim was to investigate the hypothesis of multifaceted management of these patients. Methods A treatment algorithm based on ferritin was applied in 311 patients (67.2% males; median age 63-years; moderate disease, n=101; severe, n=210). Patients with ferritin

Statistical analysis is performed on the totalionizing-dose (TID) response of fourth-generation silicon–germanium heterojunction bipolar transistors, revealing an increasing variance in base current with increasing dose. A 10-fold increase is observed after 2 Mrad(SiO2) of radiation exposure. The postirradiation performance is shown to be poorly correlated with initial performance, revealing potential difficulties for radiation hardness assurance testing. Simple analytical theory is used to demonstrate how TID exacerbates performance variations due to preexisting process-induced variations. Then, technology computer-aided design (TCAD) simulations are used to investigate the fundamental mechanisms driving these statistical changes, particularly process variations associated with the doping profiles. Simulation results reveal potential methods to both predict TID response and mitigate it at the process level. Finally, implications for hardness assurance testing and radiation-tolerant device and circuit design are discussed, especially the need to consider postirradiation variability in the design process for radiation-intense applications.

Lipoprotein disorders are a major risk factor for atherosclerotic neuro-cardiovascular disease (ACVD) and are heavily influenced by lifestyle, including alcohol drinking. Moderate drinkers have a lower ACVD risk than abstainers due to their higher levels of high-density lipoprotein (HDL) cholesterol, an important protective factor against ACVD. On the contrary, heavy drinking increases ACVD risk. According to an extensive literature body, ethanol intoxication modifies lipid serum profile and induces endothelial dysfunction. Single nucleotide polymorphisms may influence the relationship between alcohol drinking, HDL cholesterol level, and atherosclerotic risk. The risk of ACVD in heavy drinkers seems enhanced in patients with apolipoprotein E4 allele, interleukin- 6-174 polymorphism, and cholesteryl ester transfer protein TaqIB polymorphism. Apolipoprotein E4 is a known risk factor for ACVD, while apolipoprotein E2 has mixed effects. Therefore, even if a “protective role” may be attributed to moderate drinking, this effect cannot be extended to everyone.

Selective reduction of CO2 to formate represents an ongoing challenge in photoelectrocatalysis. To provide mechanistic insights, we investigate the kinetics of hydride transfer (HT) from a series o...

Fluorescent nucleobase surrogates capable of Watson–Crick hydrogen bonding are essential probes of nucleic acid structure and dynamics, but their limited brightness and short absorption and emission wavelengths have rendered them unsuitable for single-molecule detection. Aiming to improve on these properties, we designed a new tricyclic pyrimidine nucleoside analogue with a push–pull conjugated system and synthesized it in seven sequential steps. The resulting C-linked 8-(diethylamino)benzo[b][1,8]naphthyridin-2(1H)-one nucleoside, which we name ABN, exhibits ε442 = 20 000 M−1 cm−1 and Φem,540 = 0.39 in water, increasing to Φem = 0.50–0.53 when base paired with adenine in duplex DNA oligonucleotides. Single-molecule fluorescence measurements of ABN using both one-photon and two-photon excitation demonstrate its excellent photostability and indicate that the nucleoside is present to > 95% in a bright state with count rates of at least 15 kHz per molecule. This new fluorescent nucleobase analogue, which, in duplex DNA, is the brightest and most red-shifted known, is the first to offer robust and accessible single-molecule fluorescence detection capabilities., Fluorescent nucleoside analogue ABN is readily detected at the single-molecule level and retains a quantum yield >50% in duplex DNA oligonucleotides.

Protein Kinase C (PKC) is a member of the AGC subfamily of kinases and regulates a wide array of signaling pathways and physiological processes. Protein–protein interactions involving PKC and its scaffolding partners dictate the spatiotemporal dynamics of PKC activity, including its access to activating second messenger molecules and potential substrates. While the A Kinase Anchoring Protein (AKAP) family of scaffold proteins universally bind PKA, several were also found to scaffold PKC, thereby serving to tune its catalytic output. Targeting these scaffolding interactions can further shed light on the effect of subcellular compartmentalization on PKC signaling. Here we report the development of two hydrocarbon stapled peptides, CSTAD5 and CSTAD6, that are cell permeable and bind PKC to disrupt PKC–gravin complex formation in cells. Both constrained peptides downregulate PMA-induced cytoskeletal remodeling that is mediated by the PKC–gravin complex as measured by cell rounding. Further, these peptides downregulate PKC substrate phosphorylation and cell motility. To the best of our knowledge, no PKC-selective AKAP disruptors have previously been reported and thus CSTAD5 and CSTAD6 are novel disruptors of PKC scaffolding by AKAPs and may serve as powerful tools for dissecting AKAP-localized PKC signaling., We report the development of AKAP derived, conformationally constrained peptides designed to probe AKAP-localized PKC. The lead peptides, CSTAD5 and CSTAD6 permeate cells, bind PKC, disrupt its scaffolding by AKAPs to inhibit its scaffolded activity.

Natural products and natural product-derived compounds have been widely used for pharmaceuticals for many years, and the search for new natural products that may have interesting activity is ongoing. Abyssomicins are natural product molecules that have antibiotic activity via inhibition of the folate synthesis pathway in microbiota. These compounds also appear to undergo a required [4 + 2] cycloaddition in their biosynthetic pathway. Here we report the structure of an flavin adenine dinucleotide-dependent reductase, AbsH3, from the biosynthetic gene cluster of novel abyssomicins found in Streptomyces sp. LC-6-2.

Nanoparticles (NPs) produced from amphiphilic derivatives of poly-N-vinylpyrrolidone (Amph-PVP), composed of various molecular weight polymeric hydrophilic fragments linked into hydrophobic n-alkyl chains of varying lengths, were previously shown to exert excellent biocompatibility. Although routes of administration can be different, finally, most nanosystems enter the blood circulation or lymphatic vessels, and by this, they establish direct contact with endothelial cells. In this study, Amph-PVP NPs and fluorescently labeled Amph-PVP-based NPs, namely "PVP" NPs (Amph-PVP-NPs (6000 Da) unloaded) and "F"-NPs (Amph-PVP-NPs (6000 Da) loaded with fluorescent FITC), were synthesized to study Amph-PVP NPs interactions with HMEC-1 endothelial cells. PVP NPs were readily uptaken by HMEC-1 cells in a concentration-dependent manner, as demonstrated by immunofluorescence imaging. Upon uptake, the FITC dye was localized to the perinuclear region and cytoplasm of treated cells. The generation of lipopolysaccharide (LPS)-induced activated endothelium model revealed an increased uptake of PVPNPs, as shown by confocal microscopy. Both unloaded PVP NPs and F-NPs did not affect EC viability in the 0.01 to 0.066 mg/mL range. Furthermore, we focused on the potential immunological activation of HMEC-1 endothelial cells upon PVPNPs treatment by assessing the expression of their E-Selectin, ICAM-1, and VCAM-1 adhesion receptors. None of the adhesion molecules were affected by NP treatments of both activated by LPS and nonactivated HMEC-1 cells, at the utilized concentrations (p = NS). In this study, PVP (6000 Da) NPs were used to encapsulate indomethacin, a widely used anti-inflammatory drug. The synthesized drug carrier complex did not affect HMEC-1 cell growth and expression of E-selectin, ICAM-1, and VCAM-1 adhesion receptors. In summary, PVP-based NPs are safe for use on both basal and activated endothelium, which more accurately mimics pathological conditions. Amph-PVP NPs are a promising drug delivery system.

Low-noise amplifiers (LNAs) are necessary components for any communications system. Single-event transients (SETs) induced by energetic particles in space can corrupt the data in an RF receiver. Using p-n-p silicon–germanium (SiGe) heterojunction bipolar transistors (HBTs), instead of their more common n-p-n counterparts, has been shown to be an effective mitigation technique in digital, analog, and RF circuits. Since p-n-p SiGe HBTs tend to have lower performance than n-p-n SiGe HBTs, in this article, the tradeoffs between performance and SET robustness have been evaluated through the use of simulations. Two LNAs were designed using an algorithmic design technique: one using only n-p-n SiGe HBTs, and the other using only p-n-p devices. The n-p-n LNA had higher peak gain and lower noise figure at the center frequency of 5 GHz when compared to the p-n-p LNA, by 3.2 and 0.4 dB, respectively. However, the n-p-n LNA also produced transients with amplitudes larger than the p-n-p LNA across all simulated linear energy transfers. Although the p-n-p LNA has lower overall performance, it is also more robust to SETs. Thus, the choice between using n-p-n and p-n-p SiGe HBTs for an RF design will depend on application requirements.

The crystallization behavior of amorphous barium titanate (BaTiO3) thin films was studied as a function of annealing temperature from 500 to 1000 °C. Quantitative phase analysis by grazing incidence X-ray diffractometry revealed that the metastable hexagonal phase preferentially nucleates at lower temperatures with a transition to stable cubic nuclei at higher temperatures. The predominance of the metastable hexagonal-phase nucleation at lower temperatures suggests that it has the lowest nucleation barrier, in accordance with the Ostwald’s step rule. To help induce the nucleation of the cubic phase at lower temperatures, we explored the effects of electric fields on the crystallization behavior and found that direct-current fields moderately enhanced the cubic phase fraction in the 500–700 °C temperature range. Although the nucleation barrier of the cubic phase in the presence of an electric should be lowered more significantly with respect to that of the hexagonal phase because of the former’s higher relative permittivity, the effect arising from the change in volume free energy should only become significant at electric field strengths in the range of MV cm−1, an order of magnitude higher than the experimentally accessible fields in the present study. This suggests alternative, perhaps interface-mediated, mechanisms by which the electric field modifies the nucleation behavior of BaTiO3.

Cement, the most used building material amounts to an annual world production of 3.5 Gt. Sulfobelite cements, SB, require lower kiln temperature and emit less CO2, compared to Ordinary Portland cement, OPC. The main difference of SB from OPC is the reduction of alite (CaO)3SiO2, the increase of belite (CaO)2SiO2 and the formation of yeelimite (CaO)4(Al2O3)3SO3, also called “Klein’s salt”. Industrial byproducts, such as Flue Gas Desulphurization gypsum, FGD and dry Red Mud, the so–called bauxite residue, BR can also be used as raw materials, decreasing the CO2 footprint. In the present study, SB cements were prepared in laboratory scale from FGD and BR, as well as, conventional raw materials. All compositions were characterized by XRD, Q–XRD, optical microscopy, SEM and were tested for their compressive strength, soundness, coagulation, etc. The main result of this work is that the studied industrial byproducts, FGD and BR, can be used as alternative raw materials for the raw meals of SB cements, accomplishing low–temperature clinkering at 1330 °C and lower quantities of conventional raw materials. Conclusions suggest that SB cements can be a promising eco–friendly building material and can be used in other applications such as 3-D printed building, rapid-setting mortars, rapid sealants, etc. Moreover, this is an attempt to apply circular economy in cement production utilizing the established production route of cement industries.

Previously we reported that site-specific modification of the human granulocyte-macrophage colony-stimulating factor (GM-CSF) A3C analog with polyethylene glycol (PEG) dramatically improved the pharmacokinetic properties of the protein in rats. However, we could not evaluate the hematological properties of the PEG-A3C protein in rats because human GM-CSF is inactive in rodents. To study the biological effects of PEGylated GM-CSF analogs in rodents we created a homologous site-specific PEGylated murine (mu) GM-CSF (T3C) protein. muGM-CSF and the T3C protein were expressed in Escherichia coli and purified by column chromatography. The purified T3C protein was covalently modified with a linear 20 kDa- or a branched 40 kDa-maleimide-PEG, and the monoPEGylated proteins purified by column chromatography. muGM-CSF, T3C and the two PEG-T3C proteins had comparable in vitro biological activities, as measured by stimulation of proliferation of the murine FDC-P1 cell line. The PEG-T3C proteins had 10- to 25-fold longer circulating half-lives than muGM-CSF and stimulated greater and longer lasting increases in neutrophils and white blood cells than muGM-CSF following a single intravenous or subcutaneous administration to rats. Treatment of rats made neutropenic with cyclophosphamide with the PEG-T3C proteins shortened the time for recovery of neutrophils to normal levels from 9 or 10 days to 5 or 6 days, whereas muGM-CSF showed no benefit versus vehicle solution. Acceleration of neutrophil recovery in cyclophosphamide-treated rats required a minimum of three PEG-T3C treatments over five days. The PEG-T3C proteins should prove useful for evaluating the potential therapeutic benefits of GM-CSF and long-acting GM-CSF proteins in rodent disease models.

Hemoglobin functions as a tetrameric oxygen transport protein, with each subunit containing a heme cofactor. Its denaturation, either in vivo or in vitro, involves autoxidation to methemoglobin, followed by cofactor loss and globin unfolding. We have proposed a global disassembly scheme for human methemoglobin, linking hemin (ferric protoporphyrin IX) disassociation and apoprotein unfolding pathways. The model is based on the evaluation of circular dichroism and visible absorbance measurements of guanidine-hydrochloride-induced disassembly of methemoglobin and previous measurements of apohemoglobin unfolding. The populations of holointermediates and equilibrium disassembly parameters were estimated quantitatively for adult and fetal hemoglobins. The key stages are characterized by hexacoordinated hemichrome intermediates, which are important for preventing hemin disassociation from partially unfolded, molten globular species during early disassembly and late-stage assembly events. Both unfolding experiments and independent small angle x-ray scattering measurements demonstrate that heme disassociation leads to the loss of tetrameric structural integrity. Our model predicts that after autoxidation, dimeric and monomeric hemichrome intermediates occur along the disassembly pathway inside red cells, where the hemoglobin concentration is very high. This prediction suggests why misassembled hemoglobins often get trapped as hemichromes that accumulate into insoluble Heinz bodies in the red cells of patients with unstable hemoglobinopathies. These Heinz bodies become deposited on the cell membranes and can lead to hemolysis. Alternatively, when acellular hemoglobin is diluted into blood plasma after red cell lysis, the disassembly pathway appears to be dominated by early hemin disassociation events, which leads to the generation of higher fractions of unfolded apo subunits and free hemin, which are known to damage the integrity of blood vessel walls. Thus, our model provides explanations of the pathophysiology of hemoglobinopathies and other disease states associated with unstable globins and red cell lysis and also insights into the factors governing hemoglobin assembly during erythropoiesis.

It is of great economic interest to produce succinate from low-grade carbon sources, e.g., lignocellulosic biomass hydrolysate, which mainly contains glucose and xylose. Inactivation of the glucose uptake system PtsG was evaluated for succinate production from xylose-rich feedstocks. Strains with integration of succinate production modules into the chromosome of Escherichia coli were then constructed. These strains have better succinate production performance from xylose-rich feedstocks than strain FZ560 harboring pHL413KF1. Glucose utilization was enhanced in FZ661T by manipulation of the gal operon to allow efficient use of the high-concentration glucose in woody biomass hydrolysate. Up to 906.7 mM (107.0 g/L) succinate was produced from mixed sugars in fed-batch fermentation and more than 461.7 mM (54.5 g/L) succinate was produced from woody hydrolysate in a batch fermentation. In this study, FZ661T was able to produce succinate from woody hydrolysate in minimal medium efficiently, making it attractive for industrial applications in succinate production.

The present study concerns with an optimization method of Solid Oxide Fuel Cell (SOFC) systems for electricity generation, directly fed by CH4 (methane) or C2H5OH (ethanol). By considering a detailed thermodynamic analysis, an innovative simulation model, namely THERMAS (THERmodynamic MAthematical Simulation), was designed and implemented. A specific SOFC-based system was selected to be the reference simulation, which is assumed to be equipped with heat exchangers (preheaters), a reformer, a SOFC-stack system and an afterburner. THERMAS allows for an extended parametric analysis in terms of energy and exergy and offers the opportunity to investigate all the operational characteristics that affect system's efficiency. The optimization process relies on the difference between the energy and exergy efficiency, where an OPF (OPtimization Factor) has been introduced and particularly estimated for each simulated scenario, based on several operational parameters, such as fuel composition, extension of chemical reactions and temperatures. The introduction of this OPF is actually an innovative improvement towards an easy-to-use optimization of SOFC systems. It is found that the design of a SOFC-based power plant fueled by pure hydrocarbons, has to be carefully simulated before its, otherwise the purification of fuels sounds meaningless.

It is of great economic interest to produce succinate from low-grade carbon sources, which can make it more economically competitive against petrochemical-based succinate. Galactose sugars constitute a significant fraction of the soluble carbohydrate in a meal from agricultural sources which is considered a low value or waste byproduct of oilseed processing. To improve the galactose utilization, the effect of galR and glk on sugars uptake was investigated by deactivation of each gene in three previously engineered host strains. As expected, glk plays an important role in glucose uptake, while, the effect of deactivation of galR is highly dependent on the strength of the downstream module (succinate production module). A new succinate producer FZ661T was constructed by enhancement of the succinate producing module and manipulation of the gal operon. The succinate productivity reached 4.57 g/L/hr when a mixed sugar feedstock was used as a carbon source in shake-flask fermentation, up to 812 mM succinate was accumulated in 80 hr in fed-batch fermentation. When SoyMolaGal hydrolysate was used as a carbon source, 628 mM (74 g/L) succinate was produced within 72 hr. In this study, we demonstrate that FZ661T can produce succinate quickly with relatively high yield, giving it the potential for industrial application.

A novel approach for two-photon absorption (TPA) pulsed-laser testing produces extended charge deposition profiles that are analogous to those produced by heavy ions. In this approach, which utilizes an axicon rather than a spherical lens, the conventional Gaussian beam is replaced by a quasi-Bessel beam. The key feature of a quasi-Bessel beam, relevant to pulsed-laser single-event effects (PL-SEE) studies, is that its radial size ( $\sim 1~\mu \text{m}$ ) remains constant over the axial length of the beam (several 100 s of $\mu \text{m}$ ) resulting in a charge-deposition profile that more closely mimics that produced by a heavy ion. This elongated charge distribution directly lends itself to a simple and intuitive description in terms of linear-energy transfer (LET), facilitating laser/ion correlation studies. Experimental results comparing a conventional TPA PL-SEE focusing geometry and the axicon focusing geometry are presented for three different test vehicles: a 45-nm silicon-on-insulator (SOI) nFET, a bulk silicon photodiode, and an LM124 operational amplifier. By using the axicon approach, strong laser/ion correlation is observed in the silicon photodiode for single-event transients (SET) across a wide range of LETs. The correlation is achieved by “tuning” the laser to the desired LET rather than relying on preexisting heavy-ion data, and therefore, this approach holds promise as a predictive tool.

The propagation of single-event transients from the electrical to the photonic domain in a segmented Mach–Zehnder modulator was investigated using pulsed-laser measurements and lumerical simulations. Although electrical transients can heavily degrade the input data to the modulator, almost all of the degradation can be suppressed once it is converted into the optical domain. The mitigation of transients is primarily accomplished by increasing $V_\pi $ of each segment, making electrical transients up to 6 V essentially negligible. A “folding” effect of the optical transient is also explained in relation to $V_\pi $ . Two radiation-hardening-by-design (RHBD) approaches are suggested to mitigate transient effects.

The effects of 14-MeV neutron displacement damage (DD) on waveguide (WG)-integrated germanium-on-silicon p-i-n photodiodes (PDs) for silicon photonics have been investigated up to the fluences of $7.5\times 10^{12}$ n/cm2 (14 MeV) or $1.4\times 10^{13}\,\,\text{n}_{1\text {-MeV}_{\mathrm{ eq}}}$ /cm2(Si). This article includes the measurements of dark current–voltage characteristics across temperature from 150 to 375 K, measurements of PD junction capacitance, spectral response measurements from 1260 to 1360 nm, and frequency-response measurements. The devices are found to be susceptible to DD-induced carrier removal effects; however, they also continue to operate without meaningful impact to performance for the DD dose levels examined. Since the PD test chips include silicon photonic integrated grating couplers and WGs, which carry the optical signal to the PD, some assessment of the impact of DD on these passive devices can also be inferred. This article does not examine the short-term annealing or transient behavior of the DD, and instead, it has only considered the lasting damage that remains after any initial period of room-temperature annealing.

INTRODUCTION: In primary biliary cholangitis (PBC), bilirubin and alkaline phosphatase (ALP) are widely established as independent predictors of prognosis. Current treatment goals do not aim for normalization of surrogate markers because their association with survival has not been defined. METHODS: The patient cohort from the GLOBAL PBC Study Group was used, comprising of long-term follow-up data from European and North American centers. Ursodeoxycholic acid-treated and untreated patients with bilirubin levels ≤1 × upper limit of normal (ULN) at baseline or 1 year were included. The association of normal ALP with transplant-free survival was assessed in a subgroup with ALP ≤1.67 × ULN at 1 year. Optimal thresholds of bilirubin and ALP to predict liver transplantation (LT) or death were evaluated. RESULTS: There were 2,281 patients included in the time zero cohort and 2,555 patients in the 1-year cohort. The bilirubin threshold with the highest ability to predict LT or death at 1 year was 0.6 × ULN (hazard ratio 2.12, 95% CI 1.69-2.66, P < 0.001). The 10-year survival rates of patients with bilirubin ≤0.6 × ULN and >0.6 × ULN were 91.3% and 79.2%, respectively (P < 0.001). The risk for LT or death was stable below the bilirubin levels of 0.6 × ULN, yet increased beyond this threshold. Ursodeoxycholic acid-induced reduction in bilirubin below this threshold was associated with an 11% improvement in 10-year survival. Furthermore, ALP normalization was optimal, with 10-year survival rates of 93.2% in patients with ALP ≤ 1 × ULN and 86.1% in those with ALP 1.0-1.67 × ULN. DISCUSSION: Attaining bilirubin levels ≤0.6 × ULN or normal ALP are associated with the lowest risk for LT or death in patients with PBC. This has important implications for treatment targets.

A comparison of heavy-ion-induced single-event transients (SETs) in silicon–germanium heterojunction bipolar transistors (SiGe HBTs) fabricated on both bulk and silicon-on-insulator (SOI) substrates is presented. Experimental heavy-ion data show a reduction of sensitive volume of $\approx 350\times $ for the SiGe HBTs fabricated on SOI compared to those on a bulk substrate. Furthermore, the results indicate that the sensitive volume of the SOI SiGe HBTs is confined to the intrinsic device. These results have been confirmed using pulsed laser data and TCAD simulations. Limiting the charge collection depth of SiGe HBTs by fabricating them on SOI facilitates correlation of ion and laser results. The improved ease of correlation is a result of suppressing diffusive charge collection mechanisms that are difficult to replicate with the charge deposition profile generated by two-photon absorption using a tightly focused laser beam. However, a potential issue with pulsed laser testing is determining the loss of the pulse energy through the buried oxide, which requires detailed knowledge of the semiconductor platform. Overall, the reduction of sensitive volume for the SOI SiGe HBTs compared to those on a bulk substrate is a clear advantage from a single-event upset vulnerability perspective.

We report the identification of the ter gene cluster responsible for the formation of the p-terphenyl derivatives terfestatins B and C and echoside B from the Appalachian Streptomyces strain RM-5-8. We characterize the function of TerB/C, catalysts that work together as a dual enzyme system in the biosynthesis of natural terphenyls. TerB acts as a reductase and TerC as a dehydratase to enable the conversion of polyporic acid to a terphenyl triol intermediate. X-ray crystallography of the apo and substrate-bound forms for both enzymes provides additional mechanistic insights. Validation of the TerC structural model via mutagenesis highlights a critical role of arginine 143 and aspartate 173 in catalysis. Cumulatively, this work highlights a set of enzymes acting in harmony to control and direct reactive intermediates and advances fundamental understanding of the previously unresolved early steps in terphenyl biosynthesis.

Macromolecular function commonly involves rapidly reversible alterations in three-dimensional structure (conformation). To allow these essential conformational changes, macromolecules must possess higher order structures that are appropriately balanced between rigidity and flexibility. Because of the low stabilization free energies (marginal stabilities) of macromolecule conformations, temperature changes have strong effects on conformation and, thereby, on function. As is well known for proteins, during evolution, temperature-adaptive changes in sequence foster retention of optimal marginal stability at a species’ normal physiological temperatures. Here, we extend this type of analysis to messenger RNAs (mRNAs), a class of macromolecules for which the stability–lability balance has not been elucidated. We employ in silico methods to determine secondary structures and estimate changes in free energy of folding (ΔG(fold)) for 25 orthologous mRNAs that encode the enzyme cytosolic malate dehydrogenase in marine mollusks with adaptation temperatures spanning an almost 60 °C range. The change in free energy that occurs during formation of the ensemble of mRNA secondary structures is significantly correlated with adaptation temperature: ΔG(fold) values are all negative and their absolute values increase with adaptation temperature. A principal mechanism underlying these adaptations is a significant increase in synonymous guanine + cytosine substitutions with increasing temperature. These findings open up an avenue of exploration in molecular evolution and raise interesting questions about the interaction between temperature-adaptive changes in mRNA sequence and in the proteins they encode.

Background: We aimed to investigate the potential beneficial effect of immunomodulation therapy on the thromboembolic risk in hospitalized COVID-19 patients. Methods: We searched PubMed and Scopus for randomized trials reporting the outcomes of venous thromboembolism (VTE), ischemic stroke or systemic embolism, myocardial infarction, any thromboembolic event, and all-cause mortality in COVID-19 patients treated with immunomodulatory agents. Odds ratios (OR) and 95% confidence intervals (CI) were calculated using the Mantel–Haenszel random effects method. Results: Among 8499 patients hospitalized with COVID-19, 4638 were treated with an immunomodulatory agent, 3861—with usual care only. Among the patients prescribed immunomodulatory agents, there were 1.77 VTEs per 100 patient-months compared to 2.30 among those treated with usual care (OR: 0.84, 95% CI: 0.61–1.16; I2: 0%). Among the patients who received an interleukin 6 (IL-6) antagonist, VTEs were reported in 12 among the 1075 patients compared to 20 among the 848 receiving the usual care (OR: 0.52, 95% CI: 0.22–1.20; I2: 6%). Immunomodulators as an add-on to usual care did not reduce the risk of stroke or systemic embolism (OR: 1.10, 95% CI: 0.50–2.40; I2: 0%) or of myocardial infarction (OR: 1.06, 95% CI: 0.47–2.39; I2: 0%) and there was a nonsignificant reduction in any thromboembolic event (OR: 0.86, 95% CI: 0.65–1.14; I2: 0%). Conclusions: We did not identify a statistically significant effect of immunomodulation on prevention of thromboembolic events in COVID-19. However, given the large effect estimate for VTE prevention, especially in the patients treated with IL-6 antagonists, we cannot exclude a potential effect of immunomodulation.

Marine cyanobacteria are infected by phages whose genomes encode ferredoxin (Fd) electron carriers. These Fds are thought to redirect the energy harvested from light to phage-encoded oxidoreductases that enhance viral fitness, but it is unclear how the biophysical properties and partner specificities of phage Fds relate to those of photosynthetic organisms. Here, results of a bioinformatics analysis using a sequence similarity network revealed that phage Fds are most closely related to cyanobacterial Fds that transfer electrons from photosystems to oxidoreductases involved in nutrient assimilation. Structural analysis of myovirus P-SSM2 Fd (pssm2-Fd), which infects the cyanobacterium Prochlorococcus marinus, revealed high levels of similarity to cyanobacterial Fds (root mean square deviations of ≤0.5 Å). Additionally, pssm2-Fd exhibited a low midpoint reduction potential (–336 mV versus a standard hydrogen electrode), similar to other photosynthetic Fds, although it had lower thermostability (T(m) = 28 °C) than did many other Fds. When expressed in an Escherichia coli strain deficient in sulfite assimilation, pssm2-Fd complemented bacterial growth when coexpressed with a P. marinus sulfite reductase, revealing that pssm2-Fd can transfer electrons to a host protein involved in nutrient assimilation. The high levels of structural similarity with cyanobacterial Fds and reactivity with a host sulfite reductase suggest that phage Fds evolved to transfer electrons to cyanobacterially encoded oxidoreductases.

Marine cyanobacteria are infected by phage whose genomes encode ferredoxin (Fd) electron carriers. While these Fds are thought to redirect the energy harvested from light to phage-encoded oxidoreductases that enhance viral fitness, it is not clear how the biophysical properties and partner specificities of phage Fds relate to those in photosynthetic organisms. Bioinformatic analysis using a sequence similarity network revealed that phage Fds are most closely related to cyanobacterial Fds that transfer electrons from photosystems to oxidoreductases involved in nutrient assimilation. Structural analysis of myovirus P-SSM2 Fd (pssm2-Fd), which infectsProchlorococcus marinus, revealed high similarity to cyanobacterial Fds (≤0.5 Å RMSD). Additionally, pssm2-Fd exhibits a low midpoint reduction potential (−336 mV vs. SHE) similar to other photosynthetic Fds, albeit lower thermostability (Tm= 28°C) than many Fds. When expressed in anEscherichia colistrain with a sulfite assimilation defect, pssm2-Fd complemented growth when coexpressed with aProchlorococcus marinussulfite reductase, revealing that pssm2-Fd can transfer electrons to a host protein involved in nutrient assimilation. The high structural similarity with cyanobacterial Fds and reactivity with a host sulfite reductase suggest that phage Fds evolved to transfer electrons to cyanobacterial-encoded oxidoreductases.